TREATMENT OF INFLAMMATORY DISEASE USING INGESTIBLE DEVICE TO RELEASE IMMUNE MODULATOR

Abstract

This disclosure features methods and compositions for treating inflammatory disorders or conditions that arise in a tissue originating from the endoderm using an immune modulator.

Claims

1.-229. (canceled)

230. A method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm in a subject in need thereof, the method comprising using an ingestible device to administer to the subject a pharmaceutical composition comprising a therapeutically effective amount of an immune modulator, wherein the pharmaceutical composition is released from the ingestible device in the small intestine or large intestine of the subject, thereby delivering the immune modulator to a location in the GI tract of the subject.

231. The method of claim 230, wherein the pharmacodynamic effects from releasing the immune modulator to the small intestine or large intestine of the subject are observed in one or more tissues proximal to the site of release.

232. The method of claim 231, wherein the pharmacodynamic effects are observed in one or more of the mesenteric lymph nodes (MSN) or the organs and tissues that drain into the MSN.

233. The method of claim 230, wherein the release of the pharmaceutical composition to the small intestine or large intestine provides one or more pharmacodynamic effects selected from: (i) suppression of a local inflammatory response and (ii) maintaining the systemic immune response.

234. The method of claim 233, wherein the release of the pharmaceutical composition to the small intestine or large intestine results in (i) a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T cells in a Peyer's patch in the subject, and/or (ii) an increase in the level of T cells in the blood of the subject; each as compared to the corresponding level in a subject administered the same amount of the immune modulator subcutaneously or intravenously.

235. The method of claim 234, wherein the level of T cells in the mesenteric lymph node is the level of Th memory cells in the mesenteric lymph node.

236. The method of claim 234, wherein the level of T cells in the Peyer's patch is the level of Th memory cells in the Peyer's patch.

237. The method of claim 234, wherein the level of T cells in the blood is the level of Th memory cells in the blood.

238. The method of claim 230, wherein the therapeutically effective amount of the immune modulator administered via the ingestible device is less than an amount that is effective when the immune modulator is administered subcutaneously or intravenously.

239. The method of claim 230, wherein the release of the pharmaceutical composition from the ingestible device comprises topical delivery of the immune modulator.

240. The method of claim 230, wherein releasing the pharmaceutical composition from the ingestible device is triggered by one or more of: a pH in the jejunum from 6.1 to 7.2, a pH in the mid small bowel from 7.0 to 7.8, a pH in the ileum from 7.0 to 8.0, a pH in the right colon from 5.7 to 7.0, a pH in the mid colon from 5.7 to 7.4, a pH in the left colon from 6.3 to 7.7, such as 7.0.

241. The method of claim 230, wherein the pharmaceutical composition is released as a bolus.

242. The method of claim 230, wherein the tissue originating from the endoderm is selected from the group consisting of the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder.

243. The method of claim 230, wherein the tissue originating from the endoderm is selected from the group consisting of the liver, the pancreas, the intestines, and the gallbladder.

244. The method of claim 230, wherein the inflammatory disease or condition is selected from the group consisting of gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary sclerosing cholangitis, pancreatitis, interstitial cystitis, asthma, chronic obstructive pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjgren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary sclerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagille syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, intra- and extrahepatic cholestasis, hereditary forms of cholestasis, PFIC1, gall stones, choledocholithiasis, malignancy causing obstruction of the biliary tree, scratching and/or pruritus due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radiation-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis.

245. The method of claim 230, wherein the inflammatory disease or condition is inflammation of the liver.

246. The method of claim 230, wherein the immune modulator is an integrin inhibitor.

247. The method of claim 246, wherein the integrin inhibitor is selected from the group consisting of an inhibitory nucleic acid, a fusion protein, an integrin antagonist, a cyclic peptide, a disintegrin, a peptidomimetic, a small molecule, and an antibody or antigen-binding fragment thereof.

248. The method of claim 246, wherein the integrin inhibitor is an antibody, an antigen-binding fragment, or a biosimilar thereof.

249. The method of claim 246, wherein the integrin inhibitor is a fusion protein or a biosimilar thereof.

250. The method of claim 246, wherein the integrin inhibitor is a peptidomimetic.

251. The method of claim 246, wherein the integrin inhibitor is selected from the group consisting of an 11 integrin inhibitor, an 21 integrin inhibitor, an IIb3 integrin inhibitor, an 41 (VLA-4) integrin inhibitor, an 47 integrin inhibitor, an 51 integrin inhibitor, an 53 integrin inhibitor, an 55 integrin inhibitor, an 56 integrin inhibitor, an E-selectin inhibitor, an ICAM-1 inhibitor, and a MAdCAM-1 inhibitor.

252. The method of claim 251, wherein the integrin inhibitor is an 47 integrin inhibitor.

253. The method of claim 252, wherein the 47 integrin inhibitor is selected from the group consisting of AJM300, vedolizumab, natalizumab and etrolizumab; and generic equivalents thereof.

254. The method of claim 252, wherein the 47 integrin inhibitor is vedolizumab, or a generic equivalent thereof.

255. The method of claim 230, wherein the ingestible device comprises an opening, and the method comprises delivering the immune modulator from the ingestible device via the opening.

256. The method of claim 255, wherein the opening comprises a nozzle.

257. The method of claim 256, wherein the ingestible device directly delivers the immune modulator to the GI tract of the subject via topical delivery.

258. The method of claim 256, wherein the ingestible device directly delivers the immune modulator to the GI tract of the subject via epithelial delivery.

259. The method of claim 256, wherein the ingestible device directly delivers the immune modulator to the GI tract of the subject via trans-epithelial delivery.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0109] FIG. 1 is a view of an example embodiment of an ingestible device, in accordance with some embodiments of the disclosure.

[0110] FIG. 2 is an exploded view of the ingestible device of FIG. 1, in accordance with some embodiments of the disclosure.

[0111] FIG. 3 is a diagram of an ingestible device during an example transit through a GI tract, in accordance with some embodiments of the disclosure.

[0112] FIG. 4 is a diagram of an ingestible device during an example transit through a jejunum, in accordance with some embodiments of the disclosure.

[0113] FIG. 5 is a flowchart of illustrative steps for determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0114] FIG. 6 is a flowchart of illustrative steps for detecting transitions from a stomach to a duodenum and from a duodenum back to a stomach, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0115] FIG. 7 is a plot illustrating data collected during an example operation of an ingestible device, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0116] FIG. 8 is another plot illustrating data collected during an example operation of an ingestible device, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0117] FIG. 9 is a flowchart of illustrative steps for detecting a transition from a duodenum to a jejunum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0118] FIG. 10 is a plot illustrating data collected during an example operation of an ingestible device, which may be used when detecting a transition from a duodenum to a jejunum, in accordance with some embodiments of the disclosure.

[0119] FIG. 11 is a plot illustrating muscle contractions detected by an ingestible device over time, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0120] FIG. 12 is a flowchart of illustrative steps for detecting a transition from a jejunum to an ileum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0121] FIG. 13 is a flowchart of illustrative steps for detecting a transition from a jejunum to an ileum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0122] FIG. 14 is a flowchart of illustrative steps for detecting a transition from an ileum to a cecum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0123] FIG. 15 is a flowchart of illustrative steps for detecting a transition from a cecum to a colon, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.

[0124] FIG. 16 illustrates an ingestible device for delivering a substance in the GI tract.

[0125] FIG. 17 illustrates aspects of a mechanism for an ingestible device with a gas generating cell configured to generate a gas to dispense a substance.

[0126] FIG. 18 illustrates an ingestible device having a piston to push for drug delivery.

[0127] FIG. 19 illustrates an ingestible device having a bellow structure for a storage reservoir of dispensable substances.

[0128] FIG. 20 illustrates an ingestible device having a flexible diaphragm to deform for drug delivery.

[0129] FIG. 21 shows an illustrative embodiment of an ingestible device with multiple openings in the housing.

[0130] FIG. 22 shows a highly cross-section of an ingestible device including a valve system and a sampling system.

[0131] FIG. 23 illustrates a valve system.

[0132] FIGS. 24A and 24B illustrate a portion of a two-stage valve system in its first and second stages, respectively.

[0133] FIGS. 25A and 25B illustrate a portion of a two-stage valve system in its first and second stages, respectively.

[0134] FIGS. 26A and 26B illustrate a portion of a two-stage valve system in its first and second stages, respectively.

[0135] FIG. 27 illustrates a more detailed view of an ingestible device including a valve system and a sampling system.

[0136] FIG. 28 illustrates a portion of an ingestible device including a sampling system and a two-stage valve system in its second stage.

[0137] FIG. 29 is a highly schematic illustrate of an ingestible device.

[0138] FIG. 30 is a graph shiwng the percentage (%) change in body weight at day 14 (SEM) for DSS mice treated with anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) daily (QD), when compared to mice treated with anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) every third day (Q3D) and vehicle control (Vehicle). Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0139] FIG. 31 is a graph showing the concentration of anti-IL-12 p40 rat IgG2A (g/mL) in plasma of anti-IL-12 p40 intraperitoneally (10 mg/kg) and intracecally (10 mg/kg and 1 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D) when compared to vehicle control (Vehicle) and when IP is compared to IC. ELISA analysis was used to determine the concentration of anti-IL-12 p40 (IgG2A). Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0140] FIG. 32 is a graph showing the concentration of anti-IL-12 p40 antibody (IgG2A) (g/mL) in the cecum and colon content of anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) and intracecally (10 mg/kg and 1 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. ELISA analysis was used to determine the concentration of rat IgG2A. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0141] FIG. 33 is a graph showing the mean overall tissue immunolabel scores (intensity and extent) in acute DSS colitis mouse colon of anti-IL-12 p40 antibody intracecally-treated versus vehicle control-treated DSS mice. Data presented as meanSEM.

[0142] FIG. 34 is a graph showing the mean location-specific immunolabel scores in acute DSS colitis mouse colon of anti-IL-12 p40 intracecally-treated versus vehicle control-treated DSS mice. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0143] FIG. 35 is a graph showing the ratio of anti-IL-12 p40 antibody in the colon tissue to the plasma concentration of the anti-IL-12 p40 antibody in mice treated with the anti-IL-12 p40 antibody on day 0 (Q0) or day 3 (Q3D) of the study, when measured at the same time point after the initial dosing. An outlier animal was removed from Group 5.

[0144] FIG. 36 is a graph showing the concentration of Il-1 (g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle). Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0145] FIG. 37 is a graph showing the concentration of 11-6 (g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle). Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.

[0146] FIG. 38 is a graph showing the concentration of Il-17A (g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg and 1 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle). Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0147] FIG. 39 is a graph showing the percentage (%) change in body weight at day 14 (SEM) for DSS mice treated with DATK32 (anti-47) antibody intraperitoneally (25 mg/kg) every third day (Q3D) or intracecally (25 mg/kg or 5 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle) and when IC is compared to IP. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0148] FIG. 40 is a graph showing the plasma concentration of DATK32 rat IgG2A (g/mL) of intraperitoneally (25 mg/kg) and intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0149] FIG. 41 is a graph showing the concentration of DATK32 rat IgG2A antibody (g/mL) in cecum and colon content of intraperitoneally (25 mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0150] FIG. 42 is a graph showing the concentration of DATK32 rat IgG2A (g/mL) in the colon content of intraperitoneally (25 mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD), and concentration over time (1, 2, 4, 24, and 48 hours), where IP is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0151] FIG. 43 is a graph showing the concentration of DATK32 rat IgG2A (g/g) in colon tissue of intraperitoneally (25 mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0152] FIG. 44 is a graph showing the concentration of DATK32 rat IgG2A (g/g) in the colon tissue of intraperitoneally (25 mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD), and the concentration over time (1, 2, 4, 24, and 48 hours) was determined, where IP is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0153] FIG. 45 is a graph showing the mean overall tissue immunolabel scores (intensity and extent) in acute DSS colitis mouse colon of DATK32 (anti-47) antibody treated versus vehicle control (Vehicle) treated DSS mice. The data are presented as meanSEM.

[0154] FIG. 46 is a graph showing the mean location-specific immunolabel scores in acute DSS colitis mouse colon of DATK32 (anti-47) antibody-treated versus vehicle control (Vehicle)-treated DSS mice. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0155] FIG. 47 is a graph showing the ratio of the DATK-32 antibody in the colon tissue to the plasma concentration of the DATK-32 antibody in mice treated with the DATK-32 antibody on day 0 (Q0) or day 3 (Q3D) of the study (Groups 9-12), when measured after initial dosing.

[0156] FIG. 48 is a graph showing the mean percentage of Th memory cells (meanSEM) in blood for DATK32 (anti-47) antibody intraperitoneally (25 mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. Mean percentage Th memory cells were measured using FACS analysis. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0157] FIG. 49 is an exemplary image of a histological section of a distal transverse colon of Animal 1501 showing no significant lesions (i.e., normal colon).

[0158] FIG. 50 is an exemplary image of a histological section of a distal transverse colon of Animal 2501 (treated with TNBS) showing areas of necrosis and inflammation.

[0159] FIG. 51 is a representative graph of plasma adalimumab concentrations over time following a single subcutaneous (SQ) or topical administration of adalimumab. The plasma concentrations of adalimumab were determined 6, 12, 24, and 48 hours after administration of adalimumab. N/D=not detectable.

[0160] FIG. 52 is a representative table of the plasma adalimumab concentrations (g/mL) as shown in FIG. 4.6.

[0161] FIG. 53 is a graph showing the concentration of TNF (pg/mL per mg of total protein) in non-inflamed and inflamed colon tissue after intracecal administration of adalimumab, as measured 6, 12, 24, and 24 hours after the initial dosing.

[0162] FIG. 54 is a graph showing the concentration of TNF (pg/mL per mg of total protein) in colon tissue after subcutaneous or intracecal (topical) administration of adalimumab, as measured 48 hours after the initial dosing.

[0163] FIG. 55 is a graph showing the percentage (%) change in body weight at day 14 (SEM) in acute DSS colitis mice treated with cyclosporine A orally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 3 mg/kg) daily (QD), when compared to vehicle control (Vehicle). Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0164] FIG. 56 is a graph showing the plasma cyclosporine A (CsA) (ng/mL) concentration over time (1 h, 2 h, 4 h, and 24 h) in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as meanSEM.

[0165] FIG. 57 is a graph showing the colon tissue cyclosporine A (CsA) (ng/g) concentration over time (1 h, 2 h, 4 h and 24 h) in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as meanSEM.

[0166] FIG. 58 is a graph showing the peak colon tissue cyclosporine A (CsA) (ng/g) concentration in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as meanSEM.

[0167] FIG. 59 is a graph showing the trough tissue concentration of cyclosporine (CsA) (ng/g) in colon of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as meanSEM.

[0168] FIG. 60 is a graph showing the interleukin-2 (Il-2) concentration (g/mL) in colon tissue of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA, where PO is compared to IC. Data presented as meanSEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0169] FIG. 61 is a graph showing the interleukin-6 (Il-6) concentration (g/mL) in colon tissue of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as meanSEM.

[0170] FIG. 62 illustrates a nonlimiting example of a system for collecting, communicating and/or analyzing data about a subject, using an ingestible device.

[0171] FIGS. 63A-63F are graphs showing rat IgG2A concentration as measured in (A) colon homogenate, (B) mLN homogenate, (C) small intestine homogenate, (D) cecum contents, (E) colon contents, and (F) plasma by ELISA. Standards were prepared with plasma matrix. Samples were diluted 1:50 before analysis. Sample 20 was removed from cecum contents analysis graph (outlier). *p<0.05; **p<0.01; ****p<0.001 were determined using the unpaired t test.

[0172] FIG. 64 illustrates a tapered silicon bellows.

[0173] FIG. 65 illustrates a tapered silicone bellows in the simulated device jig.

[0174] FIG. 66 illustrates a smooth PVC bellows.

[0175] FIG. 67 illustrates a smooth PVC bellows in the simulated device jig.

[0176] FIG. 68 demonstrates a principle of a competition assay performed in an experiment.

[0177] FIG. 69 shows AlphaLISA data.

[0178] FIG. 70 shows AlphaLISA data.

[0179] FIG. 71 shows AlphaLISA data.

[0180] FIG. 72 is a flowchart of illustrative steps of a clinical protocol, in accordance with some embodiments of the disclosure.

[0181] FIG. 73 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the cecum tissue of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.

[0182] FIG. 74 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the colon tissue of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.

[0183] FIG. 75 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the cecum contents of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.

[0184] FIG. 76 is a graph showing the mean concentration of tacrolimus in the cecum tissue and the proximal colon tissue 12 hours after intra-cecal or oral administration of tacrolimus to swine as described in Example 10.

[0185] FIG. 77 is a graph showing the mean concentration of tacrolimus in the blood 1 hour, 2 hours, 3 hours, 4 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.

[0186] FIG. 78 is a graph showing the AUC.sub.0-12 hours of tacrolimus in the blood after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13.

[0187] FIG. 79 is a graph showing the mean concentration of tacrolimus in the cecum tissue, the proximal colon tissue, the spiral colon tissue, the transverse colon tissue, and the distal colon tissue after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13. **** P<0.0001, *** P<0.001.

[0188] FIG. 80 is a graph showing the mean concentration of tacrolimus in the cecum lumen, the proximal lumen, the spiral colon lumen, the transverse colon lumen, and the distal colon lumen in swine after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13. **** P<0.0001, *** P<0.001

[0189] FIG. 81 is a bar graph showing the mean concentration of tacrolimus in the rectal content 1 hour, 3 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.

[0190] FIG. 82 is a line graph showing the mean concentration of tacrolimus in the rectal content 1 hour, 3 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.

[0191] FIG. 83 is a graph showing the mean concentration of a SMAD7 antisense molecule (SMAD7-AS-FAM) in the cecum tissue in untreated swine or in swine after intra-cecal (IC) or oral administration(PO) of SMAD7-AS-FAM as described in Example 9.

[0192] FIG. 84 is a graph showing the mean concentration of SMAD7-AS-FAM in the colon tissue in untreated swine or in swine after intra-cecal (IC) or oral administration(PO) of SMAD7-AS-FAM as described in Example 9.

[0193] FIG. 85 is a graph showing the mean concentration of SMAD7-AS-FAM in the colon contents in untreated swine or in swine after intra-cecal (IC) or oral administration(PO) of SMAD7-AS-FAM as described in Example 9.

[0194] FIG. 86 is a graph showing the mean concentration of SMAD7-AS-FAM in the cecum contents in untreated swine or in swine after intra-cecal (IC) or oral administration(PO) of SMAD7-AS-FAM as described in Example 9.

[0195] FIG. 87 is a graph showing the mean concentration of tacrolimus in the blood of swine 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

[0196] FIG. 88 is a graph showing the AUC.sub.0-12 hours of tacrolimus in the blood of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

[0197] FIG. 89 is a representative table showing the Tmax, Cmax, trough (at 12 hours post-administration), and AUC.sub.0-12 hours of tacrolimus in swine after intra-cecal (IC) or oral administration (PO) as described in Example 10.

[0198] FIG. 90 is a graph showing the mean concentration of tacrolimus in the cecum, the proximal colon, the spiral colon, the transverse colon, and the distal colon of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

[0199] FIG. 91 is a graph showing the mean concentration of tacrolimus in the cecum lumen, the proximal colon lumen, the spiral colon lumen, the transverse colon lumen, and the distal colon lumen of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

[0200] FIG. 92 is a graph showing the mean concentration of tacrolimus in the rectal content of swine at 1 hour, 3 hours, 6 hours, and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

[0201] FIG. 93 is a representative table showing the quantitative histological grading of colitis as described in Example 11.

[0202] FIG. 94 is a graph showing the histopathological scores of two slides for animal 1502 (healthy control swine treated with placebo), animal 2501 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), animal 2503 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), and animal 2504 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab) at the placebo or adalimumab administration site prior to administration of placebo or adalimumab, respectively. Absence of a bar for a particular parameter indicates that the value for this parameter was 0.

[0203] FIG. 95 is a representative hematoxylin- and eosin-stained image of the transverse colon of animal 1501 (healthy control swine). M, mucosa; SM, submucosa; TM, tunica muscularis. Numerous intestinal crypts (asterisks) are present and the surface epithelium (top two arrows) is intact. Mononuclear inflammatory cells are prominent in the lamina propria (light arrows) of the mucosa and extend a short distance into the submucosa (bottom two arrows). This amount of inflammatory cell infiltrate was expected background change and considered unrelated to the experimental protocol.

[0204] FIG. 96 is a representative hematoxylin- and eosin-stained image of the transverse colon of animal 2504 (8.5% DSS-induced colitis swine administered 1.86 mg/kg adalimumab) prior to administration of adalimumab. M, mucosa; SM, submucosa; TM, tunica muscularis. Extensive loss (light asterisks) of intestinal crypts is present in the mucosa. Scattered crypts remain (dark asterisks) and are often dilated and filled with inflammatory cell debris and mucus. The luminal epithelium persists in some areas (upper left arrow), but is absent in others (erosion; top middle and top right arrows). Inflammatory cells in the mucosa (light arrow) are abundant and extend into the submucosa (bottom left and bottom middle arrows).

[0205] FIG. 97 is a representative immunohistochemistry micrograph of the transverse colon of animal 1501 (healthy control swine) stained for human IgG. M, mucosa; SM, submucosa; TM, tunica muscularis. Serosal surface (arrows) and loose connective mesentery tissue (asterisks) are indicated. Faint 3,3-diaminobenzidine (DAB) staining in this tissue was considered a background effect and not indicative of human IgG.

[0206] FIG. 98 is a representative immunohistochemistry micrograph of the transverse colon of animal 2504 (8.5% DSS-induced colitis swine treated with 1.86 mg/kg dose of adsalimumab) stained for human IgG. M, mucosa; SM, submucosa; TM, tunica muscularis. DAB staining demonstrates the presence of human IgG at the surface of luminal epithelium (two top right arrows) and at the luminal surface of an area of inflammation and erosion (top two left arrows). Intense staining is also present in the loose connective mesentery tissue (asterisks) and extends a short distance into the outer edge of the tunica muscularis (bottom left two arrows). This type of staining was considered strong (grade 4) or very strong (grade 5).

[0207] FIG. 99 is a representative immunohistochemistry micrograph of the large intestine of animal 2504 (8.5% DSS-induced colitis swine treated with 1.86 mg/kg adalimumab) stained for human IgG. M, mucosa; SM, submucosa; TM, tunica muscularis. Lesions of DSS-induced colitis are present in this section. The luminal epithelium is absent (erosion) and diffuse loss of crypts (glands) is seen (top two asterisks). Very strong (grade 5) DAB (brown) staining demonstrates the presence of human IgG in the loose mesentery connective tissue (bottom two arterisks) and extending a short distance into the outer edge of the tunica muscularis (bottom two arrows). Strong (grade 4) staining for human IgG is seen at the eroded luminal surface (top two arrows pointing down) and within the inflammatory exudate. Weak (grade 2) staining for human IgG extends into the lamina propria (top two arrows pointing up) near the luminal surface.

[0208] FIG. 100 is a graph showing the presence of human IgG (adalimumab) at the specified locations (lumen/superficial mucosa, lamina propria, and tunica muscularis-outer/serosa) (scored level) in two slides from each of animal 1502 (placebo-treated healthy control swine), animal 2501 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), animal 2503 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab) and animal 2504 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab) at the placebo or adalimumab administration site. Absence of a bar for a particular location indicates that the value for this location was 0. Scoring: 0=not present; 1=minimal; 2=weak; 3=moderate; 4=strong; and 5=very strong immunolabel.

[0209] FIG. 101 is a graph showing the mean of Th memory cells (meanSEM) in Peyer's Patches (PP) for DATK32 antibody (anti-47 integrin antibody) intraperitoneally (25 mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. Mean Th memory cells were measured using FACS analysis. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0210] FIG. 102 is a graph showing the mean of Th memory cells (meanSEM) in mesenteric lymph nodes (mLN) for DATK32 antibody (anti-47 integrin antibody) intraperitoneally (25 mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. Mean Th memory cells were measured using FACS analysis. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p<0.05 was considered significant (Graph Pad Software, Inc.).

[0211] FIG. 103 is a graph showing the Disease Activity Index (DAI) of naive mice (Group 1), mice administered vehicle only both intraperitoneally (IP) and intracecally (IC) (Group 2), mice administered an anti-TNF antibody IP and vehicle IC (Group 7), and mice administered an anti-TNF antibody IC and vehicle IP (Group 8) at Day 28 and Day 42 of the study described in Example 16.

[0212] FIG. 104 is a set of graphs showing the colonic tissue concentration of TNF, IL-17A, IL-4, and IL-22 in mice administered vehicle only both IP and IC (Group 2), mice administered IgG control antibody IP and vehicle IC (Group 3), mice administered IgG control IC and vehicle IP (Group 4), mice administered anti-TNF antibody IP and vehicle IC (Group 7), and mice administered anti-TNF antibody IC and vehicle IP (Group 8) at Day 42 of the study described in Example 16.

[0213] FIG. 105 is a graph showing the Disease Activity Index (DAI) of naive mice (Group 1), mice administered vehicle only both IP and IC (Group 2), mice administered an anti-IL12 p40 antibody IP and vehicle IC (Group 5), and mice administered an anti-IL12 p40 antibody IC and vehicle IP (Group 6) at Day 28 and Day 42 of the study described in Example 16.

[0214] FIG. 106 is a set of graphs showing the colonic tissue concentration of IFNgamma, IL-6, IL-17A, TNF, IL-22, and IL-1b in nave mice (Group 1), mice administered vehicle only both IP and IC (Group 2), mice administered anti-IL12 p40 antibody IP and vehicle IC (Group 5), and mice administered anti-IL12 p40 antibody IC and vehicle IP (Group 8) at Day 42 of the study described in Example 16.

DETAILED DESCRIPTION

[0215] The present disclosure is directed to various methods and formulations for treating diseases of the gastrointestinal tract with a therapeutic agent as disclosed herein. For example, in an embodiment, a method of treating a disease of the gastrointestinal tract in a subject comprises administering to the subject a pharmaceutical formulation comprising a therapeutic agent as disclosed herein wherein the pharmaceutical formulation is released in the subject's gastrointestinal tract proximate to one or more sites of disease. For example, in an embodiment, the pharmaceutical formulation comprises a therapeutically effective amount of a therapeutic agent as disclosed herein.

[0216] In some embodiments, the formulation is contained in an ingestible device, and the device releases the formulation at a location proximate to the site of disease. The location of the site of disease may be predetermined. For example, an ingestible device, the location of which within the GI tract can be accurately determined as disclosed herein, may be used to sample one or more locations in the GI tract and to detect one or more analytes, including markers of the disease, in the GI tract of the subject. A pharmaceutical formulation may be then administered via an ingestible device and released at a location proximate to the predetermined site of disease. The release of the formulation may be triggered autonomously, as further described herein.

[0217] The following disclosure illustrates aspects of the formulations and methods embodied in the claims.

Formulations, Including Pharmaceutical Formulations

[0218] As used herein, a formulation of an immune modulator may refer to either the immune modulator in pure form such as, for example, the lyophilized immune modulator or a mixture of the immune modulator with one or more physiologically acceptable carriers, excipients or stabilizers. Thus, therapeutic formulations or medicaments can be prepared by mixing the immune modulator having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) antibody; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN, PLURONICS or polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers herein further include insterstitial drug dispersion agents such as soluble neutral-active hyaluronidase glycoproteins (sHASEGP), for example, human soluble PH-20 hyaluronidase glycoproteins, such as rHuPH20 (HYLENEX, Baxter International, Inc.). Certain exemplary sHASEGPs and methods of use, including rHuPH20, are described in US Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, a sHASEGP is combined with one or more additional glycosaminoglycanases such as chondroitinases. Exemplary lyophilized formulations are described in U.S. Pat. No. 6,267,958. Aqueous formulations include those described in U.S. Pat. No. 6,171,586 and WO2006/044908, the latter formulations including a histidine-acetate buffer.

[0219] A formulation of an immune modulator as disclosed herein, e.g., sustained-release formulations, can further include a mucoadhesive agent, e.g., one or more of polyvinyl pyrolidine, methyl cellulose, sodium carboxyl methyl cellulose, hydroxyl propyl cellulose, carbopol, a polyacrylate, chitosan, a eudragit analogue, a polymer, and a thiomer. Additional examples of mucoadhesive agents that can be included in a formulation with a therapeutic agent as disclosed herein are described in, e.g., Peppas et al., Biomaterials 17(16):1553-1561, 1996; Kharenko et al., Pharmaceutical Chemistry J. 43(4):200-208, 2009; Salamat-Miller et al., Adv. Drug Deliv. Reviews 57(11):1666-1691, 2005; Bernkop-Schnurch, Adv. Drug Deliv. Rev. 57(11):1569-1582, 2005; and Harding et al., Biotechnol. Genet. Eng. News 16(1):41-86, 1999.

[0220] In some embodiments, components of a formulation may include any one of the following components, or any combination thereof: Acacia, Alginate, Alginic Acid, Aluminum Acetate, an antiseptic, Benzyl Alcohol, Butyl Paraben, Butylated Hydroxy Toluene, an antioxidant. Citric acid, Calcium carbonate, Candelilla wax, a binder, Croscarmellose sodium, Confectioner sugar, Colloidal silicone dioxide, Cellulose, Carnuba wax, Corn starch, Carboxymethylcellulose calcium, Calcium stearate, Calcium disodium EDTA, Chelation agents, Copolyvidone, Castor oil hydrogenated, Calcium hydrogen phosphate dehydrate, Cetylpyridine chloride, Cysteine HCl, Crosspovidone, Dibasic Calcium Phosphate, Disodium hydrogen phosphate, Dimethicone, Erythrosine Sodium, Ethyl Cellulose, Gelatin, Glyceryl monooleate, Glycerin, Glycine, Glyceryl monostearate, Glyceryl behenate, Hydroxy propyl cellulose, Hydroxyl propyl methyl cellulose, Hypromellose, HPMC Pthalate, Iron oxides or ferric oxide, Iron oxide yellow, Iron oxide red or ferric oxide, Lactose (hydrous or anhydrous or monohydrate or spray dried), Magnesium stearate, Microcrystalline cellulose, Mannitol, Methyl cellulose Magnesium carbonate, Mineral oil, Methacrylic acid copolymer, Magnesium oxide, Methyl paraben, PEG, Polysorbate 80, Propylene glycol, Polyethylene oxide, Propylene paraben, Polaxamer 407 or 188 or plain, Potassium bicarbonate, Potassium sorbate, Potato starch, Phosphoric acid, Polyoxy140 stearate, Sodium starch glycolate, Starch pregelatinized, Sodium crossmellose, Sodium lauryl sulfate, Starch, Silicon dioxide, Sodium benzoate Stearic acid, Sucrose base for medicated confectionery, a granulating agent, Sorbic acid, Sodium carbonate, Saccharin sodium, Sodium alginate, Silica gel, Sorbiton monooleate, Sodium stearyl fumarate, Sodium chloride, Sodium metabisulfite, Sodium citrate dehydrate, Sodium starch, Sodium carboxy methyl cellulose, Succinic acid, Sodium propionate, Titanium dioxide, Talc, Triacetin, Triethyl citrate.

[0221] Accordingly, in some embodiments of the method of treating a disease as disclosed herein, the method comprises administering to the subject a pharmaceutical composition that is a formulation as disclosed herein. In some embodiments the formulation is a dosage form, which may be, as an example, a solid form such as, for example, a capsule, a tablet, a sachet, or a lozenge; or which may be, as an example, a liquid form such as, for example, a solution, a suspension, an emulsion, or a syrup.

[0222] In some embodiments the formulation is not comprised in an ingestible device. In some embodiments wherein the formulation is not comprised in an ingestible device, the formulation may be suitable for oral administration. The formulation may be, for example, a solid dosage form or a liquid dosage form as disclosed herein. In some embodiments wherein the formulation is not comprised in an ingestible device, the formulation may be suitable for rectal administration. The formulation may be, for example, a dosage form such as a suppository or an enema. In embodiments where the formulation is not comprised in an ingestible device, the formulation releases the immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release in the GI tract. Such localized release may be achieved, for example, with a formulation comprising an enteric coating. Such localized release may be achieved, an another example, with a formulation comprising a core comprising one or more polymers suitable for controlled release of an active substance. A non-limiting list of such polymers includes: poly(2-(diethylamino)ethyl methacrylate, 2-(dimethylamino)ethyl methacrylate, poly(ethylene glycol), poly(-aminoethyl methacrylate), (2-hydroxypropyl)methacrylamide, poly((3-benzyl-1-aspartate), poly(N-isopropylacrylamide), and cellulose derivatives.

[0223] In some embodiments the formulation is comprised in an ingestible device as disclosed herein. In some embodiments wherein the formulation is comprised in an ingestible device, the formulation may be suitable for oral administration. The formulation may be, for example, a solid dosage form or a liquid dosage form as disclosed herein. In some embodiments the formulation is suitable for introduction and optionally for storage in the device. In some embodiments the formulation is suitable for introduction and optionally for storage in the reservoir comprised in the device. In some embodiments the formulation is suitable for introduction and optionally for storage in the reservoir comprised in the device. Thus, in some embodiments, provided herein is a reservoir comprising a therapeutically effective amount of an immune modulator, wherein the reservoir is configured to fit into an ingestible device. In some embodiments, the reservoir comprising a therapeutically effective amount of an immune modulator is attachable to an ingestible device. In some embodiments, the reservoir comprising a therapeutically effective amount of an immune modulator is capable of anchoring itself to the subject's tissue. As an example, the reservoir capable of anchoring itself to the subject's tissue comprises silicone. As an example, the reservoir capable of anchoring itself to the subject's tissue comprises polyvinyl chloride.

[0224] In some embodiments the formulation is suitable for introduction in the spray catheters disclosed herein.

[0225] The formulation/medicament herein may also contain more than one active compound as necessary for the particular indication being treated, for example, those with complementary activities that do not adversely affect each other. For instance, the formulation may further comprise another immune modulator or a chemotherapeutic agent. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.

[0226] The active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

[0227] The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

[0228] Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the immune modulator, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-()-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated immune modulators remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37 C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular SS bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.

[0229] Pharmaceutical formulations may contain one or more immune modulators. The pharmaceutical formulations may be formulated in any manner known in the art. In some embodiments the formulations include one or more of the following components: a sterile diluent (e.g., sterile water or saline), a fixed oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents, antibacterial or antifungal agents, such as benzyl alcohol or methyl parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like, antioxidants, such as ascorbic acid or sodium bisulfite, chelating agents, such as ethylenediaminetetraacetic acid, buffers, such as acetates, citrates, or phosphates, and isotonic agents, such as sugars (e.g., dextrose), polyalcohols (e.g., mannitol or sorbitol), or salts (e.g., sodium chloride), or any combination thereof. Liposomal suspensions can also be used as pharmaceutically acceptable carriers (see, e.g., U.S. Pat. No. 4,522,811, incorporated by reference herein in its entirety). The formulations can be formulated and enclosed in ampules, disposable syringes, or multiple dose vials. Where required, proper fluidity can be maintained by, for example, the use of a coating, such as lecithin, or a surfactant. Controlled release of the immune modulator can be achieved by implants and microencapsulated delivery systems, which can include biodegradable, biocompatible polymers (e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid; Alza Corporation and Nova Pharmaceutical, Inc.).

[0230] In some embodiments, the immune modulator is present in a pharmaceutical formulation within the device.

[0231] In some embodiments, the immune modulator is present in solution within the device.

[0232] In some embodiments, the immune modulator is present in a suspension in a liquid medium within the device.

[0233] In some embodiments, the therapeutic agent as disclosed herein is present as a pure, powder (e.g., lyophilized) form of the therapeutic agent as disclosed herein.

Definitions

[0234] By ingestible, it is meant that the device can be swallowed whole.

[0235] The terms antibody and immunoglobulin are used interchangeably in the broadest sense and include monoclonal antibodies (for example, full length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific, trispecific etc. antibodies so long as they exhibit the desired biological activity) and may also include certain antibody fragments (as described in greater detail herein). An antibody can be human, humanized and/or affinity matured.

[0236] Antibody fragments comprise only a portion of an intact antibody, where in certain embodiments, the portion retains at least one, and typically most or all, of the functions normally associated with that portion when present in an intact antibody. In one embodiment, an antibody fragment comprises an antigen binding site of the intact antibody and thus retains the ability to bind antigen. In another embodiment, an antibody fragment, for example one that comprises the Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody, such as FcRn binding, antibody half-life modulation, ADCC function and complement binding. In one embodiment, an antibody fragment is a monovalent antibody that has an in vivo half-life substantially similar to an intact antibody. For example, such an antibody fragment may comprise on antigen binding arm linked to an Fc sequence capable of conferring in vivo stability to the fragment.

[0237] The term monoclonal antibody as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigen. Furthermore, in contrast to polyclonal antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.

[0238] The monoclonal antibodies herein specifically include chimeric antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).

[0239] Treatment regimen refers to a combination of dosage, frequency of administration, or duration of treatment, with or without addition of a second medication. Effective treatment regimen refers to a treatment regimen that will offer beneficial response to a patient receiving the treatment.

[0240] Effective amount refers to an amount of drug that offers beneficial response to a patient receiving the treatment. For example, an effective amount may be a Human Equivalent Dose (HED)

[0241] Dispensable, with reference to any substance, refers to any substance that may be released from an ingestible device as disclosed herein, or from a component of the device such as a reservoir. For example, a dispensable substance may be a therapeutic agent as disclosed herein, and/or a formulation comprising a therapeutic agent as disclosed herein.

[0242] Patient response or patient responsiveness can be assessed using any endpoint indicating a benefit to the patient, including, without limitation, (1) inhibition, to some extent, of disease progression, including slowing down and complete arrest; (2) reduction in the number of disease episodes and/or symptoms; (3) reduction in lesional size; (4) inhibition (i.e., reduction, slowing down or complete stopping) of disease cell infiltration into adjacent peripheral organs and/or tissues; (5) inhibition (i.e., reduction, slowing down or complete stopping) of disease spread; (6) decrease of auto-immune response, which may, but does not have to, result in the regression or ablation of the disease lesion; (7) relief, to some extent, of one or more symptoms associated with the disorder; (8) increase in the length of disease-free presentation following treatment; and/or (9) decreased mortality at a given point of time following treatment. The term responsiveness refers to a measurable response, including complete response (CR) and partial response (PR).

[0243] As used herein, complete response or CR means the disappearance of all signs of inflammation or remission in response to treatment. This does not necessarily mean the disease has been cured.

[0244] Partial response or PR refers to a decrease of at least 50% in the severity of inflammation, in response to treatment.

[0245] A beneficial response of a patient to treatment with a therapeutic agent and similar wording refers to the clinical or therapeutic benefit imparted to a patient at risk for or suffering from a inflammatory disease or condition that arises in a tissue originating from the endoderm. Such benefit includes cellular or biological responses, a complete response, a partial response, a stable disease (without progression or relapse), or a response with a later relapse of the patient from or as a result of the treatment with the agent.

[0246] As used herein, non-response or lack of response or similar wording means an absence of a complete response, a partial response, or a beneficial response to treatment with a therapeutic agent.

[0247] A patient maintains responsiveness to a treatment when the patient's responsiveness does not decrease with time during the course of a treatment.

[0248] A symptom of a disease or disorder (e.g., an inflammatory disease or condition that arises in tissue originating from the endoderm) is any morbid phenomenon or departure from the normal in structure, function, or sensation, experienced by a subject and indicative of disease.

[0249] Mucosa-associated lymphoid tissue or MALT refers to a diffuse system of small concentrations of lymphoid tissue found in various submucosal membrane sites of the body, such as the gastrointestinal tract, oral passage, nasopharyngeal tract, thyroid, breast, lung, salivary glands, eye, and skin.

[0250] Gut-associated lymphoid tissue or GALT refers to a part of the broader MALT and includes, e.g., Peyer's patches, mesenertic lymph nocdes, and isolated lymphoid follicles/intestinal lymphoid aggregates.

[0251] Peyer's patches refers to aggregated lymphoid modules organized into follicles and are important part of GALT. Peyer's patches are mainly present in the distal jejunum and the ileum.

[0252] Mesenteric lymph nodes refers to part of the paraaortic lymph node system that is a group of lymph nodes that lie between the layers of the mesentery and drain the gut tissues and deliver lymph to the thoracic duct. Mesenteric lymph nodes include the superior mesenteric lymph nodes which receive afferents from the jejunum, ileum, cecum, and the ascending and parts of the transverse colon. Mesenteric lymph nodes also include inferior mesenteric lymph nodes which are lymph nodes present throughout the hindgut. The hindgut, e.g., includes the distal third of the transverse colon and the splenic flexure, the descending colon, sigmoid colon, and the rectum. The lymph nodes drain into the superior mesenteric lymph nodes and ultimately to the preaortic lymph nodes.

[0253] Paraaortic lymph nodes refers to a group of mesenteric lymph nodes that lie in front of the lumbar vertebrae near the aorta. The paraaortic lymph nodes receive drainage from the gastrointestinal tract and the abdominal organs. Paraaortic lymph nodes include, e.g., retroaortic lymph nodes, lateral aortic lymph nodes, preaortic lymph nodes (e.g., Celiac, gastic, hepatic, and splenic lymph nodes), superior mesenteric lymph nodes (e.g., mesenteric, ileocolic, and mesocolic lymph nodes), and inferior mesenteric lymph nodes (e.g., pararectal lymph nodes).

[0254] As used herein, accuracy, when disclosed in connection with a specified location of a device within the GI tract of a subject, refers to the degree to which the location determined by the device conforms to the correct location, wherein the correct location is based on a generally accepted standard. The location within the GI tract of the subject determined by the device can be based on data, for example, light reflectance data, collected by the ingestible device. In some embodiments, the correct location can be based on external imaging devices, such as computer-aided tomography (CT), interpreted, for example, by a qualified clinician or physician. Therefore, percent accuracy (% accuracy) can refer to the percentage agreement between the location of the device in the GI tract as determined by the device, and the correct location, for example, as determined by CT, e.g., expressed as [(number of devices in which location determined by the device agrees with location as determined by CT/total devices administered to the subject or subjects)100%], or, where only one device is administered per subject, [(number of subjects in which location determined by the device agrees with location as determined by CT/total number of subjects)100%]. The latter formula for determining % accuracy was used in Example 14. In some embodiments, the accuracy with which the device determines a location refers to the accuracy with which the device determines that it is at a location pre-selected for drug release.

[0255] As used herein, an autonomous device refers to a device comprising one or more processors configured to independently control certain mechanisms or operations of the device while in the GI tract of a subject. Preferably, an autonomous device of the invention has no external electrical or wireless connections that control device mechanisms or operations, although connections such as wireless connections may be present to enable alternative device functions, such as transmitting data collected by the device to an external (ex vivo) system or receiver. The independently controlled mechanisms or operations of the autonomous device include, for example, triggering the release of a drug (or the formulation comprising the drug), triggering collection of one or more samples, and/or triggering the analysis of one or more samples; and/or determining the location of the device within the GI tract of the subject. Such a mechanism is referred to herein as an autonomous mechanism; for example, an autonomous triggering mechanism or an autonomous localization mechanism, respectively. Actively implementing such an autonomous triggering or localization mechanism is referred to as autonomous triggering or autonomous localizing, respectively. An autonomous localization mechanism is synonymous with a self-localization mechanism.

[0256] As used herein, a housing is a portion of an ingestible device that defines the boundary between the interior of the device and the environment exterior to the device.

[0257] As used herein, a self-localizing device refers to a device comprising a mechanism or system that can be implemented autonomously to determine the location of the ingestible device in vivo, e.g., within the GI tract of a subject. Such a mechanism is referred to as a self-localization mechanism. A self-localization mechanism is synonymous with an autonomous localization mechanism.A self-localizing device does not require ex vivo visualization devices or systems, for example, using scintigraphy or computer-aided tomography (CT), to localize in the GI tract.

[0258] As used herein, localizing the device refers to determining a location of the device.

[0259] As used herein, sensor refers to a mechanism or portion of a mechanism configured to collect information regarding the surroundings of the ingestible device. Examples of sensors include environmental sensors and light sensors. Examples of environmental sensors include pH sensors and sensors capable to identifying muscle contractions and/or peristalsis.

[0260] As used herein, time following transition refers to elapsed time after passage of the device from one portion, section or subsection of the GI tract into an adjacent portion, section or subsection of the GI tract.

[0261] As used herein, proximate as disclosed in connection with release of a drug from a device to one or more disease sites, refers to a location that is sufficiently spatially close to the one or more disease sites such that releasing the drug at the location treats the disease. For example, when the drug is released proximate to the one or more disease sites, the drug may be released 150 cm or less, such as 125 cm or less, such as 100 cm or less, such as 50 cm or less, such as 40 cm or less, such as 30 cm or less, such as 20 cm or less, such as 10 cm or less, such as 5 cm or less, such as 2 cm or less, from the one or more sites of disease. The proximate location for drug release may be in the same section or subsection of the gastrointestinal tract as the one or more disease sites. In the alternative, the proximate location for drug release may be in a different section or subsection of the GI tract than the one or more disease sites; for example, the drug release may be proximal to the one or more disease sites. In a non-limiting example, the drug may be released in the cecum to treat a site of disease tissue in the ascending colon (i.e., distal to the cecum). In another non-limiting example, the drug may be released in the cecum to treat a site of disease tissue in one or more of the ascending colon, transverse colon, descending colon, or rectum. Thus, where the present application refers to release of a drug proximate to a site of disease, this may in some embodiments refer to release in a section or subsection of the GI tract which has been determined to contain a site of disease. The section may be selected from esophagus, stomach, duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and rectum. The subsection may be selected from proximal duodenum, proximal jejunum, proximal ileum, proximal cecum, proximal ascending colon, proximal transverse colon, proximal descending colon, distal duodenum, distal jejunum, distal ileum, distal cecum, distal ascending colon, distal transverse colon, distal descending colon.

[0262] As used herein, the total induction dose is the sum of induction doses over a given time period.

[0263] As used herein, proximal, when used in connection with an anatomical structure, refers to a portion, section, or subsection that precedes, or is upstream of, an adjacent portion, section, or subsection of the anatomical structure. In some embodiments, proximal refers to a portion, section, or subsection that immediately precedes, or is immediately upstream of, an immediately adjacent portion, section, or subsection of the anatomical structure.

[0264] As used herein, distal, when used in connection with an anatomical structure, refers to a portion, section, or subsection that follows, or is downstream of, an adjacent portion, section, or subsection of the anatomical structure. In some embodiments, distal refers to a portion, section, or subsection that immediately follows, or is immediately downstream of, an immediately adjacent portion, section, or subsection of the anatomical structure.

[0265] As used herein, a reference to a drug's international nonproprietary name (INN) is to be interpreted as including generic, bioequivalent and biosimilar versions of that drug, including but not limited to any drug that has received abbreviated regulatory approval by reference to an earlier regulatory approval of that drug.

Inflammatory Conditions or Diseases that Arise from a Tissue Originating from the Endoderm

[0266] The presently claimed devices can, e.g., provide for a higher concentration of 47 expressing cells in the periphery (e.g., blood) when an immune modulator is delivered topically to one or more parts of the GI tract distal to the stomach (e.g., the small or large intestine) as compared to when the same dose of the immune modulator is systemically administered. The presently claimed devices can, e.g., result in trafficked cells being forced out of the local gastrointestinal tissue (including the mucosa) and lymph system, and back into systemic circulation of a subject.

[0267] Accordingly, also provided herein are methods of treating a disease or condition that arises in a tissue originating from the endoderm. The endoderm forms the gastrointestinal tract, respiratory tract, endocrine glands, and organs, the auditory system and urinary system. Thus, the present invention includes compositions and devices for treating diseases and conditions found in the following tissues that originate from the endoderm (e.g., the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder). Also provided herein are methods of treating a disease or a condition that arises in a tissue originating from the endoderm (e.g., any of the exemplary diseases or conditions that arise in a tissue originating from the endoderm described herein) that include intrathecally releasing one or more immune modulators in the small or large intestine using any of the devices or compositions described herein.

[0268] Non-limiting examples of a disease or condition that arises in a tissue originating from the endoderm includes gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjgren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radiation-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis. Additional examples of diseases and conditions that arise in a tissue originating from the endoderm are known in the are known in the art.

[0269] As used herein, the term immune modulator means a therapeutic agent that decreases the activation of an immune cell (e.g., a T cell, e.g., memory T cell), decreases the secretion or expression of a pro-inflammatory cytokine, decreases the recruitment or migration of T-lymphocytes (e.g., memory T lymphocytes), and/or increases the secretion or expression of an anti-inflammatory cytokine. Non-limiting examples of immune modulators are anti-inflammatory agents. Non-limiting examples of anti-inflammatory agents include IL-12/IL-23 inhibitors, TNF inhibitors, IL-6 receptor inhibitors, immune modulatory agents (e.g., CD40/CD40L inhibitors), IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, chemokine/chemokine receptor inhibitors, and integrin inhibitors. Non-limiting examples of integrin inhibitors include 137 integrin inhibitors, such as 47 integrin inhibitors. In some embodiments of any of the methods described herein, the immune modulator is a PDE4 inhibitor.

[0270] As used herein, the term immune modulator means a therapeutic agent that decreases the activation of an immune cell, decreases the secretion or expression of a pro-inflammatory cytokine, decreases the recruitment or migration of T-lymphocytes (e.g., memory T lymphocytes), and/or increases the secretion or expression of an anti-inflammatory cytokine. Non-limiting examples of immune modulators are anti-inflammatory agents. Non-limiting examples of anti-inflammatory agents include IL-12/IL-23 inhibitors, TNF inhibitors, IL-6 receptor inhibitors, immune modulatory agents (e.g., CD40/CD40L inhibitors), IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, chemokine/chemokine receptor inhibitors, and integrin inhibitors. In some embodiments of any of the methods described herein, the immune modulator is a PDE4 inhibitor. Additional examples of immune modulators useful for the treatment of a liver disease or disorder are described below.

[0271] Non-limiting exemplary examples of immune modulators are described below. Additional examples of immune modulators are known in the art.

IL-12/IL-23 Inhibitors

[0272] The term IL-12/IL-23 inhibitors refers to an agent which decreases IL-12 or IL-23 expression and/or the ability of IL-12 to bind to an IL-12 receptor or the ability of IL-23 to bind to an IL-23 receptor. IL-12 is a heterodimeric cytokine that includes both IL-12A (p35) and IL-12B (p40) polypeptides. IL-23 is a heterodimeric cytokine that includes both IL-23 (p19) and IL-12B (p40) polypeptides. The receptor for IL-12 is a heterodimeric receptor includes IL-12R (31 and IL-12R (32. The receptor for IL-23 receptor is a heterodimeric receptor that includes both IL-12R (31 and IL-23R.

[0273] In some embodiments, the IL-12/IL-23 inhibitor can decrease the binding of IL-12 to the receptor for IL-12. In some embodiments, the IL-12/IL-23 inhibitor can decrease the binding of IL-23 to the receptor for IL-23. In some embodiments, the IL-12/IL-23 inhibitor decreases the expression of IL-12 or IL-23. In some embodiments, the IL-12/IL-23 inhibitor decreases the expression of a receptor for IL-12. In some embodiments, the IL-12/IL-23 inhibitor decreases the expression of a receptor for IL-23.

[0274] In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-12B (p40) subunit. In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-12A (p35). In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-23 (p19). In some embodiments, the IL-12/IL-23 inhibitory agent targets the receptor for IL-12 (one or both of IL-12R 1 or IL-12R 2). In some embodiments, the IL-12/IL-23 inhibitory agent targets the receptor for IL-23 (one or both of IL-12R 1 and IL-23R).

[0275] In some embodiments, an IL-12/IL-23 inhibitor can be an inhibitory nucleic acid. In some embodiments, the inhibitory nucleic acid can be an antisense nucleic acid, a ribozyme, and a small interfering RNA (siRNA). Examples of aspects of these different oligonucleotides are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA in a mammalian cell can be synthesized in vitro.

[0276] Inhibitory nucleic acids that can decrease the expression of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 1-12).

TABLE-US-00001 HumanIL-12A(p35)mRNA (SEQIDNO:1) 1 tttcgctttcattttgggccgagctggaggcggcggggccgtcccggaacggctgcggcc 61 gggcaccccgggagttaatccgaaagcgccgcaagccccgcgggccggccgcaccgcacg 121 tgtcaccgagaagctgatgtagagagagacacagaaggagacagaaagcaagagaccaga 181 gtcccgggaaagtcctgccgcgcctcgggacaattataaaaatgtggccccctgggtcag 241 cctcccagccaccgccctcacctgccgcggccacaggtctgcatccagcggctcgccctg 301 tgtccctgcagtgccggctcagcatgtgtccagcgcgcagcctcctccttgtggctaccc 361 tggtcctcctggaccacctcagtttggccagaaacctccccgtggccactccagacccag 421 gaatgttcccatgccttcaccactcccaaaacctgctgagggccgtcagcaacatgctcc 481 agaaggccagacaaactctagaattttacccttgcacttctgaagagattgatcatgaag 541 atatcacaaaagataaaaccagcacagtggaggcctgtttaccattggaattaaccaaga 601 atgagagttgcctaaattccagagagacctctttcataactaatgggagttgcctggcct 661 ccagaaagacctcttttatgatggccctgtgccttagtagtatttatgaagacttgaaga 721 tgtaccaggtggagttcaagaccatgaatgcaaagcttctgatggatcctaagaggcaga 781 tctttctagatcaaaacatgctggcagttattgatgagctgatgcaggccctgaatttca 841 acagtgagactgtgccacaaaaatcctcccttgaagaaccggatttttataaaactaaaa 901 tcaagctctgcatacttcttcatgctttcagaattcgggcagtgactattgatagagtga 961 tgagctatctgaatgcttcctaaaaagcgaggtccctccaaaccgttgtcatttttataa 1021 aactttgaaatgaggaaactttgataggatgtggattaagaactagggagggggaaagaa 1081 ggatgggactattacatccacatgatacctctgatcaagtatttttgacatttactgtgg 1141 ataaattgtttttaagttttcatgaatgaattgctaagaagggaaaatatccatcctgaa 1201 ggtgtttttcattcactttaatagaagggcaaatatttataagctatttctgtaccaaag 1261 tgtttgtggaaacaaacatgtaagcataacttattttaaaatatttatttatataacttg 1321 gtaatcatgaaagcatctgagctaacttatatttatttatgttatatttattaaattatt 1381 tatcaagtgtatttgaaaaatatttttaagtgttctaaaaataaaagtattgaattaaag 1441 tgaaaaaaaa HumanIL-12B(p40)mRNA (SEQIDNO:2) 1 ctgtttcagggccattggactctccgtcctgcccagagcaagatgtgtcaccagcagttg 61 gtcatctcttggttttccctggtttttctggcatctcccctcgtggccatatgggaactg 121 aagaaagatgtttatgtcgtagaattggattggtatccggatgcccctggagaaatggtg 181 gtcctcacctgtgacacccctgaagaagatggtatcacctggaccttggaccagagcagt 241 gaggtcttaggctctggcaaaaccctgaccatccaagtcaaagagtttggagatgctggc 301 cagtacacctgtcacaaaggaggcgaggttctaagccattcgctcctgctgcttcacaaa 361 aaggaagatggaatttggtccactgatattttaaaggaccagaaagaacccaaaaataag 421 acctttctaagatgcgaggccaagaattattctggacgtttcacctgctggtggctgacg 481 acaatcagtactgatttgacattcagtgtcaaaagcagcagaggctcttctgacccccaa 541 ggggtgacgtgcggagctgctacactctctgcagagagagtcagaggggacaacaaggag 601 tatgagtactcagtggagtgccaggaggacagtgcctgcccagctgctgaggagagtctg 661 cccattgaggtcatggtggatgccgttcacaagctcaagtatgaaaactacaccagcagc 721 ttcttcatcagggacatcatcaaacctgacccacccaagaacttgcagctgaagccatta 781 aagaattctcggcaggtggaggtcagctgggagtaccctgacacctggagtactccacat 841 tcctacttctccctgacattctgcgttcaggtccagggcaagagcaagagagaaaagaaa 901 gatagagtcttcacggacaagacctcagccacggtcatctgccgcaaaaatgccagcatt 961 agcgtgcgggcccaggaccgctactatagctcatcttggagcgaatgggcatctgtgccc 1021 tgcagttaggttctgatccaggatgaaaatttggaggaaaagtggaagatattaagcaaa 1081 atgtttaaagacacaacggaatagacccaaaaagataatttctatctgatttgctttaaa 1141 acgtttttttaggatcacaatgatatctttgctgtatttgtatagttagatgctaaatgc 1201 tcattgaaacaatcagctaatttatgtatagattttccagctctcaagttgccatgggcc 1261 ttcatgctatttaaatatttaagtaatttatgtatttattagtatattactgttatttaa 1321 cgtttgtctgccaggatgtatggaatgtttcatactcttatgacctgatccatcaggatc 1381 agtccctattatgcaaaatgtgaatttaattttatttgtactgacaacttttcaagcaag 1441 gctgcaagtacatcagttttatgacaatcaggaagaatgcagtgttctgataccagtgcc 1501 atcatacacttgtgatggatgggaacgcaagagatacttacatggaaacctgacaatgca 1561 aacctgttgagaagatccaggagaacaagatgctagttcccatgtctgtgaagacttcct 1621 ggagatggtgttgataaagcaatttagggccacttacacttctaagcaagtttaatcttt 1681 ggatgcctgaattttaaaagggctagaaaaaaatgattgaccagcctgggaaacataaca 1741 agaccccgtctctacaaaaaaaatttaaaattagccaggcgtggtggctcatgcttgtgg 1801 tcccagctgttcaggaggatgaggcaggaggatctcttgagcccaggaggtcaaggctat 1861 ggtgagccgtgattgtgccactgcataccagcctaggtgacagaatgagaccctgtctca 1921 aaaaaaaaaatgattgaaattaaaattcagctttagcttccatggcagtcctcaccccca 1981 cctctctaaaagacacaggaggatgacacagaaacaccgtaagtgtctggaaggcaaaaa 2041 gatcttaagattcaagagagaggacaagtagttatggctaaggacatgaaattgtcagaa 2101 tggcaggtggcttcttaacagccctgtgagaagcagacagatgcaaagaaaatctggaat 2161 ccctttctcattagcatgaatgaacctgatacacaattatgaccagaaaatatggctcca 2221 tgaaggtgctacttttaagtaatgtatgtgcgctctgtaaagtgattacatttgtttcct 2281 gtttgtttatttatttatttatttttgcattctgaggctgaactaataaaaactcttctt 2341 tgtaatc HumanIL-23(p19)mRNA (SEQIDNO:3) 1 aaaacaacaggaagcagcttacaaactcggtgaacaactgagggaaccaaaccagagacg 61 cgctgaacagagagaatcaggctcaaagcaagtggaagtgggcagagattccaccaggac 121 tggtgcaaggcgcagagccagccagatttgagaagaaggcaaaaagatgctggggagcag 181 agctgtaatgctgctgttgctgctgccctggacagctcagggcagagctgtgcctggggg 241 cagcagccctgcctggactcagtgccagcagctttcacagaagctctgcacactggcctg 301 gagtgcacatccactagtgggacacatggatctaagagaagagggagatgaagagactac 361 aaatgatgttccccatatccagtgtggagatggctgtgacccccaaggactcagggacaa 421 cagtcagttctgcttgcaaaggatccaccagggtctgattttttatgagaagctgctagg 481 atcggatattttcacaggggagccttctctgctccctgatagccctgtgggccagcttca 541 tgcctccctactgggcctcagccaactcctgcagcctgagggtcaccactgggagactca 601 gcagattccaagcctcagtcccagccagccatggcagcgtctccttctccgcttcaaaat 661 ccttcgcagcctccaggcctttgtggctgtagccgcccgggtctttgcccatggagcagc 721 aaccctgagtccctaaaggcagcagctcaaggatggcactcagatctccatggcccagca 781 aggccaagataaatctaccaccccaggcacctgtgagccaacaggttaattagtccatta 841 attttagtgggacctgcatatgttgaaaattaccaatactgactgacatgtgatgctgac 901 ctatgataaggttgagtatttattagatgggaagggaaatttggggattatttatcctcc 961 tggggacagtttggggaggattatttattgtatttatattgaattatgtacttttttcaa 1021 taaagtcttatttttgtggctaaaaaaaa HumanIL-12R1mRNAVariant1 (SEQIDNO:4) 1 ctctttcactttgacttgccttagggatgggctgtgacactttactttttttcttttttc 61 ttttttttcagtcttttctccttgctcagcttcaatgtgttccggagtggggacggggtg 121 gctgaacctcgcaggtggcagagaggctcccctggggctgtggggctctacgtggatccg 181 atggagccgctggtgacctgggtggtccccctcctcttcctcttcctgctgtccaggcag 241 ggcgctgcctgcagaaccagtgagtgctgttttcaggacccgccatatccggatgcagac 301 tcaggctcggcctcgggccctagggacctgagatgctatcggatatccagtgatcgttac 361 gagtgctcctggcagtatgagggtcccacagctggggtcagccacttcctgcggtgttgc 421 cttagctccgggcgctgctgctacttcgccgccggctcagccaccaggctgcagttctcc 481 gaccaggctggggtgtctgtgctgtacactgtcacactctgggtggaatcctgggccagg 541 aaccagacagagaagtctcctgaggtgaccctgcagctctacaactcagttaaatatgag 601 cctcctctgggagacatcaaggtgtccaagttggccgggcagctgcgtatggagtgggag 661 accccggataaccaggttggtgctgaggtgcagttccggcaccggacacccagcagccca 721 tggaagttgggcgactgcggacctcaggatgatgatactgagtcctgcctctgccccctg 781 gagatgaatgtggcccaggaattccagctccgacgacggcagctggggagccaaggaagt 841 tcctggagcaagtggagcagccccgtgtgcgttccccctgaaaaccccccacagcctcag 901 gtgagattctcggtggagcagctgggccaggatgggaggaggcggctgaccctgaaagag 961 cagccaacccagctggagcttccagaaggctgtcaagggctggcgcctggcacggaggtc 1021 acttaccgactacagctccacatgctgtcctgcccgtgtaaggccaaggccaccaggacc 1081 ctgcacctggggaagatgccctatctctcgggtgctgcctacaacgtggctgtcatctcc 1141 tcgaaccaatttggtcctggcctgaaccagacgtggcacattcctgccgacacccacaca 1201 gaaccagtggctctgaatatcagcgtcggaaccaacgggaccaccatgtattggccagcc 1261 cgggctcagagcatgacgtattgcattgaatggcagcctgtgggccaggacgggggcctt 1321 gccacctgcagcctgactgcgccgcaagacccggatccggctggaatggcaacctacagc 1381 tggagtcgagagtctggggcaatggggcaggaaaagtgttactacattaccatctttgcc 1441 tctgcgcaccccgagaagctcaccttgtggtctacggtcctgtccacctaccactttggg 1501 ggcaatgcctcagcagctgggacaccgcaccacgtctcggtgaagaatcatagcttggac 1561 tctgtgtctgtggactgggcaccatccctgctgagcacctgtcccggcgtcctaaaggag 1621 tatgttgtccgctgccgagatgaagacagcaaacaggtgtcagagcatcccgtgcagccc 1681 acagagacccaagttaccctcagtggcctgcgggctggtgtagcctacacggtgcaggtg 1741 cgagcagacacagcgtggctgaggggtgtctggagccagccccagcgcttcagcatcgaa 1801 gtgcaggtttctgattggctcatcttcttcgcctccctggggagcttcctgagcatcctt 1861 ctcgtgggcgtccttggctaccttggcctgaacagggccgcacggcacctgtgcccgccg 1921 ctgcccacaccctgtgccagctccgccattgagttccctggagggaaggagacttggcag 1981 tggatcaacccagtggacttccaggaagaggcatccctgcaggaggccctggtggtagag 2041 atgtcctgggacaaaggcgagaggactgagcctctcgagaagacagagctacctgagggt 2101 gcccctgagctggccctggatacagagttgtccttggaggatggagacaggtgcaaggcc 2161 aagatgtgatcgttgaggctcagagagggtgagtgactcgcccgaggctacgtagcacac 2221 acaggagtcacatttggacccaaataacccagagctcctccaggctccagtgcacctgcc 2281 tcctctctgccccgtgcctgttgccacccatcctgcgggggaaccctagatgctgccatg 2341 aaatggaagctgctgcaccctgctgggcctggcatccgtggggcaggagcagaccctgcc 2401 atttacctgttctggcgtagaatggactgggaatgggggcaaggggggctcagatggatc 2461 cctggaccctgggctgggcatccacccccaggagcactggatggggagtctggactcaag 2521 ggctccctgcagcattgcggggtcttgtagcttggaggatccaggcatatagggaagggg 2581 gctgtaaactttgtgggaaaaatgacggtcctcccatcccaccccccaccccaccctcac 2641 ccccctataaaatgggggtggtgataatgaccttacacagctgttcaaaatcatcgtaaa 2701 tgagcctcctcttgggtatttttttcctgtttgaagcttgaatgtcctgctcaaaatctc 2761 aaaacacgagccttggaattcaaaaaaaaaaaaaaaaaaa HumanIL-12R1mRNAVariant2 (SEQIDNO:5) 1 ctctttcactttgacttgccttagggatgggctgtgacactttactttttttcttttttc 61 ttttttttcagtcttttctccttgctcagcttcaatgtgttccggagtggggacggggtg 121 gctgaacctcgcaggtggcagagaggctcccctggggctgtggggctctacgtggatccg 181 atggagccgctggtgacctgggtggtccccctcctcttcctcttcctgctgtccaggcag 241 ggcgctgcctgcagaaccagtgagtgctgttttcaggacccgccatatccggatgcagac 301 tcaggctcggcctcgggccctagggacctgagatgctatcggatatccagtgatcgttac 361 gagtgctcctggcagtatgagggtcccacagctggggtcagccacttcctgcggtgttgc 421 cttagctccgggcgctgctgctacttcgccgccggctcagccaccaggctgcagttctcc 481 gaccaggctggggtgtctgtgctgtacactgtcacactctgggtggaatcctgggccagg 541 aaccagacagagaagtctcctgaggtgaccctgcagctctacaactcagttaaatatgag 601 cctcctctgggagacatcaaggtgtccaagttggccgggcagctgcgtatggagtgggag 661 accccggataaccaggttggtgctgaggtgcagttccggcaccggacacccagcagccca 721 tggaagttgggcgactgcggacctcaggatgatgatactgagtcctgcctctgccccctg 781 gagatgaatgtggcccaggaattccagctccgacgacggcagctggggagccaaggaagt 841 tcctggagcaagtggagcagccccgtgtgcgttccccctgaaaaccccccacagcctcag 901 gtgagattctcggtggagcagctgggccaggatgggaggaggcggctgaccctgaaagag 961 cagccaacccagctggagcttccagaaggctgtcaagggctggcgcctggcacggaggtc 1021 acttaccgactacagctccacatgctgtcctgcccgtgtaaggccaaggccaccaggacc 1081 ctgcacctggggaagatgccctatctctcgggtgctgcctacaacgtggctgtcatctcc 1141 tcgaaccaatttggtcctggcctgaaccagacgtggcacattcctgccgacacccacaca 1201 gatggcatgatctcagctcactgcaacctccgccttccagattcaagagattctcctgct 1261 tcagcctcccgagtagctgggattacaggcatctgccaccatacccggctaattttgtat 1321 ttttagtagagacggggtttcaccacgttggccaggctggtctcgaactcctgacctcaa 1381 gtgatccacctgccttggcctcccaaagtgttgggattataggcgtgagccaccatgccc 1441 agcctaatttttgtatttttagtagagatggagtttcaccatgttgcccaggctggtctc 1501 aaactcctgccctcaggtgatccacccacctcagcctctcaaagtgctgggattacaggt 1561 gtgagccactgtggccgacctactatttttattatttttgagctaggttctcagtctgtt 1621 ggcagactggagtgcaatcatggctcactgcagccttgaactcccagactcaagtgatcc 1681 ttccacctcagcctctggagtagctgggactacagacatgcaccaccacacctggttaat 1741 tttttatttttattttttgtagagacaggtgtctctctacgttgcccaggctggtctcga 1801 actcctgggctcaagtgatccacccatctccacctcccaaagtgctaggattacaggcgt 1861 gagccaccgtacccagcctggtcccatatcatagtgaaatggtgcctgtaaagctctcag 1921 cattggcttggcacatgcagttggtactcaataaacggctgttgctatccccaaaaaaaa 1981 aaaaaaaaaaaaaaaaa HumanIL-12R1mRNAVariant3 (SEQIDNO:6) 1 ctctttcactttgacttgccttagggatgggctgtgacactttactttttttcttttttc 61 ttttttttcagtcttttctccttgctcagcttcaatgtgttccggagtggggacggggtg 121 gctgaacctcgcaggtggcagagaggctcccctggggctgtggggctctacgtggatccg 181 atggagccgctggtgacctgggtggtccccctcctcttcctcttcctgctgtccaggcag 241 ggcgctgcctgcagaaccagtgagtgctgttttcaggacccgccatatccggatgcagac 301 tcaggctcggcctcgggccctagggacctgagatgctatcggatatccagtgatcgttac 361 gagtgctcctggcagtatgagggtcccacagctggggtcagccacttcctgcggtgttgc 421 cttagctccgggcgctgctgctacttcgccgccggctcagccaccaggctgcagttctcc 481 gaccaggctggggtgtctgtgctgtacactgtcacactctgggtggaatcctgggccagg 541 aaccagacagagaagtctcctgaggtgaccctgcagctctacaactcagttaaatatgag 601 cctcctctgggagacatcaaggtgtccaagttggccgggcagctgcgtatggagtgggag 661 accccggataaccaggttggtgctgaggtgcagttccggcaccggacacccagcagccca 721 tggaagttgggcgactgcggacctcaggatgatgatactgagtcctgcctctgccccctg 781 gagatgaatgtggcccaggaattccagctccgacgacggcagctggggagccaaggaagt 841 tcctggagcaagtggagcagccccgtgtgcgttccccctgaaaaccccccacagcctcag 901 gtgagattctcggtggagcagctgggccaggatgggaggaggcggctgaccctgaaagag 961 cagccaacccagctggagcttccagaaggctgtcaagggctggcgcctggcacggaggtc 1021 acttaccgactacagctccacatgctgtcctgcccgtgtaaggccaaggccaccaggacc 1081 ctgcacctggggaagatgccctatctctcgggtgctgcctacaacgtggctgtcatctcc 1141 tcgaaccaatttggtcctggcctgaaccagacgtggcacattcctgccgacacccacaca 1201 gaaccagtggctctgaatatcagcgtcggaaccaacgggaccaccatgtattggccagcc 1261 cgggctcagagcatgacgtattgcattgaatggcagcctgtgggccaggacgggggcctt 1321 gccacctgcagcctgactgcgccgcaagacccggatccggctggaatggcaacctacagc 1381 tggagtcgagagtctggggcaatggggcaggaaaagtgttactacattaccatctttgcc 1441 tctgcgcaccccgagaagctcaccttgtggtctacggtcctgtccacctaccactttggg 1501 ggcaatgcctcagcagctgggacaccgcaccacgtctcggtgaagaatcatagcttggac 1561 tctgtgtctgtggactgggcaccatccctgctgagcacctgtcccggcgtcctaaaggag 1621 tatgttgtccgctgccgagatgaagacagcaaacaggtgtcagagcatcccgtgcagccc 1681 acagagacccaagttaccctcagtggcctgcgggctggtgtagcctacacggtgcaggtg 1741 cgagcagacacagcgtggctgaggggtgtctggagccagccccagcgcttcagcatcgaa 1801 gtgcaggtttctgattggctcatcttcttcgcctccctggggagcttcctgagcatcctt 1861 ctcgtgggcgtccttggctaccttggcctgaacagggccgcacggcacctgtgcccgccg 1921 ctgcccacaccctgtgccagctccgccattgagttccctggagggaaggagacttggcag 1981 tggatcaacccagtggacttccaggaagaggcatccctgcaggaggccctggtggtagag 2041 atgtcctgggacaaaggcgagaggactgagcctctcgagaagacagagctacctgagggt 2101 gcccctgagctggccctggatacagagttgtccttggaggatggagacagatgtgatcgt 2161 tgaggctcagagagggtgagtgactcgcccgaggctacgtagcacacacaggagtcacat 2221 ttggacccaaataacccagagctcctccaggctccagtgcacctgcctcctctctgcccc 2281 gtgcctgttgccacccatcctgcgggggaaccctagatgctgccatgaaatggaagctgc 2341 tgcaccctgctgggcctggcatccgtggggcaggagcagaccctgccatttacctgttct 2401 ggcgtagaatggactgggaatgggggcaaggggggctcagatggatccctggaccctggg 2461 ctgggcatccacccccaggagcactggatggggagtctggactcaagggctccctgcagc 2521 attgcggggtcttgtagcttggaggatccaggcatatagggaagggggctgtaaactttg 2581 tgggaaaaatgacggtcctcccatcccaccccccaccccaccctcacccccctataaaat 2641 gggggtggtgataatgaccttacacagctgttcaaaatcatcgtaaatgagcctcctctt 2701 gggtatttttttcctgtttgaagcttgaatgtcctgctcaaaatctcaaaacacgagcct 2761 tggaattcaaaaaaaaaaaaaaaaaaa HumanIL-12R1mRNAVariant4 (SEQIDNO:7) 1 agaacactccgctgcctctccagagccaggcacacagcaggcgctccataaatgttcgtt 61 ggtcttttctccttgctcagcttcaatgtgttccggagtggggacggggtggctgaacct 121 cgcaggtggcagagaggctcccctggggctgtggggctctacgtggatccgatggagccg 181 ctggtgacctgggtggtccccctcctcttcctcttcctgctgtccaggcagggcgctgcc 241 tgcagaaccagtgagtgctgttttcaggacccgccatatccggatgcagactcaggctcg 301 gcctcgggccctagggacctgagatgctatcggatatccagtgatcgttacgagtgctcc 361 tggcagtatgagggtcccacagctggggtcagccacttcctgcggtgttgccttagctcc 421 gggcgctgctgctacttcgccgccggctcagccaccaggctgcagttctccgaccaggct 481 ggggtgtctgtgctgtacactgtcacactctgggtggaatcctgggccaggaaccagaca 541 gagaagtctcctgaggtgaccctgcagctctacaactcagttaaatatgagcctcctctg 601 ggagacatcaaggtgtccaagttggccgggcagctgcgtatggagtgggagaccccggat 661 aaccaggttggtgctgaggtgcagttccggcaccggacacccagcagcccatggaagttg 721 ggcgactgcggacctcaggatgatgatactgagtcctgcctctgccccctggagatgaat 781 gtggcccaggaattccagctccgacgacggcagctggggagccaaggaagttcctggagc 841 aagtggagcagccccgtgtgcgttccccctgaaaaccccccacagcctcaggtgagattc 901 tcggtggagcagctgggccaggatgggaggaggcggctgaccctgaaagagcagccaacc 961 cagctggagcttccagaaggctgtcaagggctggcgcctggcacggaggtcacttaccga 1021 ctacagctccacatgctgtcctgcccgtgtaaggccaaggccaccaggaccctgcacctg 1081 gggaagatgccctatctctcgggtgctgcctacaacgtggctgtcatctcctcgaaccaa 1141 tttggtcctggcctgaaccagacgtggcacattcctgccgacacccacacagaaccagtg 1201 gctctgaatatcagcgtcggaaccaacgggaccaccatgtattggccagcccgggctcag 1261 agcatgacgtattgcattgaatggcagcctgtgggccaggacgggggccttgccacctgc 1321 agcctgactgcgccgcaagacccggatccggctggaatggcaacctacagctggagtcga 1381 gagtctggggcaatggggcaggaaaagtgttactacattaccatctttgcctctgcgcac 1441 cccgagaagctcaccttgtggtctacggtcctgtccacctaccactttgggggcaatgcc 1501 tcagcagctgggacaccgcaccacgtctcggtgaagaatcatagcttggactctgtgtct 1561 gtggactgggcaccatccctgctgagcacctgtcccggcgtcctaaaggagtatgttgtc 1621 cgctgccgagatgaagacagcaaacaggtgtcagagcatcccgtgcagcccacagagacc 1681 caagttaccctcagtggcctgcgggctggtgtagcctacacggtgcaggtgcgagcagac 1741 acagcgtggctgaggggtgtctggagccagccccagcgcttcagcatcgaagtgcaggtt 1801 tctgattggctcatcttcttcgcctccctggggagcttcctgagcatccttctcgtgggc 1861 gtccttggctaccttggcctgaacagggccgcacggcacctgtgcccgccgctgcccaca 1921 ccctgtgccagctccgccattgagttccctggagggaaggagacttggcagtggatcaac 1981 ccagtggacttccaggaagaggcatccctgcaggaggccctggtggtagagatgtcctgg 2041 gacaaaggcgagaggactgagcctctcgagaagacagagctacctgagggtgcccctgag 2101 ctggccctggatacagagttgtccttggaggatggagacaggtgcaaggccaagatgtga 2161 tcgttgaggctcagagagggtgagtgactcgcccgaggctacgtagcacacacaggagtc 2221 acatttggacccaaataacccagagctcctccaggctccagtgcacctgcctcctctctg 2281 ccccgtgcctgttgccacccatcctgcgggggaaccctagatgctgccatgaaatggaag 2341 ctgctgcaccctgctgggcctggcatccgtggggcaggagcagaccctgccatttacctg 2401 ttctggcgtagaatggactgggaatgggggcaaggggggctcagatggatccctggaccc 2461 tgggctgggcatccacccccaggagcactggatggggagtctggactcaagggctccctg 2521 cagcattgcggggtcttgtagcttggaggatccaggcatatagggaagggggctgtaaac 2581 tttgtgggaaaaatgacggtcctcccatcccaccccccaccccaccctcacccccctata 2641 aaatgggggtggtgataatgaccttacacagctgttcaaaatcatcgtaaatgagcctcc 2701 tcttgggtatttttttcctgtttgaagcttgaatgtcctgctcaaaatctcaaaacacga 2761 gccttggaattcaaaaaaaaaaaaaaaaaaa HumanIL-12R2mRNAVariant1 (SEQIDNO:8) 1 tgcagagcacagagaaaggacatctgcgaggaaagttccctgatggctgtcaacaaagtg 61 ccacgtctctatggctgtgaacgctgagcacacgattttatcgcgcctatcatatcttgg 121 tgcataaacgcacctcacctcggtcaacccttgctccgtcttatgagacaggctttatta 181 tccgcattttatatgaggggaaactgacggtggagagagaattatcttgctcaaggcgac 241 acagcagagcccacaggtggcagaatcccacccgagcccgcttcgacccgcggggtggaa 301 accacgggcgcccgcccggctgcgcttccagagctgaactgagaagcgagtcctctccgc 361 cctgcggccaccgcccagccccgacccccgccccggcccgatcctcactcgccgccagct 421 ccccgcgcccaccccggagttggtggcgcagaggcgggaggcggaggcgggagggcgggc 481 gctggcaccgggaacgcccgagcgccggcagagagcgcggagagcgcgacacgtgcggcc 541 cagagcaccggggccacccggtccccgcaggcccgggaccgcgcccgctggcaggcgaca 601 cgtggaagaatacggagttctataccagagttgattgttgatggcacatacttttagagg 661 atgctcattggcatttatgtttataatcacgtggctgttgattaaagcaaaaatagatgc 721 gtgcaagagaggcgatgtgactgtgaagccttcccatgtaattttacttggatccactgt 781 caatattacatgctctttgaagcccagacaaggctgctttcactattccagacgtaacaa 841 gttaatcctgtacaagtttgacagaagaatcaattttcaccatggccactccctcaattc 901 tcaagtcacaggtcttccccttggtacaaccttgtttgtctgcaaactggcctgtatcaa 961 tagtgatgaaattcaaatatgtggagcagagatcttcgttggtgttgctccagaacagcc 1021 tcaaaatttatcctgcatacagaagggagaacaggggactgtggcctgcacctgggaaag 1081 aggacgagacacccacttatacactgagtatactctacagctaagtggaccaaaaaattt 1141 aacctggcagaagcaatgtaaagacatttattgtgactatttggactttggaatcaacct 1201 cacccctgaatcacctgaatccaatttcacagccaaggttactgctgtcaatagtcttgg 1261 aagctcctcttcacttccatccacattcacattcttggacatagtgaggcctcttcctcc 1321 gtgggacattagaatcaaatttcaaaaggcttctgtgagcagatgtaccctttattggag 1381 agatgagggactggtactgcttaatcgactcagatatcggcccagtaacagcaggctctg 1441 gaatatggttaatgttacaaaggccaaaggaagacatgatttgctggatctgaaaccatt 1501 tacagaatatgaatttcagatttcctctaagctacatctttataagggaagttggagtga 1561 ttggagtgaatcattgagagcacaaacaccagaagaagagcctactgggatgttagatgt 1621 ctggtacatgaaacggcacattgactacagtagacaacagatttctcttttctggaagaa 1681 tctgagtgtctcagaggcaagaggaaaaattctccactatcaggtgaccttgcaggagct 1741 gacaggagggaaagccatgacacagaacatcacaggacacacctcctggaccacagtcat 1801 tcctagaaccggaaattgggctgtggctgtgtctgcagcaaattcaaaaggcagttctct 1861 gcccactcgtattaacataatgaacctgtgtgaggcagggttgctggctcctcgccaggt 1921 ctctgcaaactcagagggcatggacaacattctggtgacttggcagcctcccaggaaaga 1981 tccctctgctgttcaggagtacgtggtggaatggagagagctccatccagggggtgacac 2041 acaggtccctctaaactggctacggagtcgaccctacaatgtgtctgctctgatttcaga 2101 gaacataaaatcctacatctgttatgaaatccgtgtgtatgcactctcaggggatcaagg 2161 aggatgcagctccatcctgggtaactctaagcacaaagcaccactgagtggcccccacat 2221 taatgccatcacagaggaaaaggggagcattttaatttcatggaacagcattccagtcca 2281 ggagcaaatgggctgcctcctccattataggatatactggaaggaacgggactccaactc 2341 ccagcctcagctctgtgaaattccctacagagtctcccaaaattcacatccaataaacag 2401 cctgcagccccgagtgacatatgtcctgtggatgacagctctgacagctgctggtgaaag 2461 ttcccacggaaatgagagggaattttgtctgcaaggtaaagccaattggatggcgtttgt 2521 ggcaccaagcatttgcattgctatcatcatggtgggcattttctcaacgcattacttcca 2581 gcaaaaggtgtttgttctcctagcagccctcagacctcagtggtgtagcagagaaattcc 2641 agatccagcaaatagcacttgcgctaagaaatatcccattgcagaggagaagacacagct 2701 gcccttggacaggctcctgatagactggcccacgcctgaagatcctgaaccgctggtcat 2761 cagtgaagtccttcatcaagtgaccccagttttcagacatcccccctgctccaactggcc 2821 acaaagggaaaaaggaatccaaggtcatcaggcctctgagaaagacatgatgcacagtgc 2881 ctcaagcccaccacctccaagagctctccaagctgagagcagacaactggtggatctgta 2941 caaggtgctggagagcaggggctccgacccaaagcccgaaaacccagcctgtccctggac 3001 ggtgctcccagcaggtgaccttcccacccatgatggctacttaccctccaacatagatga 3061 cctcccctcacatgaggcacctctcgctgactctctggaagaactggagcctcagcacat 3121 ctccctttctgttttcccctcaagttctcttcacccactcaccttctcctgtggtgataa 3181 gctgactctggatcagttaaagatgaggtgtgactccctcatgctctgagtggtgaggct 3241 tcaagccttaaagtcagtgtgccctcaaccagcacagcctgccccaattcccccagcccc 3301 tgctccagcagctgtcatctctgggtgccaccatcggtctggctgcagctagaggacagg 3361 caagccagctctgggggagtcttaggaactgggagttggtcttcactcagatgcctcatc 3421 ttgcctttcccagggccttaaaattacatccttcactgtgtggacctagagactccaact 3481 tgaattcctagtaactttcttggtatgctggccagaaagggaaatgaggaggagagtaga 3541 aaccacagctcttagtagtaatggcatacagtctagaggaccattcatgcaatgactatt 3601 tctaaagcacctgctacacagcaggctgtacacagcagatcagtactgttcaacagaact 3661 tcctgagatgatggaaatgttctacctctgcactcactgtccagtacattagacactagg 3721 cacattggctgttaatcacttggaatgtgtttagcttgactgaggaattaaattttgatt 3781 gtaaatttaaatcgccacacatggctagtggctactgtattggagtgcacagctctagat 3841 ggctcctagattattgagagccttcaaaacaaatcaacctagttctatagatgaagacat 3901 aaaagacactggtaaacaccaaggtaaaagggcccccaaggtggtcatgactggtctcat 3961 ttgcagaagtctaagaatgtacctttttctggccgggcgtggtagctcatgcctgtaatc 4021 ccagcactttgggaggctga HumanIL-12R2mRNAVariant2 (SEQIDNO:9) 1 tgcagagcacagagaaaggacatctgcgaggaaagttccctgatggctgtcaacaaagtg 61 ccacgtctctatggctgtgaacgctgagcacacgattttatcgcgcctatcatatcttgg 121 tgcataaacgcacctcacctcggtcaacccttgctccgtcttatgagacaggctttatta 181 tccgcattttatatgaggggaaactgacggtggagagagaattatcttgctcaaggcgac 241 acagcagagcccacaggtggcagaatcccacccgagcccgcttcgacccgcggggtggaa 301 accacgggcgcccgcccggctgcgcttccagagctgaactgagaagcgagtcctctccgc 361 cctgcggccaccgcccagccccgacccccgccccggcccgatcctcactcgccgccagct 421 ccccgcgcccaccccggagttggtggcgcagaggcgggaggcggaggcgggagggcgggc 481 gctggcaccgggaacgcccgagcgccggcagagagcgcggagagcgcgacacgtgcggcc 541 cagagcaccggggccacccggtccccgcaggcccgggaccgcgcccgctggcaggcgaca 601 cgtggtcacggtgatccatttgtaaagtcgggaataaatgacctctgaagtgttgtctgt 661 atattgatctgctaccagtaaaacatatctctgaagaatacggagttctataccagagtt 721 gattgttgatggcacatacttttagaggatgctcattggcatttatgtttataatcacgt 781 ggctgttgattaaagcaaaaatagatgcgtgcaagagaggcgatgtgactgtgaagcctt 841 cccatgtaattttacttggatccactgtcaatattacatgctctttgaagcccagacaag 901 gctgctttcactattccagacgtaacaagttaatcctgtacaagtttgacagaagaatca 961 attttcaccatggccactccctcaattctcaagtcacaggtcttccccttggtacaacct 1021 tgtttgtctgcaaactggcctgtatcaatagtgatgaaattcaaatatgtggagcagaga 1081 tcttcgttggtgttgctccagaacagcctcaaaatttatcctgcatacagaagggagaac 1141 aggggactgtggcctgcacctgggaaagaggacgagacacccacttatacactgagtata 1201 ctctacagctaagtggaccaaaaaatttaacctggcagaagcaatgtaaagacatttatt 1261 gtgactatttggactttggaatcaacctcacccctgaatcacctgaatccaatttcacag 1321 ccaaggttactgctgtcaatagtcttggaagctcctcttcacttccatccacattcacat 1381 tcttggacatagtgaggcctcttcctccgtgggacattagaatcaaatttcaaaaggctt 1441 ctgtgagcagatgtaccctttattggagagatgagggactggtactgcttaatcgactca 1501 gatatcggcccagtaacagcaggctctggaatatggttaatgttacaaaggccaaaggaa 1561 gacatgatttgctggatctgaaaccatttacagaatatgaatttcagatttcctctaagc 1621 tacatctttataagggaagttggagtgattggagtgaatcattgagagcacaaacaccag 1681 aagaagagcctactgggatgttagatgtctggtacatgaaacggcacattgactacagta 1741 gacaacagatttctcttttctggaagaatctgagtgtctcagaggcaagaggaaaaattc 1801 tccactatcaggtgaccttgcaggagctgacaggagggaaagccatgacacagaacatca 1861 caggacacacctcctggaccacagtcattcctagaaccggaaattgggctgtggctgtgt 1921 ctgcagcaaattcaaaaggcagttctctgcccactcgtattaacataatgaacctgtgtg 1981 aggcagggttgctggctcctcgccaggtctctgcaaactcagagggcatggacaacattc 2041 tggtgacttggcagcctcccaggaaagatccctctgctgttcaggagtacgtggtggaat 2101 ggagagagctccatccagggggtgacacacaggtccctctaaactggctacggagtcgac 2161 cctacaatgtgtctgctctgatttcagagaacataaaatcctacatctgttatgaaatcc 2221 gtgtgtatgcactctcaggggatcaaggaggatgcagctccatcctgggtaactctaagc 2281 acaaagcaccactgagtggcccccacattaatgccatcacagaggaaaaggggagcattt 2341 taatttcatggaacagcattccagtccaggagcaaatgggctgcctcctccattatagga 2401 tatactggaaggaacgggactccaactcccagcctcagctctgtgaaattccctacagag 2461 tctcccaaaattcacatccaataaacagcctgcagccccgagtgacatatgtcctgtgga 2521 tgacagctctgacagctgctggtgaaagttcccacggaaatgagagggaattttgtctgc 2581 aaggtaaagccaattggatggcgtttgtggcaccaagcatttgcattgctatcatcatgg 2641 tgggcattttctcaacgcattacttccagcaaaagagaagacacagctgcccttggacag 2701 gctcctgatagactggcccacgcctgaagatcctgaaccgctggtcatcagtgaagtcct 2761 tcatcaagtgaccccagttttcagacatcccccctgctccaactggccacaaagggaaaa 2821 aggaatccaaggtcatcaggcctctgagaaagacatgatgcacagtgcctcaagcccacc 2881 acctccaagagctctccaagctgagagcagacaactggtggatctgtacaaggtgctgga 2941 gagcaggggctccgacccaaagcccgaaaacccagcctgtccctggacggtgctcccagc 3001 aggtgaccttcccacccatgatggctacttaccctccaacatagatgacctcccctcaca 3061 tgaggcacctctcgctgactctctggaagaactggagcctcagcacatctccctttctgt 3121 tttcccctcaagttctcttcacccactcaccttctcctgtggtgataagctgactctgga 3181 tcagttaaagatgaggtgtgactccctcatgctctgagtggtgaggcttcaagccttaaa 3241 gtcagtgtgccctcaaccagcacagcctgccccaattcccccagcccctgctccagcagc 3301 tgtcatctctgggtgccaccatcggtctggctgcagctagaggacaggcaagccagctct 3361 gggggagtcttaggaactgggagttggtcttcactcagatgcctcatcttgcctttccca 3421 gggccttaaaattacatccttcactgtgtggacctagagactccaacttgaattcctagt 3481 aactttcttggtatgctggccagaaagggaaatgaggaggagagtagaaaccacagctct 3541 tagtagtaatggcatacagtctagaggaccattcatgcaatgactatttctaaagcacct 3601 gctacacagcaggctgtacacagcagatcagtactgttcaacagaacttcctgagatgat 3661 ggaaatgttctacctctgcactcactgtccagtacattagacactaggcacattggctgt 3721 taatcacttggaatgtgtttagcttgactgaggaattaaattttgattgtaaatttaaat 3781 cgccacacatggctagtggctactgtattggagtgcacagctctagatggctcctagatt 3841 attgagagccttcaaaacaaatcaacctagttctatagatgaagacataaaagacactgg 3901 taaacaccaaggtaaaagggcccccaaggtggtcatgactggtctcatttgcagaagtct 3961 aagaatgtacctttttctggccgggcgtggtagctcatgcctgtaatcccagcactttgg 4021 gaggctga HumanIL-12R2mRNAVariant3 (SEQIDNO:10) 1 tgcagagcacagagaaaggacatctgcgaggaaagttccctgatggctgtcaacaaagtg 61 ccacgtctctatggctgtgaacgctgagcacacgattttatcgcgcctatcatatcttgg 121 tgcataaacgcacctcacctcggtcaacccttgctccgtcttatgagacaggctttatta 181 tccgcattttatatgaggggaaactgacggtggagagagaattatcttgctcaaggcgac 241 acagcagagcccacaggtggcagaatcccacccgagcccgcttcgacccgcggggtggaa 301 accacgggcgcccgcccggctgcgcttccagagctgaactgagaagcgagtcctctccgc 361 cctgcggccaccgcccagccccgacccccgccccggcccgatcctcactcgccgccagct 421 ccccgcgcccaccccggagttggtggcgcagaggcgggaggcggaggcgggagggcgggc 481 gctggcaccgggaacgcccgagcgccggcagagagcgcggagagcgcgacacgtgcggcc 541 cagagcaccggggccacccggtccccgcaggcccgggaccgcgcccgctggcaggcgaca 601 cgtggaagaatacggagttctataccagagttgattgttgatggcacatacttttagagg 661 atgctcattggcatttatgtttataatcacgtggctgttgattaaagcaaaaatagatgc 721 gtgcaagagaggcgatgtgactgtgaagccttcccatgtaattttacttggatccactgt 781 caatattacatgctctttgaagcccagacaaggctgctttcactattccagacgtaacaa 841 gttaatcctgtacaagtttgacagaagaatcaattttcaccatggccactccctcaattc 901 tcaagtcacaggtcttccccttggtacaaccttgtttgtctgcaaactggcctgtatcaa 961 tagtgatgaaattcaaatatgtggagcagagatcttcgttggtgttgctccagaacagcc 1021 tcaaaatttatcctgcatacagaagggagaacaggggactgtggcctgcacctgggaaag 1081 aggacgagacacccacttatacactgagtatactctacagctaagtggaccaaaaaattt 1141 aacctggcagaagcaatgtaaagacatttattgtgactatttggactttggaatcaacct 1201 cacccctgaatcacctgaatccaatttcacagccaaggttactgctgtcaatagtcttgg 1261 aagctcctcttcacttccatccacattcacattcttggacatagtgaggcctcttcctcc 1321 gtgggacattagaatcaaatttcaaaaggcttctgtgagcagatgtaccctttattggag 1381 agatgagggactggtactgcttaatcgactcagatatcggcccagtaacagcaggctctg 1441 gaatatggttaatgttacaaaggccaaaggaagacatgatttgctggatctgaaaccatt 1501 tacagaatatgaatttcagatttcctctaagctacatctttataagggaagttggagtga 1561 ttggagtgaatcattgagagcacaaacaccagaagaagagcctactgggatgttagatgt 1621 ctggtacatgaaacggcacattgactacagtagacaacagatttctcttttctggaagaa 1681 tctgagtgtctcagaggcaagaggaaaaattctccactatcaggtgaccttgcaggagct 1741 gacaggagggaaagccatgacacagaacatcacaggacacacctcctggaccacagtcat 1801 tcctagaaccggaaattgggctgtggctgtgtctgcagcaaattcaaaaggcagttctct 1861 gcccactcgtattaacataatgaacctgtgtgaggcagggttgctggctcctcgccaggt 1921 ctctgcaaactcagagggcatggacaacattctggtgacttggcagcctcccaggaaaga 1981 tccctctgctgttcaggagtacgtggtggaatggagagagctccatccagggggtgacac 2041 acaggtccctctaaactggctacggagtcgaccctacaatgtgtctgctctgatttcaga 2101 aattccctacagagtctcccaaaattcacatccaataaacagcctgcagccccgagtgac 2161 atatgtcctgtggatgacagctctgacagctgctggtgaaagttcccacggaaatgagag 2221 ggaattttgtctgcaaggtaaagccaattggatggcgtttgtggcaccaagcatttgcat 2281 tgctatcatcatggtgggcattttctcaacgcattacttccagcaaaaggtgtttgttct 2341 cctagcagccctcagacctcagtggtgtagcagagaaattccagatccagcaaatagcac 2401 ttgcgctaagaaatatcccattgcagaggagaagacacagctgcccttggacaggctcct 2461 gatagactggcccacgcctgaagatcctgaaccgctggtcatcagtgaagtccttcatca 2521 agtgaccccagttttcagacatcccccctgctccaactggccacaaagggaaaaaggaat 2581 ccaaggtcatcaggcctctgagaaagacatgatgcacagtgcctcaagcccaccacctcc 2641 aagagctctccaagctgagagcagacaactggtggatctgtacaaggtgctggagagcag 2701 gggctccgacccaaagcccgaaaacccagcctgtccctggacggtgctcccagcaggtga 2761 ccttcccacccatgatggctacttaccctccaacatagatgacctcccctcacatgaggc 2821 acctctcgctgactctctggaagaactggagcctcagcacatctccctttctgttttccc 2881 ctcaagttctcttcacccactcaccttctcctgtggtgataagctgactctggatcagtt 2941 aaagatgaggtgtgactccctcatgctctgagtggtgaggcttcaagccttaaagtcagt 3001 gtgccctcaaccagcacagcctgccccaattcccccagcccctgctccagcagctgtcat 3061 ctctgggtgccaccatcggtctggctgcagctagaggacaggcaagccagctctggggga 3121 gtcttaggaactgggagttggtcttcactcagatgcctcatcttgcctttcccagggcct 3181 taaaattacatccttcactgtgtggacctagagactccaacttgaattcctagtaacttt 3241 cttggtatgctggccagaaagggaaatgaggaggagagtagaaaccacagctcttagtag 3301 taatggcatacagtctagaggaccattcatgcaatgactatttctaaagcacctgctaca 3361 cagcaggctgtacacagcagatcagtactgttcaacagaacttcctgagatgatggaaat 3421 gttctacctctgcactcactgtccagtacattagacactaggcacattggctgttaatca 3481 cttggaatgtgtttagcttgactgaggaattaaattttgattgtaaatttaaatcgccac 3541 acatggctagtggctactgtattggagtgcacagctctagatggctcctagattattgag 3601 agccttcaaaacaaatcaacctagttctatagatgaagacataaaagacactggtaaaca 3661 ccaaggtaaaagggcccccaaggtggtcatgactggtctcatttgcagaagtctaagaat 3721 gtacctttttctggccgggcgtggtagctcatgcctgtaatcccagcactttgggaggct 3781 ga HumanIL-12R2mRNAVariant4 (SEQIDNO:11) 1 tgcagagcacagagaaaggacatctgcgaggaaagttccctgatggctgtcaacaaagtg 61 ccacgtctctatggctgtgaacgctgagcacacgattttatcgcgcctatcatatcttgg 121 tgcataaacgcacctcacctcggtcaacccttgctccgtcttatgagacaggctttatta 181 tccgcattttatatgaggggaaactgacggtggagagagaattatcttgctcaaggcgac 241 acagcagagcccacaggtggcagaatcccacccgagcccgcttcgacccgcggggtggaa 301 accacgggcgcccgcccggctgcgcttccagagctgaactgagaagcgagtcctctccgc 361 cctgcggccaccgcccagccccgacccccgccccggcccgatcctcactcgccgccagct 421 ccccgcgcccaccccggagttggtggcgcagaggcgggaggcggaggcgggagggcgggc 481 gctggcaccgggaacgcccgagcgccggcagagagcgcggagagcgcgacacgtgcggcc 541 cagagcaccggggccacccggtccccgcaggcccgggaccgcgcccgctggcaggcgaca 601 cgtggaagaatacggagttctataccagagttgattgttgatggcacatacttttagagg 661 atgctcattggcatttatgtttataatcacgtggctgttgattaaagcaaaaatagatgc 721 gtgcaagagaggcgatgtgactgtgaagccttcccatgtaattttacttggatccactgt 781 caatattacatgctctttgaagcccagacaaggctgctttcactattccagacgtaacaa 841 gttaatcctgtacaagtttgacagaagaatcaattttcaccatggccactccctcaattc 901 tcaagtcacaggtcttccccttggtacaaccttgtttgtctgcaaactggcctgtatcaa 961 tagtgatgaaattcaaatatgtggagcagagatcttcgttggtgttgctccagaacagcc 1021 tcaaaatttatcctgcatacagaagggagaacaggggactgtggcctgcacctgggaaag 1081 aggacgagacacccacttatacactgagtatactctacagctaagtggaccaaaaaattt 1141 aacctggcagaagcaatgtaaagacatttattgtgactatttggactttggaatcaacct 1201 cacccctgaatcacctgaatccaatttcacagccaaggttactgctgtcaatagtcttgg 1261 aagctcctcttcacttccatccacattcacattcttggacatagtgaggcctcttcctcc 1321 gtgggacattagaatcaaatttcaaaaggcttctgtgagcagatgtaccctttattggag 1381 agatgagggactggtactgcttaatcgactcagatatcggcccagtaacagcaggctctg 1441 gaatatggttaatgttacaaaggccaaaggaagacatgatttgctggatctgaaaccatt 1501 tacagaatatgaatttcagatttcctctaagctacatctttataagggaagttggagtga 1561 ttggagtgaatcattgagagcacaaacaccagaagaagagcctactgggatgttagatgt 1621 ctggtacatgaaacggcacattgactacagtagacaacagatttctcttttctggaagaa 1681 tctgagtgtctcagaggcaagaggaaaaattctccactatcaggtgaccttgcaggagct 1741 gacaggagggaaagccatgacacagaacatcacaggacacacctcctggaccacagtcat 1801 tcctagaaccggaaattgggctgtggctgtgtctgcagcaaattcaaaaggcagttctct 1861 gcccactcgtattaacataatgaacctgtgtgaggcagggttgctggctcctcgccaggt 1921 ctctgcaaactcagagggcatggacaacattctggtgacttggcagcctcccaggaaaga 1981 tccctctgctgttcaggagtacgtggtggaatggagagagctccatccagggggtgacac 2041 acaggtccctctaaactggctacggagtcgaccctacaatgtgtctgctctgatttcaga 2101 gaacataaaatcctacatctgttatgaaatccgtgtgtatgcactctcaggggatcaagg 2161 aggatgcagctccatcctgggtaactctaagcacaaagcaccactgagtggcccccacat 2221 taatgccatcacagaggaaaaggggagcattttaatttcatggaacagcattccagtcca 2281 ggagcaaatgggctgcctcctccattataggatatactggaaggaacgggactccaactc 2341 ccagcctcagctctgtgaaattccctacagagtctcccaaaattcacatccaataaacag 2401 cctgcagccccgagtgacatatgtcctgtggatgacagctctgacagctgctggtgaaag 2461 ttcccacggaaatgagagggaattttgtctgcaaggagaagacacagctgcccttggaca 2521 ggctcctgatagactggcccacgcctgaagatcctgaaccgctggtcatcagtgaagtcc 2581 ttcatcaagtgaccccagttttcagacatcccccctgctccaactggccacaaagggaaa 2641 aaggaatccaaggtcatcaggcctctgagaaagacatgatgcacagtgcctcaagcccac 2701 cacctccaagagctctccaagctgagagcagacaactggtggatctgtacaaggtgctgg 2761 agagcaggggctccgacccaaagcccgaaaacccagcctgtccctggacggtgctcccag 2821 caggtgaccttcccacccatgatggctacttaccctccaacatagatgacctcccctcac 2881 atgaggcacctctcgctgactctctggaagaactggagcctcagcacatctccctttctg 2941 ttttcccctcaagttctcttcacccactcaccttctcctgtggtgataagctgactctgg 3001 atcagttaaagatgaggtgtgactccctcatgctctgagtggtgaggcttcaagccttaa 3061 agtcagtgtgccctcaaccagcacagcctgccccaattcccccagcccctgctccagcag 3121 ctgtcatctctgggtgccaccatcggtctggctgcagctagaggacaggcaagccagctc 3181 tgggggagtcttaggaactgggagttggtcttcactcagatgcctcatcttgcctttccc 3241 agggccttaaaattacatccttcactgtgtggacctagagactccaacttgaattcctag 3301 taactttcttggtatgctggccagaaagggaaatgaggaggagagtagaaaccacagctc 3361 ttagtagtaatggcatacagtctagaggaccattcatgcaatgactatttctaaagcacc 3421 tgctacacagcaggctgtacacagcagatcagtactgttcaacagaacttcctgagatga 3481 tggaaatgttctacctctgcactcactgtccagtacattagacactaggcacattggctg 3541 ttaatcacttggaatgtgtttagcttgactgaggaattaaattttgattgtaaatttaaa 3601 tcgccacacatggctagtggctactgtattggagtgcacagctctagatggctcctagat 3661 tattgagagccttcaaaacaaatcaacctagttctatagatgaagacataaaagacactg 3721 gtaaacaccaaggtaaaagggcccccaaggtggtcatgactggtctcatttgcagaagtc 3781 taagaatgtacctttttctggccgggcgtggtagctcatgcctgtaatcccagcactttg 3841 ggaggctga HumanIL-23RmRNA (SEQIDNO:12) 1 acaagggtggcagcctggctctgaagtggaattatgtgcttcaaacaggttgaaagaggg 61 aaacagtcttttcctgcttccagacatgaatcaggtcactattcaatgggatgcagtaat 121 agccctttacatactcttcagctggtgtcatggaggaattacaaatataaactgctctgg 181 ccacatctgggtagaaccagccacaatttttaagatgggtatgaatatctctatatattg 241 ccaagcagcaattaagaactgccaaccaaggaaacttcatttttataaaaatggcatcaa 301 agaaagatttcaaatcacaaggattaataaaacaacagctcggctttggtataaaaactt 361 tctggaaccacatgcttctatgtactgcactgctgaatgtcccaaacattttcaagagac 421 actgatatgtggaaaagacatttcttctggatatccgccagatattcctgatgaagtaac 481 ctgtgtcatttatgaatattcaggcaacatgacttgcacctggaatgctgggaagctcac 541 ctacatagacacaaaatacgtggtacatgtgaagagtttagagacagaagaagagcaaca 601 gtatctcacctcaagctatattaacatctccactgattcattacaaggtggcaagaagta 661 cttggtttgggtccaagcagcaaacgcactaggcatggaagagtcaaaacaactgcaaat 721 tcacctggatgatatagtgataccttctgcagccgtcatttccagggctgagactataaa 781 tgctacagtgcccaagaccataatttattgggatagtcaaacaacaattgaaaaggtttc 841 ctgtgaaatgagatacaaggctacaacaaaccaaacttggaatgttaaagaatttgacac 901 caattttacatatgtgcaacagtcagaattctacttggagccaaacattaagtacgtatt 961 tcaagtgagatgtcaagaaacaggcaaaaggtactggcagccttggagttcaccgttttt 1021 tcataaaacacctgaaacagttccccaggtcacatcaaaagcattccaacatgacacatg 1081 gaattctgggctaacagttgcttccatctctacagggcaccttacttctgacaacagagg 1141 agacattggacttttattgggaatgatcgtctttgctgttatgttgtcaattctttcttt 1201 gattgggatatttaacagatcattccgaactgggattaaaagaaggatcttattgttaat 1261 accaaagtggctttatgaagatattcctaatatgaaaaacagcaatgttgtgaaaatgct 1321 acaggaaaatagtgaacttatgaataataattccagtgagcaggtcctatatgttgatcc 1381 catgattacagagataaaagaaatcttcatcccagaacacaagcctacagactacaagaa 1441 ggagaatacaggacccctggagacaagagactacccgcaaaactcgctattcgacaatac 1501 tacagttgtatatattcctgatctcaacactggatataaaccccaaatttcaaattttct 1561 gcctgagggaagccatctcagcaataataatgaaattacttccttaacacttaaaccacc 1621 agttgattccttagactcaggaaataatcccaggttacaaaagcatcctaattttgcttt 1681 ttctgtttcaagtgtgaattcactaagcaacacaatatttcttggagaattaagcctcat 1741 attaaatcaaggagaatgcagttctcctgacatacaaaactcagtagaggaggaaaccac 1801 catgcttttggaaaatgattcacccagtgaaactattccagaacagaccctgcttcctga 1861 tgaatttgtctcctgtttggggatcgtgaatgaggagttgccatctattaatacttattt 1921 tccacaaaatattttggaaagccacttcaataggatttcactcttggaaaagtagagctg 1981 tgtggtcaaaatcaatatgagaaagctgccttgcaatctgaacttgggttttccctgcaa 2041 tagaaattgaattctgcctctttttgaaaaaaatgtattcacatacaaatcttcacatgg 2101 acacatgttttcatttcccttggataaatacctaggtaggggattgctgggccatatgat 2161 aagcatatgtttcagttctaccaatcttgtttccagagtagtgacatttctgtgctccta 2221 ccatcaccatgtaagaattcccgggagctccatgcctttttaattttagccattcttctg 2281 cctcatttcttaaaattagagaattaaggtcccgaaggtggaacatgcttcatggtcaca 2341 catacaggcacaaaaacagcattatgtggacgcctcatgtattttttatagagtcaacta 2401 tttcctctttattttccctcattgaaagatgcaaaacagctctctattgtgtacagaaag 2461 ggtaaataatgcaaaatacctggtagtaaaataaatgctgaaaattttcctttaaaatag 2521 aatcattaggccaggcgtggtggctcatgcttgtaatcccagcactttggtaggctgagg 2581 tgggtggatcacctgaggtcaggagttcgagtccagcctggccaatatgctgaaaccctg 2641 tctctactaaaattacaaaaattagccggccatggtggcaggtgcttgtaatcccagcta 2701 cttgggaggctgaggcaggagaatcacttgaaccaggaaggcagaggttgcactgagctg 2761 agattgtgccactgcactccagcctgggcaacaagagcaaaactctgtctggaaaaaaaa 2821 aaaaaa

[0277] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0278] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein can be designed using the software available at the Integrated DNA Technologies website.

[0279] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0280] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an anti sense orientation to a target nucleic acid of interest).

[0281] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0282] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, -units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987). Non-limiting examples of antisense nucleic acids are described in Vaknin-Dembinsky et al., J. Immunol. 176(12): 7768-7774, 2006.

[0283] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein (e.g., specificity for an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA, e.g., specificity for any one of SEQ ID NOs: 1-12). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA can be designed based upon the nucleotide sequence of any of the IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, and IL-23R mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0284] An inhibitor nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0285] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Natl. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0286] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0287] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0288] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Natl. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0289] Another means by which expression of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R protein) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0290] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0291] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0292] The siRNA molecules used to decrease expression of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4 or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0293] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 1-12, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0294] Non-limiting examples of siRNAs targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R are described in Tan et al., J. Alzheimers Dis. 38(3): 633-646, 2014; Niimi et al., J. Neuroimmunol. 254(1-2): 39-45, 2013. Non-limiting examples of short hairpin RNA (shRNA) targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R are described in Bak et al., BMC Dermatol. 11:5, 2011.

[0295] Non-limiting examples of inhibitory nucleic acids are microRNAs (e.g., microRNA-29 (Brain et al., Immunity 39(3):521-536, 2013), miR-10a (Xue et al., J. Immunol. 187(11):5879-5886, 2011), microRNA-155 (Podsiad et al., Am. J. Physiol. Lung Cell Mol. Physiol. 310(5):L465-75, 2016).

[0296] In some embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R can be administered to a subject (e.g., a human subject) in need thereof.

[0297] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3 end of DNA or RNA.

[0298] Any of the inhibitor nucleic acids described herein can be formulated for administration to the gastrointestinal tract. See, e.g., the formulation methods described in US 2016/0090598 and Schoellhammer et al., Gastroenterology, doi: 10.1053/j.gastro.2017.01.002, 2017.

[0299] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0300] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0301] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R) with a Tm of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive),

[0302] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0303] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0304] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0305] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0306] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0307] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0308] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0309] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0310] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0311] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0312] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0313] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0314] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0315] In some embodiments, the IL-12/IL-23 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 1, IL-12R 2, or IL-23R, or a combination thereof.

[0316] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv).sub.2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv.sub.2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0317] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab)2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0318] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0319] In some embodiments, the antibody is ustekinumab (CNTO 1275, Stelara) or a variant thereof (Krueger et al., N. Engl. J. Med. 356(6):580-592, 2007; Kauffman et al., J. Invest. Dermatol. 123(6):1037-1044, 2004; Gottlieb et al., Curr. Med. Res. Opin. 23(5):1081-1092, 2007; Leonardi et al., Lancet 371(9625):1665-1674, 2008; Papp et al., Lancet 371(9625):1675-1684, 2008). In some embodiments, the antibody is briakinumab (ABT-874, J-695) or a variant thereof (Gordon et al., J. Invest. Dermatol. 132(2):304-314, 2012; Kimball et al., Arch Dermatol. 144(2): 200-207, 2008).

[0320] In some embodiments, the antibody is guselkumab (CNTO-1959) (Callis-Duffin et al., J. Am. Acad. Dermatol. 70(5 Suppl 1), 2014); AB162 (Sofen et al., J. Allergy Clin. Immunol. 133: 1032-40, 2014); tildrakizumab (MK-3222, SCH900222) (Papp et al. (2015) Br. J. Dermatol. 2015); Langley et al., Oral Presentation at: American Academy of Dermatology, March 21-25, Denver Colo., 2014); AMG 139 (MEDI2070, brazikumab) (Gomollon, Gastroenterol. Hepatol. 38(Suppl. 1):13-19, 2015; Kock et al., Br. J. Pharmacol. 172(1):159-172, 2015); FM-202 (Tang et al., Immunology 135(2):112-124, 2012); FM-303 (Tang et al., Immunology 135(2):112-124, 2012); ADC-1012 (Tang et al., Immunology 135(2):112-124, 2012); LY-2525623 (Gaffen et al., Nat. Rev. Immunol. 14:585-600, 2014; Sands, Gastroenterol. Hepatol. 12(12):784-786, 2016), LY-3074828 (Coskun et al., Trends Pharmacol. Sci. 38(2):127-142, 2017), BI-655066 (risankizumab) (Singh et al., MAbs 7(4):778-791, 2015; Krueger et al., J. Allergy Clin. Immunol. 136(1):116-124, 2015) or a variant thereof.

[0321] See e.g., Tang et al., Immunology 135(2):112-124, 2012. Further teachings of IL-12/IL-23 antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 6,902,734; 7,247,711; 7,252,971; and 7,491,391; US 2012/0288494; and US 2013/0302343, each of which is incorporated by reference in its entirety.

[0322] In some embodiments, the IL-12/IL-23 inhibitor is PTG-200, an IL-23R inhibitor currently in preclinical development by Protagonist Therapeutics.

[0323] In some embodiments, the IL-12/IL-23 inhibitor is Mirikizumab (LY 3074828), an IL-23R inhibitor currently in clinical development (Phase II) by Eli Lilly. In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5M (e.g., less than 0.510.sup.5M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7M, less than 0.510.sup.7M, less than 110.sup.8M, less than 0.510.sup.8M, less than 110.sup.9M, less than 0.510.sup.9M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11M, less than 0.510.sup.11M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0324] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.9 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11M, or about 0.510.sup.11M (inclusive); about 0.510.sup.11M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11M (inclusive); about 110.sup.11M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9 M, or about 0.510.sup.9M (inclusive); about 0.510.sup.9M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8 M, about 0.510.sup.8M, or about 110.sup.9M (inclusive); about 110.sup.9M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7 M, about 110.sup.8M, or about 0.510.sup.8M (inclusive); about 0.510.sup.8M to about 110.sup.5 M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5M to about 110.sup.5M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0325] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0326] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, s about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1 about 110.sup.5 M.sup.1s.sup.1 or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins

[0327] In some embodiments, the IL-12/IL-23 inhibitor is a fusion protein, a soluble antagonist, or an antimicrobial peptide. In some embodiments, the fusion protein comprises a soluble fragment of a receptor of IL-12 or a soluble fragment of a receptor of IL-23. In some embodiments, the fusion protein comprises an extracellular domain of a receptor of IL-12 or an extracellular domain of a receptor of IL-23.

[0328] In some embodiments, the fusion protein is adnectin or a variant thereof (Tang et al., Immunology 135(2):112-124, 2012). In some embodiments, the soluble antagonist is a human IL-23Ra-chain mRNA transcript (Raymond et al., J. Immunol. 185(12):7302-7308, 2010). In some embodiments, the IL-12/IL-23 is an antimicrobial peptide (e.g., MP-196 (Wenzel et al., PNAS 111(14):E1409-E1418, 2014)).

Small Molecules

[0329] In some embodiments, the IL-12/IL-23 inhibitor is a small molecule. In some embodiments, the small molecule is STA-5326 (apilimod) or a variant thereof (Keino et al., Arthritis Res. Ther. 10: R122, 2008; Wada et al., Blood 109(3):1156-1164, 2007; Sands et al., Inflamm. Bowel Dis. 16(7):1209-1218, 2010).

TNF Inhibitors

[0330] The term TNF inhibitor refers to an agent which directly or indirectly inhibits, impairs, reduces, down-regulates, or blocks TNF activity and/or expression. In some embodiments, a TNF inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, a soluble TNF receptor (a soluble TNFR1 or a soluble TNFR2), or a small molecule TNF antagonist. In some embodiments, the inhibitory nucleic acid is a ribozyme, small hairpin RNA, a small interfering RNA, an antisense nucleic acid, or an aptamer.

[0331] Exemplary TNF inhibitors that directly inhibit, impair, reduce, down-regulate, or block TNF activity and/or expression can, e.g., inhibit or reduce binding of TNF to its receptor (TNFR1 and/or TNFR2) and/or inhibit or decrease the expression level of TNF or a receptor of TNF (TNFR1 or TNFR2) in a cell (e.g., a mammalian cell). Non-limiting examples of TNF inhibitors that directly inhibit, impair, reduce, down-regulate, or block TNF activity and/or expression include inhibitory nucleic acids (e.g., any of the examples of inhibitory nucleic acids described herein), an antibody or fragment thereof, a fusion protein, a soluble TNF receptor (e.g., a soluble TNFR1 or soluble TNFR2), and a small molecule TNF antagonist.

[0332] Exemplary TNF inhibitors that can indirectly inhibit, impair, reduce, down-regulate, or block TNF activity and/or expression can, e.g., inhibit or decrease the level of downstream signaling of a TNF receptor (e.g., TNFR1 or TNFR2) in a mammalian cell (e.g., decrease the level and/or activity of one or more of the following signaling proteins: TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, and NF-B in a mammalian cell), and/or decrease the level of TNF-induced gene expression in a mammalian cell (e.g., decrease the transcription of genes regulated by, e.g., one or more transcription factors selected from the group of NF-B, c-Jun, and ATF2). A description of downstream signaling of a TNF receptor is provided in Wajant et al., Cell Death Differentiation 10:45-65, 2003 (incorporated herein by reference). For example, such indirect TNF inhibitors can be an inhibitory nucleic acid that targets (decreases the expression) a signaling component downstream of a TNF receptor (e.g., any one or more of the signaling components downstream of a TNF receptor described herein or known in the art), a TNF-induced gene (e.g., any TNF-induced gene known in the art), or a transcription factor selected from the group of NF-B, c-Jun, and ATF2.

[0333] In other examples, such indirect TNF inhibitors can be a small molecule inhibitor of a signaling component downstream of a TNF receptor (e.g., any of the signaling components downstream of a TNF receptor described herein or known in the art), a small molecule inhibitor of a protein encoded by a TNF-induced gene (e.g., any protein encoded by a TNF-induced gene known in the art), and a small molecule inhibitor of a transcription factor selected from the group of NF-B, c-Jun, and ATF2.

[0334] In other embodiments, TNF inhibitors that can indirectly inhibit, impair, reduce, down-regulate, or block one or more components in a mammalian cell (e.g., a macrophage, a CD4+ lymphocyte, a NK cell, a neutrophil, a mast cell, a eosinophil, or a neuron) that are involved in the signaling pathway that results in TNF mRNA transcription, TNF mRNA stabilization, and TNF mRNA translation (e.g., one or more components selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, and MK2). For example, such indirect TNF inhibitors can be an inhibitory nucleic acid that targets (decreases the expression) of a component in a mammalian cell that is involved in the signaling pathway that results in TNF mRNA transcription, TNF mRNA stabilization, and TNF mRNA translation (e.g., a component selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, and MK2). In other examples, an indirect TNF inhibitors is a small molecule inhibitor of a component in a mammalian cell that is involved in the signaling pathway that results in TNF mRNA transcription, TNF mRNA stabilization, and TNF mRNA translation (e.g., a component selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, and MK2).

Inhibitory Nucleic Acids

[0335] Inhibitory nucleic acids that can decrease the expression of TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 13-49).

TABLE-US-00002 HumanTNFCDS (SEQIDNO:13) ATGAGCACTGAAAGCATGATCCGGGACGTGGAGCTGGCCGAGGAGGCGCTCCCCAAGAAGACAGGGGGGCCCCAGGGCTCCA GGCGGTGCTTGTTCCTCAGCCTCTTCTCCTTCCTGATCGTGGCAGGCGCCACCACGCTCTTCTGCCTGCTGCACTTTGGAGT GATCGGCCCCCAGAGGGAAGAGTTCCCCAGGGACCTCTCTCTAATCAGCCCTCTGGCCCAGGCAGTCAGATCATCTTCTCGA ACCCCGAGTGACAAGCCTGTAGCCCATGTTGTAGCAAACCCTCAAGCTGAGGGGCAGCTCCAGTGGCTGAACCGCCGGGCCA ATGCCCTCCTGGCCAATGGCGTGGAGCTGAGAGATAACCAGCTGGTGGTGCCATCAGAGGGCCTGTACCTCATCTACTCCCA GGTCCTCTTCAAGGGCCAAGGCTGCCCCTCCACCCATGTGCTCCTCACCCACACCATCAGCCGCATCGCCGTCTCCTACCAG ACCAAGGTCAACCTCCTCTCTGCCATCAAGAGCCCCTGCCAGAGGGAGACCCCAGAGGGGGCTGAGGCCAAGCCCTGGTATG AGCCCATCTATCTGGGAGGGGTCTTCCAGCTGGAGAAGGGTGACCGACTCAGCGCTGAGATCAATCGGCCCGACTATCTCGA CTTTGCCGAGTCTGGGCAGGTCTACTTTGGGATCATTGCCCTGTGA HumanTNFR1CDS (SEQIDNO:14) ATGGGCCTCTCCACCGTGCCTGACCTGCTGCTGCCGCTGGTGCTCCTGGAGCTGTTGGTGGGAATATACCCCTCAGGGGTTA TTGGACTGGTCCCTCACCTAGGGGACAGGGAGAAGAGAGATAGTGTGTGTCCCCAAGGAAAATATATCCACCCTCAAAATAA TTCGATTTGCTGTACCAAGTGCCACAAAGGAACCTACTTGTACAATGACTGTCCAGGCCCGGGGCAGGATACGGACTGCAGG GAGTGTGAGAGCGGCTCCTTCACCGCTTCAGAAAACCACCTCAGACACTGCCTCAGCTGCTCCAAATGCCGAAAGGAAATGG GTCAGGTGGAGATCTCTTCTTGCACAGTGGACCGGGACACCGTGTGTGGCTGCAGGAAGAACCAGTACCGGCATTATTGGAG TGAAAACCTTTTCCAGTGCTTCAATTGCAGCCTCTGCCTCAATGGGACCGTGCACCTCTCCTGCCAGGAGAAACAGAACACC GTGTGCACCTGCCATGCAGGTTTCTTTCTAAGAGAAAACGAGTGTGTCTCCTGTAGTAACTGTAAGAAAAGCCTGGAGTGCA CGAAGTTGTGCCTACCCCAGATTGAGAATGTTAAGGGCACTGAGGACTCAGGCACCACAGTGCTGTTGCCCCTGGTCATTTT CTTTGGTCTTTGCCTTTTATCCCTCCTCTTCATTGGTTTAATGTATCGCTACCAACGGTGGAAGTCCAAGCTCTACTCCATT GTTTGTGGGAAATCGACACCTGAAAAAGAGGGGGAGCTTGAAGGAACTACTACTAAGCCCCTGGCCCCAAACCCAAGCTTCA GTCCCACTCCAGGCTTCACCCCCACCCTGGGCTTCAGTCCCGTGCCCAGTTCCACCTTCACCTCCAGCTCCACCTATACCCC CGGTGACTGTCCCAACTTTGCGGCTCCCCGCAGAGAGGTGGCACCACCCTATCAGGGGGCTGACCCCATCCTTGCGACAGCC CTCGCCTCCGACCCCATCCCCAACCCCCTTCAGAAGTGGGAGGACAGCGCCCACAAGCCACAGAGCCTAGACACTGATGACC CCGCGACGCTGTACGCCGTGGTGGAGAACGTGCCCCCGTTGCGCTGGAAGGAATTCGTGCGGCGCCTAGGGCTGAGCGACCA CGAGATCGATCGGCTGGAGCTGCAGAACGGGCGCTGCCTGCGCGAGGCGCAATACAGCATGCTGGCGACCTGGAGGCGGCGC ACGCCGCGGCGCGAGGCCACGCTGGAGCTGCTGGGACGCGTGCTCCGCGACATGGACCTGCTGGGCTGCCTGGAGGACATCG AGGAGGCGCTTTGCGGCCCCGCCGCCCTCCCGCCCGCGCCCAGTCTTCTCAGATGA HumanTNFR2CDS (SEQIDNO:15) ATGGCGCCCGTCGCCGTCTGGGCCGCGCTGGCCGTCGGACTGGAGCTCTGGGCTGCGGCGCACGCCTTGCCCGCCCAGGTGG CATTTACACCCTACGCCCCGGAGCCCGGGAGCACATGCCGGCTCAGAGAATACTATGACCAGACAGCTCAGATGTGCTGCAG CAAATGCTCGCCGGGCCAACATGCAAAAGTCTTCTGTACCAAGACCTCGGACACCGTGTGTGACTCCTGTGAGGACAGCACA TACACCCAGCTCTGGAACTGGGTTCCCGAGTGCTTGAGCTGTGGCTCCCGCTGTAGCTCTGACCAGGTGGAAACTCAAGCCT GCACTCGGGAACAGAACCGCATCTGCACCTGCAGGCCCGGCTGGTACTGCGCGCTGAGCAAGCAGGAGGGGTGCCGGCTGTG CGCGCCGCTGCGCAAGTGCCGCCCGGGCTTCGGCGTGGCCAGACCAGGAACTGAAACATCAGACGTGGTGTGCAAGCCCTGT GCCCCGGGGACGTTCTCCAACACGACTTCATCCACGGATATTTGCAGGCCCCACCAGATCTGTAACGTGGTGGCCATCCCTG GGAATGCAAGCATGGATGCAGTCTGCACGTCCACGTCCCCCACCCGGAGTATGGCCCCAGGGGCAGTACACTTACCCCAGCC AGTGTCCACACGATCCCAACACACGCAGCCAACTCCAGAACCCAGCACTGCTCCAAGCACCTCCTTCCTGCTCCCAATGGGC CCCAGCCCCCCAGCTGAAGGGAGCACTGGCGACTTCGCTCTTCCAGTTGGACTGATTGTGGGTGTGACAGCCTTGGGTCTAC TAATAATAGGAGTGGTGAACTGTGTCATCATGACCCAGGTGAAAAAGAAGCCCTTGTGCCTGCAGAGAGAAGCCAAGGTGCC TCACTTGCCTGCCGATAAGGCCCGGGGTACACAGGGCCCCGAGCAGCAGCACCTGCTGATCACAGCGCCGAGCTCCAGCAGC AGCTCCCTGGAGAGCTCGGCCAGTGCGTTGGACAGAAGGGCGCCCACTCGGAACCAGCCACAGGCACCAGGCGTGGAGGCCA GTGGGGCCGGGGAGGCCCGGGCCAGCACCGGGAGCTCAGATTCTTCCCCTGGTGGCCATGGGACCCAGGTCAATGTCACCTG CATCGTGAACGTCTGTAGCAGCTCTGACCACAGCTCACAGTGCTCCTCCCAAGCCAGCTCCACAATGGGAGACACAGATTCC AGCCCCTCGGAGTCCCCGAAGGACGAGCAGGTCCCCTTCTCCAAGGAGGAATGTGCCTTTCGGTCACAGCTGGAGACGCCAG AGACCCTGCTGGGGAGCACCGAAGAGAAGCCCCTGCCCCTTGGAGTGCCTGATGCTGGGATGAAGCCCAGTTAA HumanTRADDCDS (SEQIDNO:16) ATGGCAGCTGGGCAAAATGGGCACGAAGAGTGGGTGGGCAGCGCATACCTGTTTGTGGAGTCCTCGCTGGACAAGGTGGTCC TGTCGGATGCCTACGCGCACCCCCAGCAGAAGGTGGCAGTGTACAGGGCTCTGCAGGCTGCCTTGGCAGAGAGCGGCGGGAG CCCGGACGTGCTGCAGATGCTGAAGATCCACCGCAGCGACCCGCAGCTGATCGTGCAGCTGCGATTCTGCGGGCGGCAGCCC TGTGGCCGCTTCCTCCGCGCCTACCGCGAGGGGGCGCTGCGCGCCGCGCTGCAGAGGAGCCTGGCGGCCGCGCTCGCCCAGC ACTCGGTGCCGCTGCAACTGGAGCTGCGCGCCGGCGCCGAGCGGCTGGACGCTTTGCTGGCGGACGAGGAGCGCTGTTTGAG TTGCATCCTAGCCCAGCAGCCCGACCGGCTCCGGGATGAAGAACTGGCTGAGCTGGAGGATGCGCTGCGAAATCTGAAGTGC GGCTCGGGGGCCCGGGGTGGCGACGGGGAGGTCGCTTCGGCCCCCTTGCAGCCCCCGGTGCCCTCTCTGTCGGAGGTGAAGC CGCCGCCGCCGCCGCCACCTGCCCAGACTTTTCTGTTCCAGGGTCAGCCTGTAGTGAATCGGCCGCTGAGCCTGAAGGACCA ACAGACGTTCGCGCGCTCTGTGGGTCTCAAATGGCGCAAGGTGGGGCGCTCACTGCAGCGAGGCTGCCGGGCGCTGCGGGAC CCGGCGCTGGACTCGCTGGCCTACGAGTACGAGCGCGAGGGACTGTACGAGCAGGCCTTCCAGCTGCTGCGGCGCTTCGTGC AGGCCGAGGGCCGCCGCGCCACGCTGCAGCGCCTGGTGGAGGCACTCGAGGAGAACGAGCTCACCAGCCTGGCAGAGGACTT GCTGGGCCTGACCGATCCCAATGGCGGCCTGGCCTAG HumanTRAF2CDS (SEQIDNO:17) ATGGCTGCAGCTAGCGTGACCCCCCCTGGCTCCCTGGAGTTGCTACAGCCCGGCTTCTCCAAGACCCTCCTGGGGACCAAGC TGGAAGCCAAGTACCTGTGCTCCGCCTGCAGAAACGTCCTCCGCAGGCCCTTCCAGGCGCAGTGTGGCCACCGGTACTGCTC CTTCTGCCTGGCCAGCATCCTCAGCTCTGGGCCTCAGAACTGTGCTGCCTGTGTTCACGAGGGCATATATGAAGAAGGCATT TCTATTTTAGAAAGCAGTTCGGCCTTCCCAGATAATGCTGCCCGCAGGGAGGTGGAGAGCCTGCCGGCCGTCTGTCCCAGTG ATGGATGCACCTGGAAGGGGACCCTGAAAGAATACGAGAGCTGCCACGAAGGCCGCTGCCCGCTCATGCTGACCGAATGTCC CGCGTGCAAAGGCCTGGTCCGCCTTGGTGAAAAGGAGCGCCACCTGGAGCACGAGTGCCCGGAGAGAAGCCTGAGCTGCCGG CATTGCCGGGCACCCTGCTGCGGAGCAGACGTGAAGGCGCACCACGAGGTCTGCCCCAAGTTCCCCTTAACTTGTGACGGCT GCGGCAAGAAGAAGATCCCCCGGGAGAAGTTTCAGGACCACGTCAAGACTTGTGGCAAGTGTCGAGTCCCTTGCAGATTCCA CGCCATCGGCTGCCTCGAGACGGTAGAGGGTGAGAAACAGCAGGAGCACGAGGTGCAGTGGCTGCGGGAGCACCTGGCCATG CTACTGAGCTCGGTGCTGGAGGCAAAGCCCCTCTTGGGAGACCAGAGCCACGCGGGGTCAGAGCTCCTGCAGAGGTGCGAGA GCCTGGAGAAGAAGACGGCCACTTTTGAGAACATTGTCTGCGTCCTGAACCGGGAGGTGGAGAGGGTGGCCATGACTGCCGA GGCCTGCAGCCGGCAGCACCGGCTGGACCAAGACAAGATTGAAGCCCTGAGTAGCAAGGTGCAGCAGCTGGAGAGGAGCATT GGCCTCAAGGACCTGGCGATGGCTGACTTGGAGCAGAAGGTCTTGGAGATGGAGGCATCCACCTACGATGGGGTCTTCATCT GGAAGATCTCAGACTTCGCCAGGAAGCGCCAGGAAGCTGTGGCTGGCCGCATACCCGCCATCTTCTCCCCAGCCTTCTACAC CAGCAGGTACGGCTACAAGATGTGTCTGCGTATCTACCTGAACGGCGACGGCACCGGGCGAGGAACACACCTGTCCCTCTTC TTTGTGGTGATGAAGGGCCCGAATGACGCCCTGCTGCGGTGGCCCTTCAACCAGAAGGTGACCTTAATGCTGCTCGACCAGA ATAACCGGGAGCACGTGATTGACGCCTTCAGGCCCGACGTGACTTCATCCTCTTTTCAGAGGCCAGTCAACGACATGAACAT CGCAAGCGGCTGCCCCCTCTTCTGCCCCGTCTCCAAGATGGAGGCAAAGAATTCCTACGTGCGGGACGATGCCATCTTCATC AAGGCCATTGTGGACCTGACAGGGCTCTAA HumanMEKK1CDS (SEQIDNO:18) ATGGCGGCGGCGGCGGGGAATCGCGCCTCGTCGTCGGGATTCCCGGGCGCCAGGGCTACGAGCCCTGAGGCAGGCGGCGGCG GAGGAGCCCTCAAGGCGAGCAGCGCGCCCGCGGCTGCCGCGGGACTGCTGCGGGAGGCGGGCAGCGGGGGCCGCGAGCGGGC GGACTGGCGGCGGCGGCAGCTGCGCAAAGTGCGGAGTGTGGAGCTGGACCAGCTGCCTGAGCAGCCGCTCTTCCTTGCCGCC TCACCGCCGGCCTCCTCGACTTCCCCGTCGCCGGAGCCCGCGGACGCAGCGGGGAGTGGGACCGGCTTCCAGCCTGTGGCGG TGCCGCCGCCCCACGGAGCCGCGAGCCGCGGCGGCGCCCACCTTACCGAGTCGGTGGCGGCGCCGGACAGCGGCGCCTCGAG TCCCGCAGCGGCCGAGCCCGGGGAGAAGCGGGCGCCCGCCGCCGAGCCGTCTCCTGCAGCGGCCCCCGCCGGTCGTGAGATG GAGAATAAAGAAACTCTCAAAGGGTTGCACAAGATGGATGATCGTCCAGAGGAACGAATGATCAGGGAGAAACTGAAGGCAA CCTGTATGCCAGCCTGGAAGCACGAATGGTTGGAAAGGAGAAATAGGCGAGGGCCTGTGGTGGTAAAACCAATCCCAGTTAA AGGAGATGGATCTGAAATGAATCACTTAGCAGCTGAGTCTCCAGGAGAGGTCCAGGCAAGTGCGGCTTCACCAGCTTCCAAA GGCCGACGCAGTCCTTCTCCTGGCAACTCCCCATCAGGTCGCACAGTGAAATCAGAATCTCCAGGAGTAAGGAGAAAAAGAG TTTCCCCAGTGCCTTTTCAGAGTGGCAGAATCACACCACCCCGAAGAGCCCCTTCACCAGATGGCTTCTCACCATATAGCCC TGAGGAAACAAACCGCCGTGTTAACAAAGTGATGCGGGCCAGACTGTACTTACTGCAGCAGATAGGGCCTAACTCTTTCCTG ATTGGAGGAGACAGCCCAGACAATAAATACCGGGTGTTTATTGGGCCTCAGAACTGCAGCTGTGCACGTGGAACATTCTGTA TTCATCTGCTATTTGTGATGCTCCGGGTGTTTCAACTAGAACCTTCAGACCCAATGTTATGGAGAAAAACTTTAAAGAATTT TGAGGTTGAGAGTTTGTTCCAGAAATATCACAGTAGGCGTAGCTCAAGGATCAAAGCTCCATCTCGTAACACCATCCAGAAG TTTGTTTCACGCATGTCAAATTCTCATACATTGTCATCATCTAGTACTTCTACGTCTAGTTCAGAAAACAGCATAAAGGATG AAGAGGAACAGATGTGTCCTATTTGCTTGTTGGGCATGCTTGATGAAGAAAGTCTTACAGTGTGTGAAGACGGCTGCAGGAA CAAGCTGCACCACCACTGCATGTCAATTTGGGCAGAAGAGTGTAGAAGAAATAGAGAACCTTTAATATGTCCCCTTTGTAGA TCTAAGTGGAGATCTCATGATTTCTACAGCCACGAGTTGTCAAGTCCTGTGGATTCCCCTTCTTCCCTCAGAGCTGCACAGC AGCAAACCGTACAGCAGCAGCCTTTGGCTGGATCACGAAGGAATCAAGAGAGCAATTTTAACCTTACTCATTATGGAACTCA GCAAATCCCTCCTGCTTACAAAGATTTAGCTGAGCCATGGATTCAGGTGTTTGGAATGGAACTCGTTGGCTGCTTATTTTCT AGAAACTGGAATGTGAGAGAGATGGCCCTCAGGCGTCTTTCCCATGATGTCAGTGGGGCCCTGCTGTTGGCAAATGGGGAGA GCACTGGAAATTCTGGGGGCAGCAGTGGAAGCAGCCCGAGTGGGGGAGCCACCAGTGGGTCTTCCCAGACCAGTATCTCAGG AGATGTGGTGGAGGCATGCTGCAGCGTTCTGTCAATGGTCTGTGCTGACCCTGTCTACAAAGTGTACGTTGCTGCTTTAAAA ACATTGAGAGCCATGCTGGTATATACTCCTTGCCACAGTTTAGCGGAAAGAATCAAACTTCAGAGACTTCTCCAGCCAGTTG TAGACACCATCCTAGTCAAATGTGCAGATGCCAATAGCCGCACAAGTCAGCTGTCCATATCAACACTGTTGGAACTGTGCAA AGGCCAAGCAGGAGAGTTGGCAGTTGGCAGAGAAATACTAAAAGCTGGATCCATTGGTATTGGTGGTGTTGATTATGTCTTA AATTGTATTCTTGGAAACCAAACTGAATCAAACAATTGGCAAGAACTTCTTGGCCGCCTTTGTCTTATAGATAGACTGTTGT TGGAATTTCCTGCTGAATTTTATCCTCATATTGTCAGTACTGATGTTTCACAAGCTGAGCCTGTTGAAATCAGGTATAAGAA GCTGCTGTCCCTCTTAACCTTTGCTTTGCAGTCCATTGATAATTCCCACTCAATGGTTGGCAAACTTTCCAGAAGGATCTAC TTGAGTTCTGCAAGAATGGTTACTACAGTACCCCATGTGTTTTCAAAACTGTTAGAAATGCTGAGTGTTTCCAGTTCCACTC ACTTCACCAGGATGCGTCGCCGTTTGATGGCTATTGCAGATGAGGTGGAAATTGCCGAAGCCATCCAGTTGGGCGTAGAAGA CACTTTGGATGGTCAACAGGACAGCTTCTTGCAGGCATCTGTTCCCAACAACTATCTGGAAACCACAGAGAACAGTTCCCCT GAGTGCACAGTCCATTTAGAGAAAACTGGAAAAGGATTATGTGCTACAAAATTGAGTGCCAGTTCAGAGGACATTTCTGAGA GACTGGCCAGCATTTCAGTAGGACCTTCTAGTTCAACAACAACAACAACAACAACAACAGAGCAACCAAAGCCAATGGTTCA AACAAAAGGCAGACCCCACAGTCAGTGTTTGAACTCCTCTCCTTTATCTCATCATTCCCAATTAATGTTTCCAGCCTTGTCA ACCCCTTCTTCTTCTACCCCATCTGTACCAGCTGGCACTGCAACAGATGTCTCTAAGCATAGACTTCAGGGATTCATTCCCT GCAGAATACCTTCTGCATCTCCTCAAACACAGCGCAAGTTTTCTCTACAATTCCACAGAAACTGTCCTGAAAACAAAGACTC AGATAAACTTTCCCCAGTCTTTACTCAGTCAAGACCCTTGCCCTCCAGTAACATACACAGGCCAAAGCCATCTAGACCTACC CCAGGTAATACAAGTAAACAGGGAGATCCCTCAAAAAATAGCATGACACTTGATCTGAACAGTAGTTCCAAATGTGATGACA GCTTTGGCTGTAGCAGCAATAGTAGTAATGCTGTTATACCCAGTGACGAGACAGTGTTCACCCCAGTAGAGGAGAAATGCAG ATTAGATGTCAATACAGAGCTCAACTCCAGTATTGAGGACCTTCTTGAAGCATCTATGCCTTCAAGTGATACAACAGTAACT TTTAAGTCAGAAGTTGCTGTCCTGTCTCCTGAAAAGGCTGAAAATGATGATACCTACAAAGATGATGTGAATCATAATCAAA AGTGCAAAGAGAAGATGGAAGCTGAAGAAGAAGAAGCTTTAGCAATTGCCATGGCAATGTCAGCGTCTCAGGATGCCCTCCC CATAGTTCCTCAGCTGCAGGTTGAAAATGGAGAAGATATCATCATTATTCAACAGGATACACCAGAGACTCTACCAGGACAT ACCAAAGCAAAACAACCGTATAGAGAAGACACTGAATGGCTGAAAGGTCAACAGATAGGCCTTGGAGCATTTTCTTCTTGTT ATCAGGCTCAAGATGTGGGAACTGGAACTTTAATGGCTGTTAAACAGGTGACTTATGTCAGAAACACATCTTCTGAGCAAGA AGAAGTAGTAGAAGCACTAAGAGAAGAGATAAGAATGATGAGCCATCTGAATCATCCAAACATCATTAGGATGTTGGGAGCC ACGTGTGAGAAGAGCAATTACAATCTCTTCATTGAATGGATGGCAGGGGGATCGGTGGCTCATTTGCTGAGTAAATATGGAG CCTTCAAAGAATCAGTAGTTATTAACTACACTGAACAGTTACTCCGTGGCCTTTCGTATCTCCATGAAAACCAAATCATTCA CAGAGATGTCAAAGGTGCCAATTTGCTAATTGACAGCACTGGTCAGAGACTAAGAATTGCAGATTTTGGAGCTGCAGCCAGG TTGGCATCAAAAGGAACTGGTGCAGGAGAGTTTCAGGGACAATTACTGGGGACAATTGCATTTATGGCACCTGAGGTACTAA GAGGTCAACAGTATGGAAGGAGCTGTGATGTATGGAGTGTTGGCTGTGCTATTATAGAAATGGCTTGTGCAAAACCACCATG GAATGCAGAAAAACACTCCAATCATCTTGCTTTGATATTTAAGATTGCTAGTGCAACTACTGCTCCATCGATCCCTTCACAT TTGTCTCCTGGTTTACGAGATGTGGCTCTTCGTTGTTTAGAACTTCAACCTCAGGACAGACCTCCATCAAGAGAGCTACTGA AGCATCCAGTCTTTCGTACTACATGGTAG HumanMEKK4CDS (SEQIDNO:19) ATGAGAGAAGCCGCTGCCGCGCTGGTCCCTCCTCCCGCCTTTGCCGTCACGCCTGCCGCCGCCATGGAGGAGCCGCCGCCAC CGCCGCCGCCGCCACCACCGCCACCGGAACCCGAGACCGAGTCAGAACCCGAGTGCTGCTTGGCGGCGAGGCAAGAGGGCAC ATTGGGAGATTCAGCTTGCAAGAGTCCTGAATCTGATCTAGAAGACTTCTCCGATGAAACAAATACAGAGAATCTTTATGGT ACCTCTCCCCCCAGCACACCTCGACAGATGAAACGCATGTCAACCAAACATCAGAGGAATAATGTGGGGAGGCCAGCCAGTC GGTCTAATTTGAAAGAAAAAATGAATGCACCAAATCAGCCTCCACATAAAGACACTGGAAAAACAGTGGAGAATGTGGAAGA ATACAGCTATAAGCAGGAGAAAAAGATCCGAGCAGCTCTTAGAACAACAGAGCGTGATCATAAAAAAAATGTACAGTGCTCA TTCATGTTAGACTCAGTGGGTGGATCTTTGCCAAAAAAATCAATTCCAGATGTGGATCTCAATAAGCCTTACCTCAGCCTTG GCTGTAGCAATGCTAAGCTTCCAGTATCTGTGCCCATGCCTATAGCCAGACCTGCACGCCAGACTTCTAGGACTGACTGTCC AGCAGATCGTTTAAAGTTTTTTGAAACTTTACGACTTTTGCTAAAGCTTACCTCAGTCTCAAAGAAAAAAGACAGGGAGCAA AGAGGACAAGAAAATACGTCTGGTTTCTGGCTTAACCGATCTAACGAACTGATCTGGTTAGAGCTACAAGCCTGGCATGCAG GACGGACAATTAACGACCAGGACTTCTTTTTATATACAGCCCGTCAAGCCATCCCAGATATTATTAATGAAATCCTTACTTT CAAAGTCGACTATGGGAGCTTCGCCTTTGTTAGAGATAGAGCTGGTTTTAATGGTACTTCAGTAGAAGGGCAGTGCAAAGCC ACTCCTGGAACAAAGATTGTAGGTTACTCAACACATCATGAGCATCTCCAACGCCAGAGGGTCTCATTTGAGCAGGTAAAAC GGATAATGGAGCTGCTAGAGTACATAGAAGCACTTTATCCATCATTGCAGGCTCTTCAGAAGGACTATGAAAAATATGCTGC AAAAGACTTCCAGGACAGGGTGCAGGCACTCTGTTTGTGGTTAAACATCACAAAAGACTTAAATCAGAAATTAAGGATTATG GGCACTGTTTTGGGCATCAAGAATTTATCAGACATTGGCTGGCCAGTGTTTGAAATCCCTTCCCCTCGACCATCCAAAGGTA ATGAGCCGGAGTATGAGGGTGATGACACAGAAGGAGAATTAAAGGAGTTGGAAAGTAGTACGGATGAGAGTGAAGAAGAACA AATCTCTGATCCTAGGGTACCGGAAATCAGACAGCCCATAGATAACAGCTTCGACATCCAGTCGCGGGACTGCATATCCAAG AAGCTTGAGAGGCTCGAATCTGAGGATGATTCTCTTGGCTGGGGAGCACCAGACTGGAGCACAGAAGCAGGCTTTAGTAGAC ATTGTCTGACTTCTATTTATAGACCATTTGTAGACAAAGCACTGAAGCAGATGGGGTTAAGAAAGTTAATTTTAAGACTTCA CAAGCTAATGGATGGTTCCTTGCAAAGGGCACGTATAGCATTGGTAAAGAACGATCGTCCAGTGGAGTTTTCTGAATTTCCA GATCCCATGTGGGGTTCAGATTATGTGCAGTTGTCAAGGACACCACCTTCATCTGAGGAGAAATGCAGTGCTGTGTCGTGGG AGGAGCTGAAGGCCATGGATTTACCTTCATTCGAACCTGCCTTCCTAGTTCTCTGCCGAGTCCTTCTGAATGTCATACATGA GTGTCTGAAGTTAAGATTGGAGCAGAGACCTGCTGGAGAACCATCTCTCTTGAGTATTAAGCAGCTGGTGAGAGAGTGTAAG GAGGTCCTGAAGGGCGGCCTGCTGATGAAGCAGTACTACCAGTTCATGCTGCAGGAGGTTCTGGAGGACTTGGAGAAGCCCG ACTGCAACATTGACGCTTTTGAAGAGGATCTACATAAAATGCTTATGGTGTATTTTGATTACATGAGAAGCTGGATCCAAAT GCTACAGCAATTACCTCAAGCATCGCATAGTTTAAAAAATCTGTTAGAAGAAGAATGGAATTTCACCAAAGAAATAACTCAT TACATACGGGGAGGAGAAGCACAGGCCGGGAAGCTTTTCTGTGACATTGCAGGAATGCTGCTGAAATCTACAGGAAGTTTTT TAGAATTTGGCTTACAGGAGAGCTGTGCTGAATTTTGGACTAGTGCGGATGACAGCAGTGCTTCCGACGAAATCAGGAGGTC TGTTATAGAGATCAGTCGAGCCCTGAAGGAGCTCTTCCATGAAGCCAGAGAAAGGGCTTCCAAAGCACTTGGATTTGCTAAA ATGTTGAGAAAGGACCTGGAAATAGCAGCAGAATTCAGGCTTTCAGCCCCAGTTAGAGACCTCCTGGATGTTCTGAAATCAA AACAGTATGTCAAGGTGCAAATTCCTGGGTTAGAAAACTTGCAAATGTTTGTTCCAGACACTCTTGCTGAGGAGAAGAGTAT TATTTTGCAGTTACTCAATGCAGCTGCAGGAAAGGACTGTTCAAAAGATTCAGATGACGTACTCATCGATGCCTATCTGCTT CTGACCAAGCACGGTGATCGAGCCCGTGATTCAGAGGACAGCTGGGGCACCTGGGAGGCACAGCCTGTCAAAGTCGTGCCTC AGGTGGAGACTGTTGACACCCTGAGAAGCATGCAGGTGGATAATCTTTTACTAGTTGTCATGCAGTCTGCGCATCTCACAAT TCAGAGAAAAGCTTTCCAGCAGTCCATTGAGGGACTTATGACTCTGTGCCAGGAGCAGACATCCAGTCAGCCGGTCATCGCC AAAGCTTTGCAGCAGCTGAAGAATGATGCATTGGAGCTATGCAACAGGATAAGCAATGCCATTGACCGCGTGGACCACATGT TCACATCAGAATTTGATGCTGAGGTTGATGAATCTGAATCTGTCACCTTGCAACAGTACTACCGAGAAGCAATGATTCAGGG GTACAATTTTGGATTTGAGTATCATAAAGAAGTTGTTCGTTTGATGTCTGGGGAGTTTAGACAGAAGATAGGAGACAAATAT ATAAGCTTTGCCCGGAAGTGGATGAATTATGTCCTGACTAAATGTGAGAGTGGTAGAGGTACAAGACCCAGGTGGGCGACTC AAGGATTTGATTTTCTACAAGCAATTGAACCTGCCTTTATTTCAGCTTTACCAGAAGATGACTTCTTGAGTTTACAAGCCTT GATGAATGAATGCATTGGCCATGTCATAGGAAAACCACACAGTCCTGTTACAGGTTTGTACCTTGCCATTCATCGGAACAGC CCCCGTCCTATGAAGGTACCTCGATGCCATAGTGACCCTCCTAACCCACACCTCATTATCCCCACTCCAGAGGGATTCAGCA CTCGGAGCATGCCTTCCGACGCGCGGAGCCATGGCAGCCCTGCTGCTGCTGCTGCTGCTGCTGCTGCTGCTGTTGCTGCCAG TCGGCCCAGCCCCTCTGGTGGTGACTCTGTGCTGCCCAAATCCATCAGCAGTGCCCATGATACCAGGGGTTCCAGCGTTCCT GAAAATGATCGATTGGCTTCCATAGCTGCTGAATTGCAGTTTAGGTCCCTGAGTCGTCACTCAAGCCCCACGGAGGAGCGAG ATGAACCAGCATATCCAAGAGGAGATTCAAGTGGGTCCACAAGAAGAAGTTGGGAACTTCGGACACTAATCAGCCAGAGTAA AGATACTGCTTCTAAACTAGGACCCATAGAAGCTATCCAGAAGTCAGTCCGATTGTTTGAAGAAAAGAGGTACCGAGAAATG AGGAGAAAGAATATCATTGGTCAAGTTTGTGATACGCCTAAGTCCTATGATAATGTTATGCACGTTGGCTTGAGGAAGGTGA CCTTCAAATGGCAAAGAGGAAACAAAATTGGAGAAGGCCAGTATGGGAAGGTGTACACCTGCATCAGCGTCGACACCGGGGA GCTGATGGCCATGAAAGAGATTCGATTTCAACCTAATGACCATAAGACTATCAAGGAAACTGCAGACGAATTGAAAATATTC GAAGGCATCAAACACCCCAATCTGGTTCGGTATTTTGGTGTGGAGCTCCATAGAGAAGAAATGTACATCTTCATGGAGTACT GCGATGAGGGGACTTTAGAAGAGGTGTCAAGGCTGGGACTTCAGGAACATGTGATTAGGCTGTATTCAAAGCAGATCACCAT TGCGATCAACGTCCTCCATGAGCATGGCATAGTCCACCGTGACATTAAAGGTGCCAATATCTTCCTTACCTCATCTGGATTA ATCAAACTGGGAGATTTTGGATGTTCAGTAAAGCTCAAAAACAATGCCCAGACCATGCCTGGTGAAGTGAACAGCACCCTGG GGACAGCAGCATACATGGCACCTGAAGTCATCACTCGTGCCAAAGGAGAGGGCCATGGGCGTGCGGCCGACATCTGGAGTCT GGGGTGTGTTGTCATAGAGATGGTGACTGGCAAGAGGCCTTGGCATGAGTATGAGCACAACTTTCAAATTATGTATAAAGTG GGGATGGGACATAAGCCACCAATCCCTGAAAGATTAAGCCCTGAAGGAAAGGACTTCCTTTCTCACTGCCTTGAGAGTGACC CAAAGATGAGATGGACCGCCAGCCAGCTCCTCGACCATTCGTTTGTCAAGGTTTGCACAGATGAAGAATGA HumanMEKK7CDS (SEQIDNO:20) ATGTCTACAGCCTCTGCCGCCTCCTCCTCCTCCTCGTCTTCGGCCGGTGAGATGATCGAAGCCCCTTCCCAGGTCCTCAACT TTGAAGAGATCGACTACAAGGAGATCGAGGTGGAAGAGGTTGTTGGAAGAGGAGCCTTTGGAGTTGTTTGCAAAGCTAAGTG GAGAGCAAAAGATGTTGCTATTAAACAAATAGAAAGTGAATCTGAGAGGAAAGCGTTTATTGTAGAGCTTCGGCAGTTATCC CGTGTGAACCATCCTAATATTGTAAAGCTTTATGGAGCCTGCTTGAATCCAGTGTGTCTTGTGATGGAATATGCTGAAGGGG GCTCTTTATATAATGTGCTGCATGGTGCTGAACCATTGCCATATTATACTGCTGCCCACGCAATGAGTTGGTGTTTACAGTG TTCCCAAGGAGTGGCTTATCTTCACAGCATGCAACCCAAAGCGCTAATTCACAGGGACCTGAAACCACCAAACTTACTGCTG GTTGCAGGGGGGACAGTTCTAAAAATTTGTGATTTTGGTACAGCCTGTGACATTCAGACACACATGACCAATAACAAGGGGA GTGCTGCTTGGATGGCACCTGAAGTTTTTGAAGGTAGTAATTACAGTGAAAAATGTGACGTCTTCAGCTGGGGTATTATTCT TTGGGAAGTGATAACGCGTCGGAAACCCTTTGATGAGATTGGTGGCCCAGCTTTCCGAATCATGTGGGCTGTTCATAATGGT ACTCGACCACCACTGATAAAAAATTTACCTAAGCCCATTGAGAGCCTGATGACTCGTTGTTGGTCTAAAGATCCTTCCCAGC GCCCTTCAATGGAGGAAATTGTGAAAATAATGACTCACTTGATGCGGTACTTTCCAGGAGCAGATGAGCCATTACAGTATCC TTGTCAGTATTCAGATGAAGGACAGAGCAACTCTGCCACCAGTACAGGCTCATTCATGGACATTGCTTCTACAAATACGAGT AACAAAAGTGACACTAATATGGAGCAAGTTCCTGCCACAAATGATACTATTAAGCGCTTAGAATCAAAATTGTTGAAAAATC AGGCAAAGCAACAGAGTGAATCTGGACGTTTAAGCTTGGGAGCCTCCCGTGGGAGCAGTGTGGAGAGCTTGCCCCCAACCTC TGAGGGCAAGAGGATGAGTGCTGACATGTCTGAAATAGAAGCTAGGATCGCCGCAACCACAGGCAACGGACAGCCAAGACGT AGATCCATCCAAGACTTGACTGTAACTGGAACAGAACCTGGTCAGGTGAGCAGTAGGTCATCCAGTCCCAGTGTCAGAATGA TTACTACCTCAGGACCAACCTCAGAAAAGCCAACTCGAAGTCATCCATGGACCCCTGATGATTCCACAGATACCAATGGATC AGATAACTCCATCCCAATGGCTTATCTTACACTGGATCACCAACTACAGCCTCTAGCACCGTGCCCAAACTCCAAAGAATCT ATGGCAGTGTTTGAACAGCATTGTAAAATGGCACAAGAATATATGAAAGTTCAAACAGAAATTGCATTGTTATTACAGAGAA AGCAAGAACTAGTTGCAGAACTGGACCAGGATGAAAAGGACCAGCAAAATACATCTCGCCTGGTACAGGAACATAAAAAGCT TTTAGATGAAAACAAAAGCCTTTCTACTTACTACCAGCAATGCAAAAAACAACTAGAGGTCATCAGAAGTCAGCAGCAGAAA CGACAAGGCACTTCATGA HumanJNKCDS (SEQIDNO:21) ATGAGCAGAAGCAAGCGTGACAACAATTTTTATAGTGTAGAGATTGGAGATTCTACATTCACAGTCCTGAAACGATATCAGA ATTTAAAACCTATAGGCTCAGGAGCTCAAGGAATAGTATGCGCAGCTTATGATGCCATTCTTGAAAGAAATGTTGCAATCAA GAAGCTAAGCCGACCATTTCAGAATCAGACTCATGCCAAGCGGGCCTACAGAGAGCTAGTTCTTATGAAATGTGTTAATCAC AAAAATATAATTGGCCTTTTGAATGTTTTCACACCACAGAAATCCCTAGAAGAATTTCAAGATGTTTACATAGTCATGGAGC TCATGGATGCAAATCTTTGCCAAGTGATTCAGATGGAGCTAGATCATGAAAGAATGTCCTACCTTCTCTATCAGATGCTGTG TGGAATCAAGCACCTTCATTCTGCTGGAATTATTCATCGGGACTTAAAGCCCAGTAATATAGTAGTAAAATCTGATTGCACT TTGAAGATTCTTGACTTCGGTCTGGCCAGGACTGCAGGAACGAGTTTTATGATGACGCCTTATGTAGTGACTCGCTACTACA GAGCACCCGAGGTCATCCTTGGCATGGGCTACAAGGAAAACGTTGACATTTGGTCAGTTGGGTGCATCATGGGAGAAATGAT CAAAGGTGGTGTTTTGTTCCCAGGTACAGATCATATTGATCAGTGGAATAAAGTTATTGAACAGCTTGGAACACCATGTCCT GAATTCATGAAGAAACTGCAACCAACAGTAAGGACTTACGTTGAAAACAGACCTAAATATGCTGGATATAGCTTTGAGAAAC TCTTCCCTGATGTCCTTTTCCCAGCTGACTCAGAACACAACAAACTTAAAGCCAGTCAGGCAAGGGATTTGTTATCCAAAAT GCTGGTAATAGATGCATCTAAAAGGATCTCTGTAGATGAAGCTCTCCAACACCCGTACATCAATGTCTGGTATGATCCTTCT GAAGCAGAAGCTCCACCACCAAAGATCCCTGACAAGCAGTTAGATGAAAGGGAACACACAATAGAAGAGTGGAAAGAATTGA TATATAAGGAAGTTATGGACTTGGAGGAGAGAACCAAGAATGGAGTTATACGGGGGCAGCCCTCTCCTTTAGGTGCAGCAGT GATCAATGGCTCTCAGCATCCATCATCATCGTCGTCTGTCAATGATGTGTCTTCAATGTCAACAGATCCGACTTTGGCCTCT GATACAGACAGCAGTCTAGAAGCAGCAGCTGGGCCTCTGGGCTGCTGTAGATGA HumanAP-1CDS (SEQIDNO:22) ATGACTGCAAAGATGGAAACGACCTTCTATGACGATGCCCTCAACGCCTCGTTCCTCCCGTCCGAGAGCGGACCTTATGGCT ACAGTAACCCCAAGATCCTGAAACAGAGCATGACCCTGAACCTGGCCGACCCAGTGGGGAGCCTGAAGCCGCACCTCCGCGC CAAGAACTCGGACCTCCTCACCTCGCCCGACGTGGGGCTGCTCAAGCTGGCGTCGCCCGAGCTGGAGCGCCTGATAATCCAG TCCAGCAACGGGCACATCACCACCACGCCGACCCCCACCCAGTTCCTGTGCCCCAAGAACGTGACAGATGAGCAGGAGGGCT TCGCCGAGGGCTTCGTGCGCGCCCTGGCCGAACTGCACAGCCAGAACACGCTGCCCAGCGTCACGTCGGCGGCGCAGCCGGT CAACGGGGCAGGCATGGTGGCTCCCGCGGTAGCCTCGGTGGCAGGGGGCAGCGGCAGCGGCGGCTTCAGCGCCAGCCTGCAC AGCGAGCCGCCGGTCTACGCAAACCTCAGCAACTTCAACCCAGGCGCGCTGAGCAGCGGCGGCGGGGCGCCCTCCTACGGCG CGGCCGGCCTGGCCTTTCCCGCGCAACCCCAGCAGCAGCAGCAGCCGCCGCACCACCTGCCCCAGCAGATGCCCGTGCAGCA CCCGCGGCTGCAGGCCCTGAAGGAGGAGCCTCAGACAGTGCCCGAGATGCCCGGCGAGACACCGCCCCTGTCCCCCATCGAC ATGGAGTCCCAGGAGCGGATCAAGGCGGAGAGGAAGCGCATGAGGAACCGCATCGCTGCCTCCAAGTGCCGAAAAAGGAAGC TGGAGAGAATCGCCCGGCTGGAGGAAAAAGTGAAAACCTTGAAAGCTCAGAACTCGGAGCTGGCGTCCACGGCCAACATGCT CAGGGAACAGGTGGCACAGCTTAAACAGAAAGTCATGAACCACGTTAACAGTGGGTGCCAACTCATGCTAACGCAGCAGTTG CAAACATTTTGA HumanASK1CDS (SEQIDNO:23) ATGAGCACGGAGGCGGACGAGGGCATCACTTTCTCTGTGCCACCCTTCGCCCCCTCGGGCTTCTGCACCATCCCCGAGGGCG GCATCTGCAGGAGGGGAGGAGCGGCGGCGGTGGGCGAGGGCGAGGAGCACCAGCTGCCACCGCCGCCGCCGGGCAGCTTCTG GAACGTGGAGAGCGCCGCTGCCCCTGGCATCGGTTGTCCGGCGGCCACCTCCTCGAGCAGTGCCACCCGAGGCCGGGGCAGC TCTGTTGGCGGGGGCAGCCGACGGACCACGGTGGCATATGTGATCAACGAAGCGAGCCAAGGGCAACTGGTGGTGGCCGAGA GCGAGGCCCTGCAGAGCTTGCGGGAGGCGTGCGAGACAGTGGGCGCCACCCTGGAAACCCTGCATTTTGGGAAACTCGACTT TGGAGAAACCACCGTGCTGGACCGCTTTTACAATGCAGATATTGCGGTGGTGGAGATGAGCGATGCCTTCCGGCAGCCGTCC TTGTTTTACCACCTTGGGGTGAGAGAAAGTTTCAGCATGGCCAACAACATCATCCTCTACTGTGATACTAACTCGGACTCTC TGCAGTCACTGAAGGAAATAATTTGCCAGAAGAATACTATGTGCACTGGGAACTACACCTTTGTTCCTTACATGATAACTCC ACATAACAAAGTCTACTGCTGTGACAGCAGCTTCATGAAGGGGTTGACAGAGCTCATGCAACCGAACTTCGAGCTGCTTCTT GGACCCATCTGCTTACCTCTTGTGGATCGTTTTATTCAACTTTTGAAGGTGGCACAAGCAAGTTCTAGCCAGTACTTCCGGG AATCTATACTCAATGACATCAGGAAAGCTCGTAATTTATACACTGGTAAAGAATTGGCAGCTGAGTTGGCAAGAATTCGGCA GCGAGTAGATAATATCGAAGTCTTGACAGCAGATATTGTCATAAATCTGTTACTTTCCTACAGAGATATCCAGGACTATGAT TCTATTGTGAAGCTGGTAGAGACTTTAGAAAAACTGCCAACCTTTGATTTGGCCTCCCATCACCATGTGAAGTTTCATTATG CATTTGCACTGAATAGGAGAAATCTCCCTGGTGACAGAGCAAAAGCTCTTGATATTATGATTCCCATGGTGCAAAGCGAAGG ACAAGTTGCTTCAGATATGTATTGCCTAGTTGGTCGAATCTACAAAGATATGTTTTTGGACTCTAATTTCACGGACACTGAA AGCAGAGACCATGGAGCTTCTTGGTTCAAAAAGGCATTTGAATCTGAGCCAACACTACAGTCAGGAATTAATTATGCGGTCC TCCTCCTGGCAGCTGGACACCAGTTTGAATCTTCCTTTGAGCTCCGGAAAGTTGGGGTGAAGCTAAGTAGTCTTCTTGGTAA AAAGGGAAACTTGGAAAAACTCCAGAGCTACTGGGAAGTTGGATTTTTTCTGGGGGCCAGCGTCCTAGCCAATGACCACATG AGAGTCATTCAAGCATCTGAAAAGCTTTTTAAACTGAAGACACCAGCATGGTACCTCAAGTCTATTGTAGAGACAATTTTAA TATATAAGCATTTTGTGAAACTGACCACAGAACAGCCTGTGGCCAAGCAAGAACTTGTGGACTTTTGGATGGATTTCCTGGT CGAGGCCACAAAGACAGATGTTACTGTGGTTAGGTTTCCAGTATTAATATTAGAACCAACCAAAATCTATCAACCTTCTTAT TTGTCTATCAACAATGAAGTTGAGGAAAAGACAATCTCTATTTGGCACGTGCTTCCTGATGACAAGAAAGGTATACATGAGT GGAATTTTAGTGCCTCTTCTGTCAGGGGAGTGAGTATTTCTAAATTTGAAGAAAGATGCTGCTTTCTTTATGTGCTTCACAA TTCTGATGATTTCCAAATCTATTTCTGTACAGAACTTCATTGTAAAAAGTTTTTTGAGATGGTGAACACCATTACCGAAGAG AAGGGGAGAAGCACAGAGGAAGGAGACTGTGAAAGTGACTTGCTGGAGTATGACTATGAATATGATGAAAATGGTGACAGAG TCGTTTTAGGAAAAGGCACTTATGGGATAGTCTACGCAGGTCGGGACTTGAGCAACCAAGTCAGAATTGCTATTAAGGAAAT CCCAGAGAGAGACAGCAGATACTCTCAGCCCCTGCATGAAGAAATAGCATTGCATAAACACCTGAAGCACAAAAATATTGTC CAGTATCTGGGCTCTTTCAGTGAGAATGGTTTCATTAAAATCTTCATGGAGCAGGTCCCTGGAGGAAGTCTTTCTGCTCTCC TTCGTTCCAAATGGGGTCCATTAAAAGACAATGAGCAAACAATTGGCTTTTATACAAAGCAAATACTGGAAGGATTAAAATA TCTCCATGACAATCAGATAGTTCACCGGGACATAAAGGGTGACAATGTGTTGATTAATACCTACAGTGGTGTTCTCAAGATC TCTGACTTCGGAACATCAAAGAGGCTTGCTGGCATAAACCCCTGTACTGAAACTTTTACTGGTACCCTCCAGTATATGGCAC CAGAAATAATAGATAAAGGACCAAGAGGCTACGGAAAAGCAGCAGACATCTGGTCTCTGGGCTGTACAATCATTGAAATGGC CACAGGAAAACCCCCATTTTATGAACTGGGAGAACCACAAGCAGCTATGTTCAAGGTGGGAATGTTTAAAGTCCACCCTGAG ATCCCAGAGTCCATGTCTGCAGAGGCCAAGGCATTCATACTGAAATGTTTTGAACCAGATCCTGACAAGAGAGCCTGTGCTA ACGACTTGCTTGTTGATGAGTTTTTAAAAGTTTCAAGCAAAAAGAAAAAGACACAACCTAAGCTTTCAGCTCTTTCAGCTGG ATCAAATGAATATCTCAGGAGTATATCCTTGCCGGTACCTGTGCTGGTGGAGGACACCAGCAGCAGCAGTGAGTACGGCTCA GTTTCACCCGACACGGAGTTGAAAGTGGACCCCTTCTCTTTCAAAACAAGAGCCAAGTCCTGCGGAGAAAGAGATGTCAAGG GAATTCGGACACTCTTTTTGGGCATTCCAGATGAGAATTTTGAAGATCACAGTGCTCCTCCTTCCCCTGAAGAAAAAGATTC TGGATTCTTCATGCTGAGGAAGGACAGTGAGAGGCGAGCTACCCTTCACAGGATCCTGACGGAAGACCAAGACAAAATTGTG AGAAACCTAATGGAATCTTTAGCTCAGGGGGCTGAAGAACCGAAACTAAAATGGGAACACATCACAACCCTCATTGCAAGCC TCAGAGAATTTGTGAGATCCACTGACCGAAAAATCATAGCCACCACACTGTCAAAGCTGAAACTGGAGCTGGACTTCGACAG CCATGGCATTAGCCAAGTCCAGGTGGTACTCTTTGGTTTTCAAGATGCTGTCAATAAAGTTCTTCGGAATCATAACATCAAG CCGCACTGGATGTTTGCCTTAGACAGTATCATTCGGAAGGCGGTACAGACAGCCATTACCATCCTGGTTCCAGAACTAAGGC CACATTTCAGCCTTGCATCTGAGAGTGATACTGCTGATCAAGAAGACTTGGATGTAGAAGATGACCATGAGGAACAGCCTTC AAATCAAACTGTCCGAAGACCTCAGGCTGTCATTGAAGATGCTGTGGCTACCTCAGGCGTGAGCACGCTCAGTTCTACTGTG TCTCATGATTCCCAGAGTGCTCACCGGTCACTGAATGTACAGCTTGGAAGGATGAAAATAGAAACCAATAGATTACTGGAAG AATTGGTTCGGAAAGAGAAAGAATTACAAGCACTCCTTCATCGAGCTATTGAAGAAAAAGACCAAGAAATTAAACACCTGAA GCTTAAGTCCCAACCCATAGAAATTCCTGAATTGCCTGTATTTCATCTAAATTCTTCTGGCACAAATACTGAAGATTCTGAA CTTACCGACTGGCTGAGAGTGAATGGAGCTGATGAAGACACTATAAGCCGGTTTTTGGCTGAAGATTATACACTATTGGATG TTCTCTACTATGTTACACGTGATGACTTAAAATGCTTGAGACTAAGGGGAGGGATGCTGTGCACACTGTGGAAGGCTATCAT TGACTTTCGAAACAAACAGACTTGA HumanRIPCDS (SEQIDNO:24) ATGTGGAGCAAACTGAATAATGAAGAGCACAATGAGCTGAGGGAAGTGGACGGCACCGCTAAGAAGAATGGCGGCACCCTCT ACTACATGGCGCCCGAGCACCTGAATGACGTCAACGCAAAGCCCACAGAGAAGTCGGATGTGTACAGCTTTGCTGTAGTACT CTGGGCGATATTTGCAAATAAGGAGCCATATGAAAATGCTATCTGTGAGCAGCAGTTGATAATGTGCATAAAATCTGGGAAC AGGCCAGATGTGGATGACATCACTGAGTACTGCCCAAGAGAAATTATCAGTCTCATGAAGCTCTGCTGGGAAGCGAATCCGG AAGCTCGGCCGACATTTCCTGGCATTGAAGAAAAATTTAGGCCTTTTTATTTAAGTCAATTAGAAGAAAGTGTAGAAGAGGA CGTGAAGAGTTTAAAGAAAGAGTATTCAAACGAAAATGCAGTTGTGAAGAGAATGCAGTCTCTTCAACTTGATTGTGTGGCA GTACCTTCAAGCCGGTCAAATTCAGCCACAGAACAGCCTGGTTCACTGCACAGTTCCCAGGGACTTGGGATGGGTCCTGTGG AGGAGTCCTGGTTTGCTCCTTCCCTGGAGCACCCACAAGAAGAGAATGAGCCCAGCCTGCAGAGTAAACTCCAAGACGAAGC CAACTACCATCTTTATGGCAGCCGCATGGACAGGCAGACGAAACAGCAGCCCAGACAGAATGTGGCTTACAACAGAGAGGAG GAAAGGAGACGCAGGGTCTCCCATGACCCTTTTGCACAGCAAAGACCTTACGAGAATTTTCAGAATACAGAGGGAAAAGGCA CTGCTTATTCCAGTGCAGCCAGTCATGGTAATGCAGTGCACCAGCCCTCAGGGCTCACCAGCCAACCTCAAGTACTGTATCA GAACAATGGATTATATAGCTCACATGGCTTTGGAACAAGACCACTGGATCCAGGAACAGCAGGTCCCAGAGTTTGGTACAGG CCAATTCCAAGTCATATGCCTAGTCTGCATAATATCCCAGTGCCTGAGACCAACTATCTAGGAAATACACCCACCATGCCAT TCAGCTCCTTGCCACCAACAGATGAATCTATAAAATATACCATATACAATAGTACTGGCATTCAGATTGGAGCCTACAATTA TATGGAGATTGGTGGGACGAGTTCATCACTACTAGACAGCACAAATACGAACTTCAAAGAAGAGCCAGCTGCTAAGTACCAA GCTATCTTTGATAATACCACTAGTCTGACGGATAAACACCTGGACCCAATCAGGGAAAATCTGGGAAAGCACTGGAAAAACT GTGCCCGTAAACTGGGCTTCACACAGTCTCAGATTGATGAAATTGACCATGACTATGAGCGAGATGGACTGAAAGAAAAGGT TTACCAGATGCTCCAAAAGTGGGTGATGAGGGAAGGCATAAAGGGAGCCACGGTGGGGAAGCTGGCCCAGGCGCTCCACCAG TGTTCCAGGATCGACCTTCTGAGCAGCTTGATTTACGTCAGCCAGAACTAA HumanMEKK3CDS (SEQIDNO:25) ATGGACGAACAGGAGGCATTGAACTCAATCATGAACGATCTGGTGGCCCTCCAGATGAACCGACGTCACCGGATGCCTGGAT ATGAGACCATGAAGAACAAAGACACAGGTCACTCAAATAGGCAGAAAAAACACAACAGCAGCAGCTCAGCCCTTCTGAACAG CCCCACAGTAACAACAAGCTCATGTGCAGGGGCCAGTGAGAAAAAGAAATTTTTGAGTGACGTCAGAATCAAGTTCGAGCAC AACGGGGAGAGGCGAATTATAGCGTTCAGCCGGCCTGTGAAATATGAAGATGTGGAGCACAAGGTGACAACAGTATTTGGAC AACCTCTTGATCTACATTACATGAACAATGAGCTCTCCATCCTGCTGAAAAACCAAGATGATCTTGATAAAGCAATTGACAT TTTAGATAGAAGCTCAAGCATGAAAAGCCTTAGGATATTGCTGTTGTCCCAGGACAGAAACCATAACAGTTCCTCTCCCCAC TCTGGGGTGTCCAGACAGGTGCGGATCAAGGCTTCCCAGTCCGCAGGGGATATAAATACTATCTACCAGCCCCCCGAGCCCA GAAGCAGGCACCTCTCTGTCAGCTCCCAGAACCCTGGCCGAAGCTCACCTCCCCCTGGCTATGTTCCTGAGCGGCAGCAGCA CATTGCCCGGCAGGGGTCCTACACCAGCATCAACAGTGAGGGGGAGTTCATCCCAGAGACCAGCGAGCAGTGCATGCTGGAT CCCCTGAGCAGTGCAGAAAATTCCTTGTCTGGAAGCTGCCAATCCTTGGACAGGTCAGCAGACAGCCCATCCTTCCGGAAAT CACGAATGTCCCGTGCCCAGAGCTTCCCTGACAACAGACAGGAATACTCAGATCGGGAAACTCAGCTTTATGACAAAGGGGT CAAAGGTGGAACCTACCCCCGGCGCTACCACGTGTCTGTGCACCACAAGGACTACAGTGATGGCAGAAGAACATTTCCCCGA ATACGGCGTCATCAAGGCAACTTGTTCACCCTGGTGCCCTCCAGCCGCTCCCTGAGCACAAATGGCGAGAACATGGGTCTGG CTGTGCAATACCTGGACCCCCGTGGGCGCCTGCGGAGTGCGGACAGCGAGAATGCCCTCTCTGTGCAGGAGAGGAATGTGCC AACCAAGTCTCCCAGTGCCCCCATCAACTGGCGCCGGGGAAAGCTCCTGGGCCAGGGTGCCTTCGGCAGGGTCTATTTGTGC TATGACGTGGACACGGGACGTGAACTTGCTTCCAAGCAGGTCCAATTTGATCCAGACAGTCCTGAGACAAGCAAGGAGGTGA GTGCTCTGGAGTGCGAGATCCAGTTGCTAAAGAACTTGCAGCATGAGCGCATCGTGCAGTACTATGGCTGTCTGCGGGACCG CGCTGAGAAGACCCTGACCATCTTCATGGAGTACATGCCAGGGGGCTCGGTGAAAGACCAGTTGAAGGCTTACGGTGCTCTG ACAGAGAGCGTGACCCGAAAGTACACGCGGCAGATCCTGGAGGGCATGTCCTACCTGCACAGCAACATGATTGTTCACCGGG ACATTAAGGGAGCCAACATCCTCCGAGACTCTGCTGGGAATGTAAAGCTGGGGGACTTTGGGGCCAGCAAACGCCTGCAGAC GATCTGTATGTCGGGGACGGGCATGCGCTCCGTCACTGGCACACCCTACTGGATGAGCCCTGAGGTGATCAGCGGCGAGGGC TATGGAAGGAAAGCAGACGTGTGGAGCCTGGGCTGCACTGTGGTGGAGATGCTGACAGAGAAACCACCGTGGGCAGAGTATG AAGCTATGGCCGCCATCTTCAAGATTGCCACCCAGCCCACCAATCCTCAGCTGCCCTCCCACATCTCTGAACATGGCCGGGA CTTCCTGAGGCGCATTTTTGTGGAGGCTCGCCAGAGACCTTCAGCTGAGGAGCTGCTCACACACCACTTTGCACAGCTCATG TACTGA HumanMEKK6CDS (SEQIDNO:26) ATGGCGGGGCCGTGTCCCCGGTCCGGGGCGGAGCGCGCCGGCAGCTGCTGGCAGGACCCGCTGGCCGTGGCGCTGAGCCGGG GCCGGCAGCTCGCGGCGCCCCCGGGCCGGGGCTGCGCGCGGAGCCGGCCGCTCAGCGTGGTCTACGTGCTGACCCGGGAGCC GCAGCCCGGGCTCGAGCCTCGGGAGGGAACCGAGGCGGAGCCGCTGCCCCTGCGCTGCCTGCGCGAGGCTTGCGCGCAGGTC CCCCGGCCGCGGCCGCCCCCGCAGCTGCGCAGCCTGCCCTTCGGGACGCTGGAGCTAGGCGACACCGCGGCTCTGGATGCCT TCTACAACGCGGATGTGGTGGTGCTGGAGGTGAGCAGCTCGCTGGTACAGCCCTCCCTGTTCTACCACCTTGGTGTGCGTGA GAGCTTCAGCATGACCAACAATGTGCTCCTCTGCTCCCAGGCCGACCTCCCTGACCTGCAGGCCCTGCGGGAGGATGTTTTC CAGAAGAACTCGGATTGCGTTGGCAGCTACACACTGATCCCCTATGTGGTGACGGCCACTGGTCGGGTGCTGTGTGGTGATG CAGGCCTTCTGCGGGGCCTGGCTGATGGGCTGGTACAGGCTGGAGTGGGGACCGAGGCCCTGCTCACTCCCCTGGTGGGCCG GCTTGCCCGCCTGCTGGAGGCCACACCCACAGACTCTTGTGGCTATTTCCGGGAGACCATTCGGCGGGACATCCGGCAGGCG CGGGAGCGGTTCAGTGGGCCACAGCTGCGGCAGGAGCTGGCTCGCCTGCAGCGGAGACTGGACAGCGTGGAGCTGCTGAGCC CCGACATCATCATGAACTTGCTGCTCTCCTACCGCGATGTGCAGGACTACTCGGCCATCATTGAGCTGGTGGAGACGCTGCA GGCCTTGCCCACCTGTGATGTGGCCGAGCAGCATAATGTCTGCTTCCACTACACTTTTGCCCTCAACCGGAGGAACAGGCCT GGGGACCGGGCGAAGGCCCTGTCTGTGCTGCTGCCGCTGGTACAGCTTGAGGGCTCTGTGGCGCCCGATCTGTACTGCATGT GTGGCCGTATCTACAAGGACATGTTCTTCAGCTCGGGTTTCCAGGATGCTGGGCACCGGGAGCAGGCCTATCACTGGTATCG CAAGGCTTTTGACGTAGAGCCCAGCCTTCACTCAGGCATCAATGCAGCTGTGCTCCTCATTGCTGCCGGGCAGCACTTTGAG GATTCCAAAGAGCTCCGGCTAATAGGCATGAAGCTGGGCTGCCTGCTGGCCCGCAAAGGCTGCGTGGAGAAGATGCAGTATT ACTGGGATGTGGGTTTCTACCTGGGAGCCCAGATCCTCGCCAATGACCCCACCCAGGTGGTGCTGGCTGCAGAGCAGCTGTA TAAGCTCAATGCCCCCATATGGTACCTGGTGTCCGTGATGGAGACCTTCCTGCTCTACCAGCACTTCAGGCCCACGCCAGAG CCCCCTGGAGGGCCACCACGCCGTGCCCACTTCTGGCTCCACTTCTTGCTACAGTCCTGCCAACCATTCAAGACAGCCTGTG CCCAGGGCGACCAGTGCTTGGTGCTGGTCCTGGAGATGAACAAGGTGCTGCTGCCTGCAAAGCTCGAGGTTCGGGGTACTGA CCCAGTAAGCACAGTGACCCTGAGCCTGCTGGAGCCTGAGACCCAGGACATTCCCTCCAGCTGGACCTTCCCAGTCGCCTCC ATATGCGGAGTCAGCGCCTCAAAGCGCGACGAGCGCTGCTGCTTCCTCTATGCACTCCCCCCGGCTCAGGACGTCCAGCTGT GCTTCCCCAGCGTAGGGCACTGCCAGTGGTTCTGCGGCCTGATCCAGGCCTGGGTGACGAACCCGGATTCCACGGCGCCCGC GGAGGAGGCGGAGGGCGCGGGGGAGATGTTGGAGTTTGATTATGAGTACACGGAGACGGGCGAGCGGCTGGTGCTGGGCAAG GGCACGTATGGGGTGGTGTACGCGGGCCGCGATCGCCACACGAGGGTGCGCATCGCCATCAAGGAGATCCCGGAGCGGGACA GCAGGTTCTCTCAGCCCCTGCATGAAGAGATCGCTCTTCACAGACGCCTGCGCCACAAGAACATAGTGCGCTATCTGGGCTC AGCTAGCCAGGGCGGCTACCTTAAGATCTTCATGGAGGAAGTGCCTGGAGGCAGCCTGTCCTCCTTGCTGCGGTCGGTGTGG GGACCCCTGAAGGACAACGAGAGCACCATCAGTTTCTACACCCGCCAGATCCTGCAGGGACTTGGCTACTTGCACGACAACC ACATCGTGCACAGGGACATAAAAGGGGACAATGTGCTGATCAACACCTTCAGTGGGCTGCTCAAGATTTCTGACTTCGGCAC CTCCAAGCGGCTGGCAGGCATCACACCTTGCACTGAGACCTTCACAGGAACTCTGCAGTATATGGCCCCAGAAATCATTGAC CAGGGCCCACGCGGGTATGGGAAAGCAGCTGACATCTGGTCACTGGGCTGCACTGTCATTGAGATGGCCACAGGTCGCCCCC CCTTCCACGAGCTCGGGAGCCCACAGGCTGCCATGTTTCAGGTGGGTATGTACAAGGTCCATCCGCCAATGCCCAGCTCTCT GTCGGCCGAGGCCCAAGCCTTTCTCCTCCGAACTTTTGAGCCAGACCCCCGCCTCCGAGCCAGCGCCCAGACACTGCTGGGG GACCCCTTCCTGCAGCCTGGGAAAAGGAGCCGCAGCCCCAGCTCCCCACGACATGCTCCACGGCCCTCAGATGCCCCTTCTG CCAGTCCCACTCCTTCAGCCAACTCAACCACCCAGTCTCAGACATTCCCGTGCCCTCAGGCACCCTCTCAGCACCCACCCAG CCCCCCGAAGCGCTGCCTCAGTTATGGGGGCACCAGCCAGCTCCGGGTGCCCGAGGAGCCTGCGGCCGAGGAGCCTGCGTCT CCGGAGGAGAGTTCGGGGCTGAGCCTGCTGCACCAGGAGAGCAAGCGTCGGGCCATGCTGGCCGCAGTATTGGAGCAGGAGC TGCCAGCGCTGGCGGAGAATCTGCACCAGGAGCAGAAGCAAGAGCAGGGGGCCCGTCTGGGCAGAAACCATGTGGAAGAGCT GCTGCGCTGCCTCGGGGCACACATCCACACTCCCAACCGCCGGCAGCTCGCCCAGGAGCTGCGGGCGCTGCAAGGACGGCTG AGGGCCCAGGGCCTTGGGCCTGCGCTTCTGCACAGACCGCTGTTTGCCTTCCCGGATGCGGTGAAGCAGATCCTCCGCAAGC GCCAGATCCGTCCACACTGGATGTTCGTTCTGGACTCACTGCTCAGCCGTGCTGTGCGGGCAGCCCTGGGTGTGCTAGGACC GGAGGTGGAGAAGGAGGCGGTCTCACCGAGGTCAGAGGAGCTGAGTAATGAAGGGGACTCCCAGCAGAGCCCAGGCCAGCAG AGCCCGCTTCCGGTGGAGCCCGAGCAGGGCCCCGCTCCTCTGATGGTGCAGCTGAGCCTCTTGAGGGCAGAGACTGATCGGC TGCGCGAAATCCTGGCGGGGAAGGAACGGGAGTACCAGGCCCTGGTGCAGCGGGCTCTACAGCGGCTGAATGAGGAAGCCCG GACCTATGTCCTGGCCCCAGAGCCTCCAACTGCTCTTTCAACGGACCAGGGCCTGGTGCAGTGGCTACAGGAACTGAATGTG GATTCAGGCACCATCCAAATGCTGTTGAACCATAGCTTCACCCTCCACACTCTGCTCACCTATGCCACTCGAGATGACCTCA TCTACACCCGCATCAGGGGAGGGATGGTATGCCGCATCTGGAGGGCCATCTTGGCACAGCGAGCAGGATCCACACCAGTCAC CTCTGGACCCTGA HumanNIKCDS (SEQIDNO:27) ATGGCAGTGATGGAAATGGCCTGCCCAGGTGCCCCTGGCTCAGCAGTGGGGCAGCAGAAGGAACTCCCCAAAGCCAAGGAGA AGACGCCGCCACTGGGGAAGAAACAGAGCTCCGTCTACAAGCTTGAGGCCGTGGAGAAGAGCCCTGTGTTCTGCGGAAAGTG GGAGATCCTGAATGACGTGATTACCAAGGGCACAGCCAAGGAAGGCTCCGAGGCAGGGCCAGCTGCCATCTCTATCATCGCC CAGGCTGAGTGTGAGAATAGCCAAGAGTTCAGCCCCACCTTTTCAGAACGCATTTTCATCGCTGGGTCCAAACAGTACAGCC AGTCCGAGAGTCTTGATCAGATCCCCAACAATGTGGCCCATGCTACAGAGGGCAAAATGGCCCGTGTGTGTTGGAAGGGAAA GCGTCGCAGCAAAGCCCGGAAGAAACGGAAGAAGAAGAGCTCAAAGTCCCTGGCTCATGCAGGAGTGGCCTTGGCCAAACCC CTCCCCAGGACCCCTGAGCAGGAGAGCTGCACCATCCCAGTGCAGGAGGATGAGTCTCCACTCGGCGCCCCATATGTTAGAA ACACCCCGCAGTTCACCAAGCCTCTGAAGGAACCAGGCCTTGGGCAACTCTGTTTTAAGCAGCTTGGCGAGGGCCTACGGCC GGCTCTGCCTCGATCAGAACTCCACAAACTGATCAGCCCCTTGCAATGTCTGAACCACGTGTGGAAACTGCACCACCCCCAG GACGGAGGCCCCCTGCCCCTGCCCACGCACCCCTTCCCCTATAGCAGACTGCCTCATCCCTTCCCATTCCACCCTCTCCAGC CCTGGAAACCTCACCCTCTGGAGTCCTTCCTGGGCAAACTGGCCTGTGTAGACAGCCAGAAACCCTTGCCTGACCCACACCT GAGCAAACTGGCCTGTGTAGACAGTCCAAAGCCCCTGCCTGGCCCACACCTGGAGCCCAGCTGCCTGTCTCGTGGTGCCCAT GAGAAGTTTTCTGTGGAGGAATACCTAGTGCATGCTCTGCAAGGCAGCGTGAGCTCAGGCCAGGCCCACAGCCTGACCAGCC TGGCCAAGACCTGGGCAGCAAGGGGCTCCAGATCCCGGGAGCCCAGCCCCAAAACTGAGGACAACGAGGGTGTCCTGCTCAC TGAGAAACTCAAGCCAGTGGATTATGAGTACCGAGAAGAAGTCCACTGGGCCACGCACCAGCTCCGCCTGGGCAGAGGCTCC TTCGGAGAGGTGCACAGGATGGAGGACAAGCAGACTGGCTTCCAGTGCGCTGTCAAAAAGGTGCGGCTGGAAGTATTTCGGG CAGAGGAGCTGATGGCATGTGCAGGATTGACCTCACCCAGAATTGTCCCTTTGTATGGAGCTGTGAGAGAAGGGCCTTGGGT CAACATCTTCATGGAGCTGCTGGAAGGTGGCTCCCTGGGCCAGCTGGTCAAGGAGCAGGGCTGTCTCCCAGAGGACCGGGCC CTGTACTACCTGGGCCAGGCCCTGGAGGGTCTGGAATACCTCCACTCACGAAGGATTCTGCATGGGGACGTCAAAGCTGACA ACGTGCTCCTGTCCAGCGATGGGAGCCACGCAGCCCTCTGTGACTTTGGCCATGCTGTGTGTCTTCAACCTGATGGCCTGGG AAAGTCCTTGCTCACAGGGGACTACATCCCTGGCACAGAGACCCACATGGCTCCGGAGGTGGTGCTGGGCAGGAGCTGCGAC GCCAAGGTGGATGTCTGGAGCAGCTGCTGTATGATGCTGCACATGCTCAACGGCTGCCACCCCTGGACTCAGTTCTTCCGAG GGCCGCTCTGCCTCAAGATTGCCAGCGAGCCTCCGCCTGTGAGGGAGATCCCACCCTCCTGCGCCCCTCTCACAGCCCAGGC CATCCAAGAGGGGCTGAGGAAAGAGCCCATCCACCGCGTGTCTGCAGCGGAGCTGGGAGGGAAGGTGAACCGGGCACTACAG CAAGTGGGAGGTCTGAAGAGCCCTTGGAGGGGAGAATATAAAGAACCAAGACATCCACCGCCAAATCAAGCCAATTACCACC AGACCCTCCATGCCCAGCCGAGAGAGCTTTCGCCAAGGGCCCCAGGGCCCCGGCCAGCTGAGGAGACAACAGGCAGAGCCCC TAAGCTCCAGCCTCCTCTCCCACCAGAGCCCCCAGAGCCAAACAAGTCTCCTCCCTTGACTTTGAGCAAGGAGGAGTCTGGG ATGTGGGAACCCTTACCTCTGTCCTCCCTGGAGCCAGCCCCTGCCAGAAACCCCAGCTCACCAGAGCGGAAAGCAACCGTCC CGGAGCAGGAACTGCAGCAGCTGGAAAJAGAATTATTCCTCAACAGCCTGTCCCAGCCATTTTCTCTGGAGGAGCAGGAGCA AATTCTCTCGTGCCTCAGCATCGACAGCCTCTCCCTGTCGGATGACAGTGAGAAGAACCCATCAAAGGCCTCTCAAAGCTCG CGGGACACCCTGAGCTCAGGCGTACACTCCTGGAGCAGCCAGGCCGAGGCTCGAAGCTCCAGCTGGAACATGGTGCTGGCCC GGGGGCGGCCCACCGACACCCCAAGCTATTTCAATGGTGTGAAAGTCCAAATACAGTCTCTTAATGGTGAACACCTGCACAT CCGGGAGTTCCACCGGGTCAAAGTGGGAGACATCGCCACTGGCATCAGCAGCCAGATCCCAGCTGCAGCCTTCAGCTTGGTC ACCAAAGACGGGCAGCCTGTTCGCTACGACATGGAGGTGCCAGACTCGGGCATCGACCTGCAGTGCACACTGGCCCCTGATG GCAGCTTCGCCTGGAGCTGGAGGGTCAAGCATGGCCAGCTGGAGAACAGGCCCTAA HumanIKKCDS (SEQIDNO:28) ATGTTTTCAGGGGGGTGTCATAGCCCCGGGTTTGGCCGCCCCAGCCCCGCCTTCCCCGCCCCGGGGAGCCCGCCCCCTGCCC CGCGTCCCTGCCGACAGGAAACAGGTGAGCAGATTGCCATCAAGCAGTGCCGGCAGGAGCTCAGCCCCCGGAACCGAGAGCG GTGGTGCCTGGAGATCCAGATCATGAGAAGGCTGACCCACCCCAATGTGGTGGCTGCCCGAGATGTCCCTGAGGGGATGCAG AACTTGGCGCCCAATGACCTGCCCCTGCTGGCCATGGAGTACTGCCAAGGAGGAGATCTCCGGAAGTACCTGAACCAGTTTG AGAACTGCTGTGGTCTGCGGGAAGGTGCCATCCTCACCTTGCTGAGTGACATTGCCTCTGCGCTTAGATACCTTCATGAAAA CAGAATCATCCATCGGGATCTAAAGCCAGAAAACATCGTCCTGCAGCAAGGAGAACAGAGGTTAATACACAAAATTATTGAC CTAGGATATGCCAAGGAGCTGGATCAGGGCAGTCTTTGCACATCATTCGTGGGGACCCTGCAGTACCTGGCCCCAGAGCTAC TGGAGCAGCAGAAGTACACAGTGACCGTCGACTACTGGAGCTTCGGCACCCTGGCCTTTGAGTGCATCACGGGCTTCCGGCC CTTCCTCCCCAACTGGCAGCCCGTGCAGTGGCATTCAAAAGTGCGGCAGAAGAGTGAGGTGGACATTGTTGTTAGCGAAGAC TTGAATGGAACGGTGAAGTTTTCAAGCTCTTTACCCTACCCCAATAATCTTAACAGTGTCCTGGCTGAGCGACTGGAGAAGT GGCTGCAACTGATGCTGATGTGGCACCCCCGACAGAGGGGCACGGATCCCACGTATGGGCCCAATGGCTGCTTCAAGGCCCT GGATGACATCTTAAACTTAAAGCTGGTTCATATCTTGAACATGGTCACGGGCACCATCCACACCTACCCTGTGACAGAGGAT GAGAGTCTGCAGAGCTTGAAGGCCAGAATCCAACAGGACACGGGCATCCCAGAGGAGGACCAGGAGCTGCTGCAGGAAGCGG GCCTGGCGTTGATCCCCGATAAGCCTGCCACTCAGTGTATTTCAGACGGCAAGTTAAATGAGGGCCACACATTGGACATGGA TCTTGTTTTTCTCTTTGACAACAGTAAAATCACCTATGAGACTCAGATCTCCCCACGGCCCCAACCTGAAAGTGTCAGCTGT ATCCTTCAAGAGCCCAAGAGGAATCTCGCCTTCTTCCAGCTGAGGAAGGTGTGGGGCCAGGTCTGGCACAGCATCCAGACCC TGAAGGAAGATTGCAACCGGCTGCAGCAGGGACAGCGAGCCGCCATGATGAATCTCCTCCGAAACAACAGCTGCCTCTCCAA AATGAAGAATTCCATGGCTTCCATGTCTCAGCAGCTCAAGGCCAAGTTGGATTTCTTCAAAACCAGCATCCAGATTGACCTG GAGAAGTACAGCGAGCAAACCGAGTTTGGGATCACATCAGATAAACTGCTGCTGGCCTGGAGGGAAATGGAGCAGGCTGTGG AGCTCTGTGGGCGGGAGAACGAAGTGAAACTCCTGGTAGAACGGATGATGGCTCTGCAGACCGACATTGTGGACTTACAGAG GAGCCCCATGGGCCGGAAGCAGGGGGGAACGCTGGACGACCTAGAGGAGCAAGCAAGGGAGCTGTACAGGAGACTAAGGGAA AAACCTCGAGACCAGCGAACTGAGGGTGACAGTCAGGAAATGGTACGGCTGCTGCTTCAGGCAATTCAGAGCTTCGAGAAGA AAGTGCGAGTGATCTATACGCAGCTCAGTAAAACTGTGGTTTGCAAGCAGAAGGCGCTGGAACTGTTGCCCAAGGTGGAAGA GGTGGTGAGCTTAATGAATGAGGATGAGAAGACTGTTGTCCGGCTGCAGGAGAAGCGGCAGAAGGAGCTCTGGAATCTCCTG AAGATTGCTTGTAGCAAGGTCCGTGGTCCTGTCAGTGGAAGCCCGGATAGCATGAATGCCTCTCGACTTAGCCAGCCTGGGC AGCTGATGTCTCAGCCCTCCACGGCCTCCAACAGCTTACCTGAGCCAGCCAAGAAGAGTGAAGAACTGGTGGCTGAAGCACA TAACCTCTGCACCCTGCTAGAAAATGCCATACAGGACACTGTGAGGGAACAAGACCAGAGTTTCACGGCCCTAGACTGGAGC TGGTTACAGACGGAAGAAGAAGAGCACAGCTGCCTGGAGCAGGCCTCATGA HumanNF-BCDS (SEQIDNO:29) ATGGCAGAAGATGATCCATATTTGGGAAGGCCTGAACAAATGTTTCATTTGGATCCTTCTTTGACTCATACAATATTTAATC CAGAAGTATTTCAACCACAGATGGCACTGCCAACAGATGGCCCATACCTTCAAATATTAGAGCAACCTAAACAGAGAGGATT TCGTTTCCGTTATGTATGTGAAGGCCCATCCCATGGTGGACTACCTGGTGCCTCTAGTGAAAAGAACAAGAAGTCTTACCCT CAGGTCAAAATCTGCAACTATGTGGGACCAGCAAAGGTTATTGTTCAGTTGGTCACAAATGGAAAAAATATCCACCTGCATG CCCACAGCCTGGTGGGAAAACACTGTGAGGATGGGATCTGCACTGTAACTGCTGGACCCAAGGACATGGTGGTCGGCTTCGC AAACCTGGGTATACTTCATGTGACAAAGAAAAAAGTATTTGAAACACTGGAAGCACGAATGACAGAGGCGTGTATAAGGGGC TATAATCCTGGACTCTTGGTGCACCCTGACCTTGCCTATTTGCAAGCAGAAGGTGGAGGGGACCGGCAGCTGGGAGATCGGG AAAAAGAGCTAATCCGCCAAGCAGCTCTGCAGCAGACCAAGGAGATGGACCTCAGCGTGGTGCGGCTCATGTTTACAGCTTT TCTTCCGGATAGCACTGGCAGCTTCACAAGGCGCCTGGAACCCGTGGTATCAGACGCCATCTATGACAGTAAAGCCCCCAAT GCATCCAACTTGAAAATTGTAAGAATGGACAGGACAGCTGGATGTGTGACTGGAGGGGAGGAAATTTATCTTCTTTGTGACA AAGTTCAGAAAGATGACATCCAGATTCGATTTTATGAAGAGGAAGAAAATGGTGGAGTCTGGGAAGGATTTGGAGATTTTTC CCCCACAGATGTTCATAGACAATTTGCCATTGTCTTCAAAACTCCAAAGTATAAAGATATTAATATTACAAAACCAGCCTCT GTGTTTGTCCAGCTTCGGAGGAAATCTGACTTGGAAACTAGTGAACCAAAACCTTTCCTCTACTATCCTGAAATCAAAGATA AAGAAGAAGTGCAGAGGAAACGTCAGAAGCTCATGCCCAATTTTTCGGATAGTTTCGGCGGTGGTAGTGGTGCTGGAGCTGG AGGCGGAGGCATGTTTGGTAGTGGCGGTGGAGGAGGGGGCACTGGAAGTACAGGTCCAGGGTATAGCTTCCCACACTATGGA TTTCCTACTTATGGTGGGATTACTTTCCATCCTGGAACTACTAAATCTAATGCTGGGATGAAGCATGGAACCATGGACACTG AATCTAAAAAGGACCCTGAAGGTTGTGACAAAAGTGATGACAAAAACACTGTAAACCTCTTTGGGAAAGTTATTGAAACCAC AGAGCAAGATCAGGAGCCCAGCGAGGCCACCGTTGGGAATGGTGAGGTCACTCTAACGTATGCAACAGGAACAAAAGAAGAG AGTGCTGGAGTTCAGGATAACCTCTTTCTAGAGAAGGCTATGCAGCTTGCAAAGAGGCATGCCAATGCCCTTTTCGACTACG CGGTGACAGGAGACGTGAAGATGCTGCTGGCCGTCCAGCGCCATCTCACTGCTGTGCAGGATGAGAATGGGGACAGTGTCTT ACACTTAGCAATCATCCACCTTCATTCTCAACTTGTGAGGGATCTACTAGAAGTCACATCTGGTTTGATTTCTGATGACATT ATCAACATGAGAAATGATCTGTACCAGACGCCCTTGCACTTGGCAGTGATCACTAAGCAGGAAGATGTGGTGGAGGATTTGC TGAGGGCTGGGGCCGACCTGAGCCTTCTGGACCGCTTGGGTAACTCTGTTTTGCACCTAGCTGCCAAAGAAGGACATGATAA AGTTCTCAGTATCTTACTCAAGCACAAAAAGGCAGCACTACTTCTTGACCACCCCAACGGGGACGGTCTGAATGCCATTCAT CTAGCCATGATGAGCAATAGCCTGCCATGTTTGCTGCTGCTGGTGGCCGCTGGGGCTGACGTCAATGCTCAGGAGCAGAAGT CCGGGCGCACAGCACTGCACCTGGCTGTGGAGCACGACAACATCTCATTGGCAGGCTGCCTGCTCCTGGAGGGTGATGCCCA TGTGGACAGTACTACCTACGATGGAACCACACCCCTGCATATAGCAGCTGGGAGAGGGTCCACCAGGCTGGCAGCTCTTCTC AAAGCAGCAGGAGCAGATCCCCTGGTGGAGAACTTTGAGCCTCTCTATGACCTGGATGACTCTTGGGAAAATGCAGGAGAGG ATGAAGGAGTTGTGCCTGGAACCACGCCTCTAGATATGGCCACCAGCTGGCAGGTATTTGACATATTAAATGGGAAACCATA TGAGCCAGAGTTTACATCTGATGATTTACTAGCACAAGGAGACATGAAACAGCTGGCTGAAGATGTGAAGCTGCAGCTGTAT AAGTTACTAGAAATTCCTGATCCAGACAAAAACTGGGCTACTCTGGCGCAGAAATTAGGTCTGGGGATACTTAATAATGCCT TCCGGCTGAGTCCTGCTCCTTCCAAAACACTTATGGACAACTATGAGGTCTCTGGGGGTACAGTCAGAGAGCTGGTGGAGGC CCTGAGACAAATGGGCTACACCGAAGCAATTGAAGTGATCCAGGCAGCCTCCAGCCCAGTGAAGACCACCTCTCAGGCCCAC TCGCTGCCTCTCTCGCCTGCCTCCACAAGGCAGCAAATAGACGAGCTCCGAGACAGTGACAGTGTCTGCGACAGCGGCGTGG AGACATCCTTCCGCAAACTCAGCTTTACCGAGTCTCTGACCAGTGGTGCCTCACTGCTAACTCTCAACAAAATGCCCCATGA TTATGGGCAGGAAGGACCTCTAGAAGGCAAAATTTAG HumanCD14CDS (SEQIDNO:30) ATGGAGCGCGCGTCCTGCTTGTTGCTGCTGCTGCTGCCGCTGGTGCACGTCTCTGCGACCACGCCAGAACCTTGTGAGCTGG ACGATGAAGATTTCCGCTGCGTCTGCAACTTCTCCGAACCTCAGCCCGACTGGTCCGAAGCCTTCCAGTGTGTGTCTGCAGT AGAGGTGGAGATCCATGCCGGCGGTCTCAACCTAGAGCCGTTTCTAAAGCGCGTCGATGCGGACGCCGACCCGCGGCAGTAT GCTGACACGGTCAAGGCTCTCCGCGTGCGGCGGCTCACAGTGGGAGCCGCACAGGTTCCTGCTCAGCTACTGGTAGGCGCCC TGCGTGTGCTAGCGTACTCCCGCCTCAAGGAACTGACGCTCGAGGACCTAAAGATAACCGGCACCATGCCTCCGCTGCCTCT GGAAGCCACAGGACTTGCACTTTCCAGCTTGCGCCTACGCAACGTGTCGTGGGCGACAGGGCGTTCTTGGCTCGCCGAGCTG CAGCAGTGGCTCAAGCCAGGCCTCAAGGTACTGAGCATTGCCCAAGCACACTCGCCTGCCTTTTCCTGCGAACAGGTTCGCG CCTTCCCGGCCCTTACCAGCCTAGACCTGTCTGACAATCCTGGACTGGGCGAACGCGGACTGATGGCGGCTCTCTGTCCCCA CAAGTTCCCGGCCATCCAGAATCTAGCGCTGCGCAACACAGGAATGGAGACGCCCACAGGCGTGTGCGCCGCACTGGCGGCG GCAGGTGTGCAGCCCCACAGCCTAGACCTCAGCCACAACTCGCTGCGCGCCACCGTAAACCCTAGCGCTCCGAGATGCATGT GGTCCAGCGCCCTGAACTCCCTCAATCTGTCGTTCGCTGGGCTGGAACAGGTGCCTAAAGGACTGCCAGCCAAGCTCAGAGT GCTCGATCTCAGCTGCAACAGACTGAACAGGGCGCCGCAGCCTGACGAGCTGCCCGAGGTGGATAACCTGACACTGGACGGG AATCCCTTCCTGGTCCCTGGAACTGCCCTCCCCCACGAGGGCTCAATGAACTCCGGCGTGGTCCCAGCCTGTGCACGTTCGA CCCTGTCGGTGGGGGTGTCGGGAACCCTGGTGCTGCTCCAAGGGGCCCGGGGCTTTGCCTAA HumanMyD88CDS (SEQIDNO:31) ATGCGACCCGACCGCGCTGAGGCTCCAGGACCGCCCGCCATGGCTGCAGGAGGTCCCGGCGCGGGGTCTGCGGCCCCGGTCT CCTCCACATCCTCCCTTCCCCTGGCTGCTCTCAACATGCGAGTGCGGCGCCGCCTGTCTCTGTTCTTGAACGTGCGGACACA GGTGGCGGCCGACTGGACCGCGCTGGCGGAGGAGATGGACTTTGAGTACTTGGAGATCCGGCAACTGGAGACACAAGCGGAC CCCACTGGCAGGCTGCTGGACGCCTGGCAGGGACGCCCTGGCGCCTCTGTAGGCCGACTGCTCGAGCTGCTTACCAAGCTGG GCCGCGACGACGTGCTGCTGGAGCTGGGACCCAGCATTGGTGCCGCCGGATGGTGGTGGTTGTCTCTGATGATTACCTGCAG AGCAAGGAATGTGACTTCCAGACCAAATTTGCACTCAGCCTCTCTCCAGGTGCCCATCAGAAGCGACTGA HumanIRAKCDS (SEQIDNO:32) ATGGCCGGGGGGCCGGGCCCGGGGGAGCCCGCAGCCCCCGGCGCCCAGCACTTCTTGTACGAGGTGCCGCCCTGGGTCATGT GCCGCTTCTACAAAGTGATGGACGCCCTGGAGCCCGCCGACTGGTGCCAGTTCGCCGCCCTGATCGTGCGCGACCAGACCGA GCTGCGGCTGTGCGAGCGCTCCGGGCAGCGCACGGCCAGCGTCCTGTGGCCCTGGATCAACCGCAACGCCCGTGTGGCCGAC CTCGTGCACATCCTCACGCACCTGCAGCTGCTCCGTGCGCGGGACATCATCACAGCCTGGCACCCTCCCGCCCCGCTTCCGT CCCCAGGCACCACTGCCCCGAGGCCCAGCAGCATCCCTGCACCCGCCGAGGCCGAGGCCTGGAGCCCCCGGAAGTTGCCATC CTCAGCCTCCACCTTCCTCTCCCCAGCTTTTCCAGGCTCCCAGACCCATTCAGGGCCTGAGCTCGGCCTGGTCCCAAGCCCT GCTTCCCTGTGGCCTCCACCGCCATCTCCAGCCCCTTCTTCTACCAAGCCAGGCCCAGAGAGCTCAGTGTCCCTCCTGCAGG GAGCCCGCCCCTTTCCGTTTTGCTGGCCCCTCTGTGAGATTTCCCGGGGCACCCACAACTTCTCGGAGGAGCTCAAGATCGG GGAGGGTGGCTTTGGGTGCGTGTACCGGGCGGTGATGAGGAACACGGTGTATGCTGTGAAGAGGCTGAAGGAGAACGCTGAC CTGGAGTGGACTGCAGTGAAGCAGAGCTTCCTGACCGAGGTGGAGCAGCTGTCCAGGTTTCGTCACCCAAACATTGTGGACT TTGCTGGCTACTGTGCTCAGAACGGCTTCTACTGCCTGGTGTACGGCTTCCTGCCCAACGGCTCCCTGGAGGACCGTCTCCA CTGCCAGACCCAGGCCTGCCCACCTCTCTCCTGGCCTCAGCGACTGGACATCCTTCTGGGTACAGCCCGGGCAATTCAGTTT CTACATCAGGACAGCCCCAGCCTCATCCATGGAGACATCAAGAGTTCCAACGTCCTTCTGGATGAGAGGCTGACACCCAAGC TGGGAGACTTTGGCCTGGCCCGGTTCAGCCGCTTTGCCGGGTCCAGCCCCAGCCAGAGCAGCATGGTGGCCCGGACACAGAC AGTGCGGGGCACCCTGGCCTACCTGCCCGAGGAGTACATCAAGACGGGAAGGCTGGCTGTGGACACGGACACCTTCAGCTTT GGGGTGGTAGTGCTAGAGACCTTGGCTGGTCAGAGGGCTGTGAAGACGCACGGTGCCAGGACCAAGTATCTGAAAGACCTGG TGGAAGAGGAGGCTGAGGAGGCTGGAGTGGCTTTGAGAAGCACCCAGAGCACACTGCAAGCAGGTCTGGCTGCAGATGCCTG GGCTGCTCCCATCGCCATGCAGATCTACAAGAAGCACCTGGACCCCAGGCCCGGGCCCTGCCCACCTGAGCTGGGCCTGGGC CTGGGCCAGCTGGCCTGCTGCTGCCTGCACCGCCGGGCCAAAAGGAGGCCTCCTATGACCCAGGAGAACTCCTACGTGTCCA GCACTGGCAGAGCCCACAGTGGGGCTGCTCCATGGCAGCCCCTGGCAGCGCCATCAGGAGCCAGTGCCCAGGCAGCAGAGCA GCTGCAGAGAGGCCCCAACCAGCCCGTGGAGAGTGACGAGAGCCTAGGCGGCCTCTCTGCTGCCCTGCGCTCCTGGCACTTG ACTCCAAGCTGCCCTCTGGACCCAGCACCCCTCAGGGAGGCCGGCTGTCCTCAGGGGGACACGGCAGGAGAATCGAGCTGGG GGAGTGGCCCAGGATCCCGGCCCACAGCCGTGGAAGGACTGGCCCTTGGCAGCTCTGCATCATCGTCGTCAGAGCCACCGCA GATTATCATCAACCCTGCCCGACAGAAGATGGTCCAGAAGCTGGCCCTGTACGAGGATGGGGCCCTGGACAGCCTGCAGCTG CTGTCGTCCAGCTCCCTCCCAGGCTTGGGCCTGGAACAGGACAGGCAGGGGCCCGAAGAAAGTGATGAATTTCAGAGCTGA HumanLBPCDS (SEQIDNO:33) ATGGGGGCCTTGGCCAGAGCCCTGCCGTCCATACTGCTGGCATTGCTGCTTACGTCCACCCCAGAGGCTCTGGGTGCCAACC CCGGCTTGGTCGCCAGGATCACCGACAAGGGACTGCAGTATGCGGCCCAGGAGGGGCTATTAGCTCTGCAGAGTGAGCTGCT CAGGATCACGCTGCCTGACTTCACCGGGGACTTGAGGATCCCCCACGTCGGCCGTGGGCGCTATGAGTTCCACAGCCTGAAC ATCCACAGCTGTGAGCTGCTTCACTCTGCGCTGAGGCCTGTCCCTGGCCAGGGCCTGAGTCTCAGCATCTCCGACTCCTCCA TCCGGGTCCAGGGCAGGTGGAAGGTGCGCAAGTCATTCTTCAAACTACAGGGCTCCTTTGATGTCAGTGTCAAGGGCATCAG CATTTCGGTCAACCTCCTGTTGGGCAGCGAGTCCTCCGGGAGGCCCACAGTTACTGCCTCCAGCTGCAGCAGTGACATCGCT GACGTGGAGGTGGACATGTCGGGAGACTTGGGGTGGCTGTTGAACCTCTTCCACAACCAGATTGAGTCCAAGTTCCAGAAAG TACTGGAGAGCAGGATTTGCGAAATGATCCAGAAATCAGTGTCCTCCGATCTACAGCCTTATCTCCAAACTCTGCCAGTTAC AACAGAGATTGACAGTTTCGCCGACATTGATTATAGCTTAGTGGAAGCCCCTCGGGCAACAGCCCAGATGCTGGAGGTGATG TTTAAGGGTGAAATCTTTCATCGTAACCACCGTTCTCCAGTTACCCTCCTTGCTGCAGTCATGAGCCTTCCTGAGGAACACA ACAAAATGGTCTACTTTGCCATCTCGGATTATGTCTTCAACACGGCCAGCCTGGTTTATCATGAGGAAGGATATCTGAACTT CTCCATCACAGATGACATGATACCGCCTGACTCTAATATCCGACTGACCACCAAGTCCTTCCGACCCTTCGTCCCACGGTTA GCCAGGCTCTACCCCAACATGAACCTGGAACTCCAGGGATCAGTGCCCTCTGCTCCGCTCCTGAACTTCAGCCCTGGGAATC TGTCTGTGGACCCCTATATGGAGATAGATGCCTTTGTGCTCCTGCCCAGCTCCAGCAAGGAGCCTGTCTTCCGGCTCAGTGT GGCCACTAATGTGTCCGCCACCTTGACCTTCAATACCAGCAAGATCACTGGGTTCCTGAAGCCAGGAAAGGTAAAAGTGGAA CTGAAAGAATCCAAAGTTGGACTATTCAATGCAGAGCTGTTGGAAGCGCTCCTCAACTATTACATCCTTAACACCTTCTACC CCAAGTTCAATGATAAGTTGGCCGAAGGCTTCCCCCTTCCTCTGCTGAAGCGTGTTCAGCTCTACGACCTTGGGCTGCAGAT CCATAAGGACTTCCTGTTCTTGGGTGCCAATGTCCAATACATGAGAGTTTGA HumanTRAF6CDS (SEQIDNO:34) ATGAGTCTGCTAAACTGTGAAAACAGCTGTGGATCCAGCCAGTCTGAAAGTGACTGCTGTGTGGCCATGGCCAGCTCCTGTA GCGCTGTAACAAAAGATGATAGTGTGGGTGGAACTGCCAGCACGGGGAACCTCTCCAGCTCATTTATGGAGGAGATCCAGGG ATATGATGTAGAGTTTGACCCACCCCTGGAAAGCAAGTATGAATGCCCCATCTGCTTGATGGCATTACGAGAAGCAGTGCAA ACGCCATGCGGCCATAGGTTCTGCAAAGCCTGCATCATAAAATCAATAAGGGATGCAGGTCACAAATGTCCAGTTGACAATG AAATACTGCTGGAAAATCAACTATTTCCAGACAATTTTGCAAAACGTGAGATTCTTTCTCTGATGGTGAAATGTCCAAATGA AGGTTGTTTGCACAAGATGGAACTGAGACATCTTGAGGATCATCAAGCACATTGTGAGTTTGCTCTTATGGATTGTCCCCAA TGCCAGCGTCCCTTCCAAAAATTCCATATTAATATTCACATTCTGAAGGATTGTCCAAGGAGACAGGTTTCTTGTGACAACT GTGCTGCATCAATGGCATTTGAAGATAAAGAGATCCATGACCAGAACTGTCCTTTGGCAAATGTCATCTGTGAATACTGCAA TACTATACTCATCAGAGAACAGATGCCTAATCATTATGATCTAGACTGCCCTACAGCCCCAATTCCATGCACATTCAGTACT TTTGGTTGCCATGAAAAGATGCAGAGGAATCACTTGGCACGCCACCTACAAGAGAACACCCAGTCACACATGAGAATGTTGG CCCAGGCTGTTCATAGTTTGAGCGTTATACCCGACTCTGGGTATATCTCAGAGGTCCGGAATTTCCAGGAAACTATTCACCA GTTAGAGGGTCGCCTTGTAAGACAAGACCATCAAATCCGGGAGCTGACTGCTAAAATGGAAACTCAGAGTATGTATGTAAGT GAGCTCAAACGAACCATTCGAACCCTTGAGGACAAAGTTGCTGAAATCGAAGCACAGCAGTGCAATGGAATTTATATTTGGA AGATTGGCAACTTTGGAATGCATTTGAAATGTCAAGAAGAGGAGAAACCTGTTGTGATTCATAGCCCTGGATTCTACACTGG CAAACCCGGGTACAAACTGTGCATGCGCTTGCACCTTCAGTTACCGACTGCTCAGCGCTGTGCAAACTATATATCCCTTTTT GTCCACACAATGCAAGGAGAATATGACAGCCACCTCCCTTGGCCCTTCCAGGGTACAATACGCCTTACAATTCTTGATCAGT CTGAAGCACCTGTAAGGCAAAACCACGAAGAGATAATGGATGCCAAACCAGAGCTGCTTGCTTTCCAGCGACCCACAATCCC ACGGAACCCAAAAGGTTTTGGCTATGTAACTTTTATGCATCTGGAAGCCCTAAGACAAAGAACTTTCATTAAGGATGACACA TTATTAGTGCGCTGTGAGGTCTCCACCCGCTTTGACATGGGTAGCCTTCGGAGGGAGGGTTTTCAGCCACGAAGTACTGATG CAGGGGTATAG HumanK-RasCDS (SEQIDNO:35) ATGACTGAATATAAACTTGTGGTAGTTGGAGCTGGTGGCGTAGGCAAGAGTGCCTTGACGATACAGCTAATTCAGAATCATT TTGTGGACGAATATGATCCAACAATAGAGGATTCCTACAGGAAGCAAGTAGTAATTGATGGAGAAACCTGTCTCTTGGATAT TCTCGACACAGCAGGTCAAGAGGAGTACAGTGCAATGAGGGACCAGTACATGAGGACTGGGGAGGGCTTTCTTTGTGTATTT GCCATAAATAATACTAAATCATTTGAAGATATTCACCATTATAGAGAACAAATTAAAAGAGTTAAGGACTCTGAAGATGTAC CTATGGTCCTAGTAGGAAATAAATGTGATTTGCCTTCTAGAACAGTAGACACAAAACAGGCTCAGGACTTAGCAAGAAGTTA TGGAATTCCTTTTATTGAAACATCAGCAAAGACAAGACAGGGTGTTGATGATGCCTTCTATACATTAGTTCGAGAAATTCGA AAACATAAAGAAAAGATGAGCAAAGATGGTAAAAAGAAGAAAAAGAAGTCAAAGACAAAGTGTGTAATTATGTAA HumanN-RasCDS (SEQIDNO:36) ATGACTGAGTACAAACTGGTGGTGGTTGGAGCAGGTGGTGTTGGGAAAAGCGCACTGACAATCCAGCTAATCCAGAACCACT TTGTAGATGAATATGATCCCACCATAGAGGATTCTTACAGAAAACAAGTGGTTATAGATGGTGAAACCTGTTTGTTGGACAT ACTGGATACAGCTGGACAAGAAGAGTACAGTGCCATGAGAGACCAATACATGAGGACAGGCGAAGGCTTCCTCTGTGTATTT GCCATCAATAATAGCAAGTCATTTGCGGATATTAACCTCTACAGGGAGCAGATTAAGCGAGTAAAAGACTCGGATGATGTAC CTATGGTGCTAGTGGGAAACAAGTGTGATTTGCCAACAAGGACAGTTGATACAAAACAAGCCCACGAACTGGCCAAGAGTTA CGGGATTCCATTCATTGAAACCTCAGCCAAGACCAGACAGGGTGTTGAAGATGCTTTTTACACACTGGTAAGAGAAATACGC CAGTACCGAATGAAAAAACTCAACAGCAGTGATGATGGGACTCAGGGTTGTATGGGATTGCCATGTGTGGTGATGTAA HumanRafCDS (SEQIDNO:37) ATGGCTAGCAAACGAAAATCTACAACTCCATGCATGGTTCGGACATCACAAGTAGTAGAACAAGATGTGCCCGAGGAAGTAG ACAGGGCCAAAGAGAAAGGAATCGGCACACCACAGCCTGACGTGGCCAAGGACAGTTGGGCAGCAGAACTTGAAAACTCTTC CAAAGAAAACGAAGTGATAGAGGTGAAATCTATGGGGGAAAGCCAGTCCAAAAAACTCCAAGGTGGTTATGAGTGCAAATAC TGCCCCTACTCCACGCAAAACCTGAACGAGTTCACGGAGCATGTCGACATGCAGCATCCCAACGTGATTCTCAACCCCCTCT ACGTGTGTGCAGAATGTAACTTCACAACCAAAAAGTACGACTCCCTATCCGACCACAACTCCAAGTTCCATCCCGGGGAGGC CAACTTCAAGCTGAAGTTAATTAAACGCAATAATCAAACTGTCTTGGAACAGTCCATCGAAACCACCAACCATGTCGTGTCC ATCACCACCAGTGGCCCTGGAACTGGTGACAGTGATTCTGGGATCTCGGTGAGTAAAACCCCCATCATGAAGCCTGGAAAAC CAAAAGCGGATGCCAAGAAGGTGCCCAAGAAGCCCGAGGAGATCACCCCCGAGAACCACGTGGAAGGGACCGCCCGCCTGGT GACAGACACAGCTGAGATCCTCTCGAGACTCGGCGGGGTGGAGCTCCTCCAAGACACATTAGGACACGTCATGCCTTCTGTA CAGCTGCCACCAAATATCAACCTTGTGCCCAAGGTCCCTGTCCCACTAAATACTACCAAATACAACTCTGCCCTGGATACAA ATGCCACGATGATCAACTCTTTCAACAAGTTTCCTTACCCGACCCAGGCTGAGTTGTCCTGGCTGACAGCTGCCTCCAAACA CCCAGAGGAGCACATCAGAATCTGGTTTGCCACCCAGCGCTTAAAGCATGGCATCAGCTGGTCCCCAGAAGAGGTGGAGGAG GCCCGGAAGAAGATGTTCAACGGCACCATCCAGTCAGTACCCCCGACCATCACTGTGCTGCCCGCCCAGTTGGCCCCCACAA AGGTGACGCAGCCCATCCTCCAGACGGCTCTACCGTGCCAGATCCTCGGCCAGACTAGCCTGGTGCTGACTCAGGTGACCAG CGGGTCAACAACCGTCTCTTGCTCCCCCATCACACTTGCCGTGGCAGGAGTCACCAACCATGGCCAGAAGAGACCCTTGGTG ACTCCCCAAGCTGCCCCCGAACCCAAGCGTCCACACATCGCTCAGGTGCCAGAGCCCCCACCCAAGGTGGCCAACCCCCCGC TCACACCAGCCAGTGACCGCAAGAAGACAAAGGAGCAGATAGCACATCTCAAGGCCAGCTTTCTCCAGAGCCAGTTCCCTGA CGATGCCGAGGTTTACCGGCTCATCGAGGTGACTGGCCTTGCCAGGAGCGAGATCAAGAAGTGGTTCAGTGACCACCGATAT CGGTGTCAAAGGGGCATCGTCCACATCACCAGCGAATCCCTTGCCAAAGACCAGTTGGCCATCGCGGCCTCCCGACACGGTC GCACGTATCATGCGTACCCAGACTTTGCCCCCCAGAAGTTCAAAGAGAAAACACAGGGTCAGGTTAAAATCTTGGAAGACAG CTTTTTGAAAAGTTCTTTTCCTACCCAAGCAGAACTGGATCGGCTAAGGGTGGAGACCAAGCTGAGCAGGAGAGAGATCGAC TCCTGGTTCTCGGAGAGGCGGAAGCTTCGAGACAGCATGGAACAAGCTGTCTTGGATTCCATGGGGTCTGGCAAAAAAGGCC AAGATGTGGGAGCCCCCAATGGTGCTCTGTCTCGACTCGACCAGCTCTCCGGTGCCCAGTTAACAAGTTCTCTGCCCAGCCC TTCGCCAGCAATTGCAAAAAGTCAAGAACAGGTTCATCTCCTGAGGAGCACGTTTGCAAGAACCCAGTGGCCTACTCCCCAG GAGTACGACCAGTTAGCGGCCAAGACTGGCCTGGTCCGAACTGAGATTGTGCGTTGGTTCAAGGAGAACAGATGCTTGCTGA AAACGGGAACCGTGAAGTGGATGGAGCAGTACCAGCACCAGCCCATGGCAGATGATCACGGCTACGATGCCGTAGCAAGGAA AGCAACAAAACCCATGGCCGAGAGCCCAAAGAACGGGGGTGATGTGGTTCCACAATATTACAAGGACCCCAAAAAGCTCTGC GAAGAGGACTTGGAGAAGTTGGTGACCAGGGTAAAAGTAGGCAGCGAGCCAGCAAAAGACTGTTTGCCAGCAAAGCCCTCAG AGGCCACCTCAGACCGGTCAGAGGGCAGCAGCCGGGACGGCCAGGGTAGCGACGAGAACGAGGAGTCGAGCGTTGTGGATTA CGTGGAGGTGACGGTCGGGGAGGAGGATGCGATCTCAGATAGATCAGATAGCTGGAGTCAGGCTGCGGCAGAAGGTGTGTCG GAACTGGCTGAATCAGACTCCGACTGCGTCCCTGCAGAGGCTGGCCAGGCCTAG HumanNMK1CDS (SEQIDNO:38) ATGCCCAAGAAGAAGCCGACGCCCATCCAGCTGAACCCGGCCCCCGACGGCTCTGCAGTTAACGGGACCAGCTCTGCGGAGA CCAACTTGGAGGCCTTGCAGAAGAAGCTGGAGGAGCTAGAGCTTGATGAGCAGCAGCGAAAGCGCCTTGAGGCCTTTCTTAC CCAGAAGCAGAAGGTGGGAGAACTGAAGGATGACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTG TTCAAGGTCTCCCACAAGCCTTCTGGCCTGGTCATGGCCAGAAAGCTAATTCATCTGGAGATCAAACCCGCAATCCGGAACC AGATCATAAGGGAGCTGCAGGTTCTGCATGAGTGCAACTCTCCGTACATCGTGGGCTTCTATGGTGCGTTCTACAGCGATGG CGAGATCAGTATCTGCATGGAGCACATGGATGGAGGTTCTCTGGATCAAGTCCTGAAGAAAGCTGGAAGAATTCCTGAACAA ATTTTAGGAAAAGTTAGCATTGCTGTAATAAAAGGCCTGACATATCTGAGGGAGAAGCACAAGATCATGCACAGAGATGTCA AGCCCTCCAACATCCTAGTCAACTCCCGTGGGGAGATCAAGCTCTGTGACTTTGGGGTCAGCGGGCAGCTCATCGACTCCAT GGCCAACTCCTTCGTGGGCACAAGGTCCTACATGTCGCCAGAAAGACTCCAGGGGACTCATTACTCTGTGCAGTCAGACATC TGGAGCATGGGACTGTCTCTGGTAGAGATGGCGGTTGGGAGGTATCCCATCCCTCCTCCAGATGCCAAGGAGCTGGAGCTGA TGTTTGGGTGCCAGGTGGAAGGAGATGCGGCTGAGACCCCACCCAGGCCAAGGACCCCCGGGAGGCCCCTTAGCTCATACGG AATGGACAGCCGACCTCCCATGGCAATTTTTGAGTTGTTGGATTACATAGTCAACGAGCCTCCTCCAAAACTGCCCAGTGGA GTGTTCAGTCTGGAATTTCAAGATTTTGTGAATAAATGCTTAATAAAAAACCCCGCAGAGAGAGCAGATTTGAAGCAACTCA TGGTTCATGCTTTTATCAAGAGATCTGATGCTGAGGAAGTGGATTTTGCAGGTTGGCTCTGCTCCACCATCGGCCTTAACCA GCCCAGCACACCAACCCATGCTGCTGGCGTCTAA HumanNMK2CADS (SEQIDNO:39) ATGCTGGCCCGGAGGAAGCCGGTGCTGCCGGCGCTCACCATCAACCCTACCATCGCCGAGGGCCCATCCCCTACCAGCGAGG GCGCCTCCGAGGCAAACCTGGTGGACCTGCAGAAGAAGCTGGAGGAGCTGGAACTTGACGAGCAGCAGAAGAAGCGGCTGGA AGCCTTTCTCACCCAGAAAGCCAAGGTCGGCGAACTCAAAGACGATGACTTCGAAAGGATCTCAGAGCTGGGCGCGGGCAAC GGCGGGGTGGTCACCAAAGTCCAGCACAGACCCTCGGGCCTCATCATGGCCAGGAAGCTGATCCACCTTGAGATCAAGCCGG CCATCCGGAACCAGATCATCCGCGAGCTGCAGGTCCTGCACGAATGCAACTCGCCGTACATCGTGGGCTTCTACGGGGCCTT CTACAGTGACGGGGAGATCAGCATTTGCATGGAACACATGGACGGCGGCTCCCTGGACCAGGTGCTGAAAGAGGCCAAGAGG ATTCCCGAGGAGATCCTGGGGAAAGTCAGCATCGCGGTTCTCCGGGGCTTGGCGTACCTCCGAGAGAAGCACCAGATCATGC ACCGAGATGTGAAGCCCTCCAACATCCTCGTGAACTCTAGAGGGGAGATCAAGCTGTGTGACTTCGGGGTGAGCGGCCAGCT CATCGACTCCATGGCCAACTCCTTCGTGGGCACGCGCTCCTACATGGCTCCGGAGCGGTTGCAGGGCACACATTACTCGGTG CAGTCGGACATCTGGAGCATGGGCCTGTCCCTGGTGGAGCTGGCCGTCGGAAGGTACCCCATCCCCCCGCCCGACGCCAAAG AGCTGGAGGCCATCTTTGGCCGGCCCGTGGTCGACGGGGAAGAAGGAGAGCCTCACAGCATCTCGCCTCGGCCGAGGCCCCC CGGGCGCCCCGTCAGCGGTCACGGGATGGATAGCCGGCCTGCCATGGCCATCTTTGAACTCCTGGACMCFATTGTGAACGAG CCACCTCCTAAGCTGCCCAACGGTGTGTTCACCCCCGACTTCCAGGAGTTTGTCAATAAATGCCTCATCAAGAACCCAGCGG AGCGGGCGGACCTGAAGATGCTCACAAACCACACCTTCATCAAGCGGTCCGAGGTGGAAGAAGTGGATTTTGCCGGCTGGTT GTGTAAAACCCTGCGGCTGAACCAGCCCGGCACACCCACGCGCACCGCCGTGTGA HumanERK1CDS (SEQIDNO:40) ATGGCGGCGGCGGCGGCTCAGGGGGGCGGGGGCGGGGAGCCCCGTAGAACCGAGGGGGTCGGCCCGGGGGTCCCGGGGGAGG TGGAGATGGTGAAGGGGCAGCCGTTCGACGTGGGCCCGCGCTACACGCAGTTGCAGTACATCGGCGAGGGCGCGTACGGCAT GGTCAGCTCGGCCTATGACCACGTGCGCAAGACTCGCGTGGCCATCAAGAAGATCAGCCCCTTCGAACATCAGACCTACTGC CAGCGCACGCTCCGGGAGATCCAGATCCTGCTGCGCTTCCGCCATGAGAATGTCATCGGCATCCGAGACATTCTGCGGGCGT CCACCCTGGAAGCCATGAGAGATGTCTACATTGTGCAGGACCTGATGGAGACTGACCTGTACAAGTTGCTGAAAAGCCAGCA GCTGAGCAATGACCATATCTGCTACTTCCTCTACCAGATCCTGCGGGGCCTCAAGTACATCCACTCCGCCAACGTGCTCCAC CGAGATCTAAAGCCCTCCAACCTGCTCATCAACACCACCTGCGACCTTAAGATTTGTGATTTCGGCCTGGCCCGGATTGCCG ATCCTGAGCATGACCACACCGGCTTCCTGACGGAGTATGTGGCTACGCGCTGGTACCGGGCCCCAGAGATCATGCTGAACTC CAAGGGCTATACCAAGTCCATCGACATCTGGTCTGTGGGCTGCATTCTGGCTGAGATGCTCTCTAACCGGCCCATCTTCCCT GGCAAGCACTACCTGGATCAGCTCAACCACATTCTGGGCATCCTGGGCTCCCCATCCCAGGAGGACCTGAATTGTATCATCA ACATGAAGGCCCGAAACTACCTACAGTCTCTGCCCTCCAAGACCAAGGTGGCTTGGGCCAAGCTTTTCCCCAAGTCAGACTC CAAAGCCCTTGACCTGCTGGACCGGATGTTAACCTTTAACCCCAATAAACGGATCACAGTGGAGGAAGCGCTGGCTCACCCC TACCTGGAGCAGTACTATGACCCGACGGATGAGGTGGGCCAGTCCCCAGCAGCAGTGGGGCTGGGGGCAGGGGAGCAGGGGG GCACGTAG HumanERK2CDS (SEQIDNO:41) ATGGCGGCGGCGGCGGCGGCGGGCGCGGGCCCGGAGATGGTCCGCGGGCAGGTGTTCGACGTGGGGCCGCGCTACACCAACC TCTCGTACATCGGCGAGGGCGCCTACGGCATGGTGTGCTCTGCTTATGATAATGTCAACAAAGTTCGAGTAGCTATCAAGAA AATCAGCCCCTTTGAGCACCAGACCTACTGCCAGAGAACCCTGAGGGAGATAAAAATCTTACTGCGCTTCAGACATGAGAAC ATCATTGGAATCAATGACATTATTCGAGCACCAACCATCGAGCAAATGAAAGATGTATATATAGTACAGGACCTCATGGAAA CAGATCTTTACAAGCTCTTGAAGACACAACACCTCAGCAATGACCATATCTGCTATTTTCTCTACCAGATCCTCAGAGGGTT AAAATATATCCATTCAGCTAACGTTCTGCACCGTGACCTCAAGCCTTCCAACCTGCTGCTCAACACCACCTGTGATCTCAAG ATCTGTGACTTTGGCCTGGCCCGTGTTGCAGATCCAGACCATGATCACACAGGGTTCCTGACAGAATATGTGGCCACACGTT GGTACAGGGCTCCAGAAATTATGTTGAATTCCAAGGGCTACACCAAGTCCATTGATATTTGGTCTGTAGGCTGCATTCTGGC AGAAATGCTTTCTAACAGGCCCATCTTTCCAGGGAAGCATTATCTTGACCAGCTGAACCACATTTTGGGTATTCTTGGATCC CCATCACAAGAAGACCTGAATTGTATAATAAATTTAAAAGCTAGGAACTATTTGCTTTCTCTTCCACACAAAAATAAGGTGC CATGGAACAGGCTGTTCCCAAATGCTGACTCCAAAGCTCTGGACTTATTGGACAAAATGTTGACATTCAACCCACACAAGAG GATTGAAGTAGAACAGGCTCTGGCCCACCCATATCTGGAGCAGTATTACGACCCGAGTGACGAGCCCATCGCCGAAGCACCA TTCAAGTTCGACATGGAATTGGATGACTTGCCTAAGGAAAAGCTCAAAGAACTAATTTTTGAAGAGACTGCTAGATTCCAGC CAGGATACAGATCTTAA HumanIBCDS (SEQIDNO:42) ATGTTCCAGGCGGCCGAGCGCCCCCAGGAGTGGGCCATGGAGGGCCCCCGCGACGGGCTGAAGAAGGAGCGGCTACTGGACG ACCGCCACGACAGCGGCCTGGACTCCATGAAAGACGAGGAGTACGAGCAGATGGTCAAGGAGCTGCAGGAGATCCGCCTCGA GCCGCAGGAGGTGCCGCGCGGCTCGGAGCCCTGGAAGCAGCAGCTCACCGAGGACGGGGACTCGTTCCTGCACTTGGCCATC ATCCATGAAGAAAAGGCACTGACCATGGAAGTGATCCGCCAGGTGAAGGGAGACCTGGCCTTCCTCAACTTCCAGAACAACC TGCAGCAGACTCCACTCCACTTGGCTGTGATCACCAACCAGCCAGAAATTGCTGAGGCACTTCTGGGAGCTGGCTGTGATCC TGAGCTCCGAGACTTTCGAGGAAATACCCCCCTACACCTTGCCTGTGAGCAGGGCTGCCTGGCCAGCGTGGGAGTCCTGACT CAGTCCTGCACCACCCCGCACCTCCACTCCATCCTGAAGGCTACCAACTACAATGGCCACACGTGTCTACACTTAGCCTCTA TCCATGGCTACCTGGGCATCGTGGAGCTTTTGGTGTCCTTGGGTGCTGATGTCAATGCTCAGGAGCCCTGTAATGGCCGGAC TGCCCTTCACCTCGCAGTGGACCTGCAAAATCCTGACCTGGTGTCACTCCTGTTGAAGTGTGGGGCTGATGTCAACAGAGTT ACCTACCAGGGCTATTCTCCCTACCAGCTCACCTGGGGCCGCCCAAGCACCCGGATACAGCAGCAGCTGGGCCAGCTGACAC TAGAAAACCTTCAGATGCTGCCAGAGAGTGAGGATGAGGAGAGCTATGACACAGAGTCAGAGTTCACGGAGTTCACAGAGGA CGAGCTGCCCTATGATGACTGTGTGTTTGGAGGCCAGCGTCTGACGTTATGA HumanRacCDS (SEQIDNO:43) ATGAGCGACGTGGCTATTGTGAAGGAGGGTTGGCTGCACAAACGAGGGGAGTACATCAAGACCTGGCGGCCACGCTACTTCC TCCTCAAGAATGATGGCACCTTCATTGGCTACAAGGAGCGGCCGCAGGATGTGGACCAACGTGAGGCTCCCCTCAACAACTT CTCTGTGGCGCAGTGCCAGCTGATGAAGACGGAGCGGCCCCGGCCCAACACCTTCATCATCCGCTGCCTGCAGTGGACCACT GTCATCGAACGCACCTTCCATGTGGAGACTCCTGAGGAGCGGGAGGAGTGGACAACCGCCATCCAGACTGTGGCTGACGGCC TCAAGAAGCAGGAGGAGGAGGAGATGGACTTCCGGTCGGGCTCACCCAGTGACAACTCAGGGGCTGAAGAGATGGAGGTGTC CCTGGCCAAGCCCAAGCACCGCGTGACCATGAACGAGTTTGAGTACCTGAAGCTGCTGGGCAAGGGCACTTTCGGCAAGGTG ATCCTGGTGAAGGAGAAGGCCACAGGCCGCTACTACGCCATGAAGATCCTCAAGAAGGAAGTCATCGTGGCCAAGGACGAGG TGGCCCACACACTCACCGAGAACCGCGTCCTGCAGAACTCCAGGCACCCCTTCCTCACAGCCCTGAAGTACTCTTTCCAGAC CCACGACCGCCTCTGCTTTGTCATGGAGTACGCCAACGGGGGCGAGCTGTTCTTCCACCTGTCCCGGGAGCGTGTGTTCTCC GAGGACCGGGCCCGCTTCTATGGCGCTGAGATTGTGTCAGCCCTGGACTACCTGCACTCGGAGAAGAACGTGGTGTACCGGG ACCTCAAGCTGGAGAACCTCATGCTGGACAAGGACGGGCACATTAAGATCACAGACTTCGGGCTGTGCAAGGAGGGGATCAA GGACGGTGCCACCATGAAGACCTTTTGCGGCACACCTGAGTACCTGGCCCCCGAGGTGCTGGAGGACAATGACTACGGCCGT GCAGTGGACTGGTGGGGGCTGGGCGTGGTCATGTACGAGATGATGTGCGGTCGCCTGCCCTTCTACAACCAGGACCATGAGA AGCTTTTTGAGCTCATCCTCATGGAGGAGATCCGCTTCCCGCGCACGCTTGGTCCCGAGGCCAAGTCCTTGCTTTCAGGGCT GCTCAAGAAGGACCCCAAGCAGAGGCTTGGCGGGGGCTCCGAGGACGCCAAGGAGATCATGCAGCATCGCTTCTTTGCCGGT ATCGTGTGGCAGCACGTGTACGAGAAGAAGCTCAGCCCACCCTTCAAGCCCCAGGTCACGTCGGAGACTGACACCAGGTATT TTGATGAGGAGTTCACGGCCCAGATGATCACCATCACACCACCTGACCAAGATGACAGCATGGAGTGTGTGGACAGCGAGCG CAGGCCCCACTTCCCCCAGTTCTCCTACTCGGCCAGCGGCACGGCCTGA HumanMEK3CDS (SEQIDNO:44) ATGTCCAAGCCACCCGCACCCAACCCCACACCCCCCCGGAACCTGGACTCCCGGACCTTCATCACCATTGGAGACAGAAACT TTGAGGTGGAGGCTGATGACTTGGTGACCATCTCAGAACTGGGCCGTGGAGCCTATGGGGTGGTAGAGAAGGTGCGGCACGC CCAGAGCGGCACCATCATGGCCGTGAAGCGGATCCGGGCCACCGTGAACTCACAGGAGCAGAAGCGGCTGCTCATGGACCTG GACATCAACATGCGCACGGTCGACTGTTTCTACACTGTCACCTTCTACGGGGCACTATTCAGAGAGGGAGACGTGTGGATCT GCATGGAGCTCATGGACACATCCTTGGACAAGTTCTACCGGAAGGTGCTGGATAAAAACATGACAATTCCAGAGGACATCCT TGGGGAGATTGCTGTGTCTATCGTGCGGGCCCTGGAGCATCTGCACAGCAAGCTGTCGGTGATCCACAGAGATGTGAAGCCC TCCAATGTCCTTATCAACAAGGAGGGCCATGTGAAGATGTGTGACTTTGGCATCAGTGGCTACTTGGTGGACTCTGTGGCCA AGACGATGGATGCCGGCTGCAAGCCCTACATGGCCCCTGAGAGGATCAACCCAGAGCTGAACCAGAAGGGCTACAATGTCAA GTCCGACGTCTGGAGCCTGGGCATCACCATGATTGAGATGGCCATCCTGCGGTTCCCTTACGAGTCCTGGGGGACCCCGTTC CAGCAGCTGAAGCAGGTGGTGGAGGAGCCGTCCCCCCAGCTCCCAGCCGACCGTTTCTCCCCCGAGTTTGTGGACTTCACTG CTCAGTGCCTGAGGAAGAACCCCGCAGAGCGTATGAGCTACCTGGAGCTGATGGAGCACCCCTTCTTCACCTTGCACAAAAC CAAGAAGACGGACATTGCTGCCTTCGTGAAGGAGATCCTGGGAGAAGACTCATAG HumanMEK6CDS (SEQIDNO:45) ATGGAACTGGGACGAGGTGCGTACGGGGTGGTGGAGAAGATGCGGCACGTGCCCAGCGGGCAGATCATGGCAGTGAAGCGGA TCCGAGCCACAGTAAATAGCCAGGAACAGAAACGGCTACTGATGGATTTGGATATTTCCATGAGGACGGTGGACTGTCCATT CACTGTCACCTTTTATGGCGCACTGTTTCGGGAGGGTGATGTGTGGATCTGCATGGAGCTCATGGATACATCACTAGATAAA TTCTACAAACAAGTTATTGATAAAGGCCAGACAATTCCAGAGGACATCTTAGGGAAAATAGCAGTTTCTATTGTAAAAGCAT TAGAACATTTACATAGTAAGCTGTCTGTCATTCACAGAGACGTCAAGCCTTCTAATGTACTCATCAATGCTCTCGGTCAAGT GAAGATGTGCGATTTTGGAATCAGTGGCTACTTGGTGGACTCTGTTGCTAAAACAATTGATGCAGGTTGCAAACCATACATG GCCCCTGAAAGAATAAACCCAGAGCTCAACCAGAAGGGATACAGTGTGAAGTCTGACATTTGGAGTCTGGGCATCACGATGA TTGAGTTGGCCATCCTTCGATTTCCCTATGATTCATGGGGAACTCCATTTCAGCAGCTCAAACAGGTGGTAGAGGAGCCATC GCCACAACTCCCAGCAGACAAGTTCTCTGCAGAGTTTGTTGACTTTACCTCACAGTGCTTAAAGAAGAATTCCAAAGAACGG CCTACATACCCAGAGCTAATGCAACATCCATTTTTCACCCTACATGAATCCAAAGGAACAGATGTGGCATCTTTTGTAAAAC TGATTCTTGGAGACTAA Humanp38CDS (SEQIDNO:46) ATGTCTCAGGAGAGGCCCACGTTCTACCGGCAGGAGCTGAACAAGACAATCTGGGAGGTGCCCGAGCGTTACCAGAACCTGT CTCCAGTGGGCTCTGGCGCCTATGGCTCTGTGTGTGCTGCTTTTGACACAAAAACGGGGTTACGTGTGGCAGTGAAGAAGCT CTCCAGACCATTTCAGTCCATCATTCATGCGAAAAGAACCTACAGAGAACTGCGGTTACTTAAACATATGAAACATGAAAAT GTGATTGGTCTGTTGGACGTTTTTACACCTGCAAGGTCTCTGGAGGAATTCAATGATGTGTATCTGGTGACCCATCTCATGG GGGCAGATCTGAACAACATTGTGAAATGTCAGAAGCTTACAGATGACCATGTTCAGTTCCTTATCTACCAAATTCTCCGAGG TCTAAAGTATATACATTCAGCTGACATAATTCACAGGGACCTAAAACCTAGTAATCTAGCTGTGAATGAAGACTGTGAGCTG AAGATTCTGGATTTTGGACTGGCTCGGCACACAGATGATGAAATGACAGGCTACGTGGCCACTAGGTGGTACAGGGCTCCTG AGATCATGCTGAACTGGATGCATTACAACCAGACAGTTGATATTTGGTCAGTGGGATGCATAATGGCCGAGCTGTTGACTGG AAGAACATTGTTTCCTGGTACAGACCATATTAACCAGCTTCAGCAGATTATGCGTCTGACAGGAACACCCCCCGCTTATCTC ATTAACAGGATGCCAAGCCATGAGGCAAGAAACTATATTCAGTCTTTGACTCAGATGCCGAAGATGAACTTTGCGAATGTAT TTATTGGTGCCAATCCCCTGGCTGTCGACTTGCTGGAGAAGATGCTTGTATTGGACTCAGATAAGAGAATTACAGCGGCCCA AGCCCTTGCACATGCCTACTTTGCTCAGTACCACGATCCTGATGATGAACCAGTGGCCGATCCTTATGATCAGTCCTTTGAA AGCAGGGACCTCCTTATAGATGAGTGGAAAAGCCTGACCTATGATGAAGTCATCAGCTTTGTGCCACCACCCCTTGACCAAG AAGAGATGGAGTCCTGA HumanPKRCDS (SEQIDNO:47) ATGGCTGGTGATCTTTCAGCAGGTTTCTTCATGGAGGAACTTAATACATACCGTCAGAAGCAGGGAGTAGTACTTAAATATC AAGAACTGCCTAATTCAGGACCTCCACATGATAGGAGGTTTACATTTCAAGTTATAATAGATGGAAGAGAATTTCCAGAAGG TGAAGGTAGATCAAAGAAGGAAGCAAAAAATGCCGCAGCCAAATTAGCTGTTGAGATACTTAATAAGGAAAAGAAGGCAGTT AGTCCTTTATTATTGACAACAACGAATTCTTCAGAAGGATTATCCATGGGGAATTACATAGGCCTTATCAATAGAATTGCCC AGAAGAAAAGACTAACTGTAAATTATGAACAGTGTGCATCGGGGGTGCATGGGCCAGAAGGATTTCATTATAAATGCAAAAT GGGACAGAAAGAATATAGTATTGGTACAGGTTCTACTAAACAGGAAGCAAAACAATTGGCCGCTAAACTTGCATATCTTCAG ATATTATCAGAAGAAACCTCAGTGAAATCTGACTACCTGTCCTCTGGTTCTTTTGCTACTACGTGTGAGTCCCAAAGCAACT CTTTAGTGACCAGCACACTCGCTTCTGAATCATCATCTGAAGGTGACTTCTCAGCAGATACATCAGAGATAAATTCTAACAG TGACAGTTTAAACAGTTCTTCGTTGCTTATGAATGGTCTCAGAAATAATCAAAGGAAGGCAAAAAGATCTTTGGCACCCAGA TTTGACCTTCCTGACATGAAAGAAACAAAGTATACTGTGGACAAGAGGTTTGGCATGGATTTTAAAGAAATAGAATTAATTG GCTCAGGTGGATTTGGCCAAGTTTTCAAAGCAAAACACAGAATTGACGGAAAGACTTACGTTATTAAACGTGTTAAATATAA TAACGAGAAGGCGGAGCGTGAAGTAAAAGCATTGGCAAAACTTGATCATGTAAATATTGTTCACTACAATGGCTGTTGGGAT GGATTTGATTATGATCCTGAGACCAGTGATGATTCTCTTGAGAGCAGTGATTATGATCCTGAGAACAGCAAAAATAGTTCAA GGTCAAAGACTAAGTGCCTTTTCATCCAAATGGAATTCTGTGATAAAGGGACCTTGGAACAATGGATTGAAAAAAGAAGAGG CGAGAAACTAGACAAAGTTTTGGCTTTGGAACTCTTTGAACAAATAACAAAAGGGGTGGATTATATACATTCAAAAAAATTA ATTCATAGAGATCTTAAGCCAAGTAATATATTCTTAGTAGATACAAAACAAGTAAAGATTGGAGACTTTGGACTTGTAACAT CTCTGAAAAATGATGGAAAGCGAACAAGGAGTAAGGGAACTTTGCGATACATGAGCCCAGAACAGATTTCTTCGCAAGACTA TGGAAAGGAAGTGGACCTCTACGCTTTGGGGCTAATTCTTGCTGAACTTCTTCATGTATGTGACACTGCTTTTGAAACATCA AAGTTTTTCACAGACCTACGGGATGGCATCATCTCAGATATATTTGATAAAAAAGAAAAAACTCTTCTACAGAAATTACTCT CAAAGAAACCTGAGGATCGACCTAACACATCTGAAATACTAAGGACCTTGACTGTGTGGAAGAAAAGCCCAGAGAAAAATGA ACGACACACATGTTAG HumanTTPCDS (SEQIDNO:48) ATGGCGGCTCAGCGGATCCGAGCGGCCAACTCCAATGGCCTCCCTCGCTGCAAGTCAGAGGGGACCCTGATTGACCTGAGCG AAGGGTTTTCAGAGACGAGCTTTAATGACATCAAAGTGCCTTCTCCCAGTGCCTTGCTCGTAGACAACCCCACACCTTTCGG AAATGCAAAGGAAGTGATTGCGATCAAGGACTATTGCCCCACCAACTTCACCACACTGAAGTTCTCCAAGGGCGACCATCTC TACGTCTTGGACACATCTGGCGGTGAGTGGTGGTACGCACACAACACCACCGAAATGGGCTACATCCCCTCCTCCTATGTGC AGCCCTTGAACTACCGGAACTCAACACTGAGTGACAGCGGTATGATTGATAATCTTCCAGACAGCCCAGACGAGGTAGCCAA GGAGCTGGAGCTGCTCGGGGGATGGACAGATGACAAAAAAGTACCAGGCAGAATGTACAGTAATAACCCTTTCTGGAATGGG GTCCAGACCAATCCATTTCTGAATGGGAACGTGCCCGTCATGCCCAGCCTGGATGAGCTGAATCCCAAAAGTACTGTGGATT TGCTCCTTTTTGACGCAGGTACATCCTCCTTCACCGAATCCAGCTCAGCCACCACGAATAGCACTGGCAACATCTTCGATGA GCTTCCAGTCACAAACGGACTCCACGCAGAGCCGCCGGTCAGGCGGGACAACCCCTTCTTCAGAAGCAAGCGCTCCTACAGT CTCTCGGAACTCTCCGTCCTCCAAGCCAAGTCCGATGCTCCCACATCGTCGAGTTTCTTCACCGGCTTGAAATCACCTGCCC CCGAGCAATTTCAGAGCCGGGAGGATTTTCGAACTGCCTGGCTAAACCACAGGAAGCTGGCCCGGTCTTGCCACGACCTGGA CTTGCTTGGCCAAAGCCCTGGTTGGGGCCAGACCCAAGCCGTGGAGACAAACATCGTGTGCAAGCTGGATAGCTCCGGGGGT GCTGTCCAGCTTCCTGACACCAGCATCAGCATCCACGTGCCCGAGGGCCACGTCGCCCCTGGGGAGACCCAGCAGATCTCCA TGAAAGCCCTGCTGGACCCCCCGCTGGAGCTCAACAGTGACAGGTCCTGCAGCATCAGCCCTGTGCTGGAGGTCAAGCTGAG CAACCTGGAGGTGAAAACCTCTATCATCTTGGAGATGAAAGTGTCAGCCGAGATAAAAAATGACCTTTTTAGCAAAAGCACA GTGGGCCTCCAGTGCCTGAGGAGCGACTCGAAGGAAGGGCCATATGTCTCCGTCCCGCTCAACTGCAGCTGTGGGGACACGG TCCAGGCACAGCTGCACAACCTGGAGCCCTGTATGTACGTGGCTGTCGTGGCCCATGGCCCAAGCATCCTCTACCCTTCCAC CGTGTGGGACTTCATCAATAAAAAAGTCACAGTGGGTCTCTACGGCCCTAAACACATCCACCCATCCTTCAAGACGGTAGTG ACCATTTTTGGGCATGACTGTGCCCCAAAGACGCTCCTGGTCAGCGAGGTCACACGCCAGGCACCCAACCCTGCCCCGGTGG CCCTGCAGCTGTGGGGGAAGCACCAGTTCGTTTTGTCCAGGCCCCAGGATCTCAAGGTCTGTATGTTTTCCAATATGACGAA TTACGAGGTCAAAGCCAGCGAGCAGGCCAAAGTGGTGCGAGGATTCCAGCTGAAGCTGGGCAAGGTGAGCCGCCTGATCTTC CCCATCACCTCCCAGAACCCCAACGAGCTCTCTGACTTCACGCTGCGGGTTCAGGTGAAGGACGACCAGGAGGCCATCCTCA CCCAGTTTTGTGTCCAGACTCCTCAGCCACCCCCTAAAAGTGCCATCAAGCCTTCCGGGCAAAGGAGGTTTCTCAAGAAGAA CGAAGTCGGGAAAATCATCCTGTCCCCGTTTGCCACCACTACAAAGTACCCGACTTTCCAGGACCGCCCGGTGTCCAGCCTC AAGTTTGGTAAGTTGCTCAAGACTGTGGTGCGGCAGAACAAGAACCACTACCTGCTGGAGTACAAGAAGGGCGACGGGATCG CCCTGCTCAGCGAGGAGCGGGTCAGGCTCCGGGGCCAGCTGTGGACCAAGGAGTGGTACATCGGCTACTACCAGGGCAGGGT GGGCCTCGTGCACACCAAGAACGTGCTGGTGGTCGGCAGGGCCCGGCCCAGCCTGTGCTCGGGCCCCGAGCTGAGCACCTCG GTGCTGCTGGAGCAGATCCTGCGGCCCTGCAAATTCCTCACGTACATCTATGCCTCCGTGAGGACCCTGCTCATGGAGAACA TCAGCAGCTGGCGCTCCTTCGCTGACGCCCTGGGCTACGTGAACCTGCCGCTCACCTTTTTCTGCCGGGCAGAGCTGGATAG TGAGCCCGAGCGGGTGGCGTCCGTCCTAGAAAAGCTGAAGGAGGACTGTAACAACACTGAGAACAAAGAACGGAAGTCCTTC CAGAAGGAGCTTGTGATGGCCCTACTGAAGATGGACTGCCAGGGCCTGGTGGTCAGACTCATCCAGGACTTTGTGCTCCTGA CCACGGCTGTAGAGGTGGCCCAGCGCTGGCGGGAGCTGGCTGAGAAGCTGGCCAAGGTCTCCAAGCAGCAGATGGACGCCTA CGAGTCTCCCCACCGGGACAGGAACGGGGTTGTGGACAGCGAGGCCATGTGGAAGCCTGCGTATGACTTCTTACTCACCTGG AGCCATCAGATCGGGGACAGCTACCGGGATGTCATCCAGGAGCTGCACCTGGGCCTGGACAAGATGAAAAACCCCATCACCA AGCGCTGGAAGCACCTCACTGGGACTCTGATCTTGGTGAACTCCCTGGACGTTCTGAGAGCAGCCGCCTTCAGCCCTGCGGA CCAGGACGACTTCGTGATTTGA HumanNEK2CDS (SEQIDNO:49) ATGCTGTCCAACTCCCAGGGCCAGAGCCCGCCGGTGCCGTTCCCCGCCCCGGCCCCGCCGCCGCAGCCCCCCACCCCTGCCC TGCCGCACCCCCCGGCGCAGCCGCCGCCGCCGCCCCCGCAGCAGTTCCCGCAGTTCCACGTCAAGTCCGGCCTGCAGATCAA GAAGAACGCCATCATCGATGACTACAAGGTCACCAGCCAGGTCCTGGGGCTGGGCATCAACGGCAAAGTTTTGCAGATCTTC AACAAGAGGACCCAGGAGAAATTCGCCCTCAAAATGCTTCAGGACTGCCCCAAGGCCCGCAGGGAGGTGGAGCTGCACTGGC GGGCCTCCCAGTGCCCGCACATCGTACGGATCGTGGATGTGTACGAGAATCTGTACGCAGGGAGGAAGTGCCTGCTGATTGT CATGGAATGTTTGGACGGTGGAGAACTCTTTAGCCGAATCCAGGATCGAGGAGACCAGGCATTCACAGAAAGAGAAGCATCC GAAATCATGAAGAGCATCGGTGAGGCCATCCAGTATCTGCATTCAATCAACATTGCCCATCGGGATGTCAAGCCTGAGAATC TCTTATACACCTCCAAAAGGCCCAACGCCATCCTGAAACTCACTGACTTTGGCTTTGCCAAGGAAACCACCAGCCACAACTC TTTGACCACTCCTTGTTATACACCGTACTATGTGGCTCCAGAAGTGCTGGGTCCAGAGAAGTATGACAAGTCCTGTGACATG TGGTCCCTGGGTGTCATCATGTACATCCTGCTGTGTGGGTATCCCCCCTTCTACTCCAACCACGGCCTTGCCATCTCTCCGG GCATGAAGACTCGCATCCGAATGGGCCAGTATGAATTTCCCAACCCAGAATGGTCAGAAGTATCAGAGGAAGTGAAGATGCT CATTCGGAATCTGCTGAAAACAGAGCCCACCCAGAGAATGACCATCACCGAGTTTATGAACCACCCTTGGATCATGCAATCA ACAAAGGTCCCTCAAACCCCACTGCACACCAGCCGGGTCCTGAAGGAGGACAAGGAGCGGTGGGAGGATGTCAAGGGGTGTC TTCATGACAAGAACAGCGACCAGGCCACTTGGCTGACCAGGTTGTGA

[0336] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0337] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-1<B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein described herein. Antisense nucleic acids targeting a nucleic acid encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein can be designed using the software available at the Integrated DNA Technologies website.

[0338] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0339] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0340] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, LBP, TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0341] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, -units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0342] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein (e.g., specificity for a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA, e.g., specificity for any one of SEQ ID NOs: 13-49). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can be designed based upon the nucleotide sequence of any of the TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0343] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0344] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Natl. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0345] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0346] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0347] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Natl. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res., 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0348] Another means by which expression of a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0349] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0350] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0351] The siRNA molecules used to decrease expression of a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0352] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, INK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 13-49, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0353] Exemplary TNF inhibitors that are inhibitory nucleic acids targeting TNF include, e.g., antisense DNA (e.g., Myers et al., J Pharmacol Exp Ther. 304(1):411-424, 2003; Wasmuth et al., Invest. Opthalmol. Vis. Sci, 2003; Dong et al., J. Orthop. Res. 26(8):1114-1120, 2008; U.S. Patent Application Serial Nos. 2003/0083275, 2003/0022848, and 2004/0770970; ISIS 104838; U.S. Pat. Nos. 6,180,403, 6,080,580, and 6,228,642; Kobzik et al., Inhibition of TNF Synthesis by Antisense Oligonucleotides, in Manual of Antisense Methodology, Kluwer Academic Publishers, Vol. 4, pp. 107-123, 1999; Taylor et al., Antisense Nucleic Acid Drug Develop. 8(3):199-205, 1998; Mayne et al., Stroke 32:240-248, 2001; Mochizuki et al., J. Controlled Release 151(2):155-161, 2011; Dong et al., J. Orthopaedic Res. 26(8):1114-1120, 2008; Dong et al., Pharm. Res. 28(6):1349-1356, 2011; and Pampfer et al., Biol. Reproduction 52 (6):1316-1326, 1995), antisense RNA, short interfering RNA (siRNA) (e.g., Taishi et al., Brain Research 1156:125-132, 2007; Presumey et al., Eur. J. Pharm. Biopharm. 82(3):457-467, 2012; Laroui et al., J. Controlled Release 186:41-53, 2014; D'Amore et al., Int. J. Immunopathology Pharmacol. 21:1045-1047, 2008; Choi et al., J. Dermatol. Sci. 52:87-97, 2008; Qin et al., Artificial Organs 35:706-714, 2011; McCarthy et al., J. Controlled Release 168: 28-34, 2013; Khoury et al., Current Opin. Mol. Therapeutics 9(5):483-489, 2007; Lu et al., RNA Interference Technology From Basic Science to Drug Development 303, 2005; Xie et al., PharmaGenomics 4(6):28-34, 2004; Aldawsari et al., Current Pharmaceutical Design 21(31):4594-4605, 2015; Zheng et al., Arch. Med. Sci. 11:1296-1302, 2015; Peng et al., Chinese J. Surgery 47(5):377-380, 2009; Aldayel et al., Molecular Therapy. Nucleic Acids 5(7):e340, 2016; Bai et al., Current Drug Targets 16:1531-1539, 2015; U.S. Patent Application Publications Nos. 2008/0097091, 2009/0306356, and 2005/0227935; and WO 14/168264), short hairpin RNA (shRNA) (e.g., Jakobsen et al., Mol. Ther. 17(10): 1743-1753, 2009; Ogawa et al., PLoS One 9(3): e92073, 2014; Ding et al., Bone Joint 94-6(Suppl. 11):44, 2014; and Hernandez-Alejandro et al., J. Surgical Res. 176(2):614-620, 2012), and microRNAs (see, e.g., WO 15/26249). In some embodiments, the inhibitory nucleic acid blocks pre-mRNA splicing of TNF (e.g., Chiu et al., Mol. Pharmacol. 71(6): 1640-1645, 2007).

[0354] In some embodiments, the inhibitory nucleic acid, e.g., an aptamer (e.g., Orava et al., ACS Chem Biol. 2013; 8(1): 170-178, 2013), can block the binding of a TNF protein with its receptor (TNFR1 and/or TNFR2).

[0355] In some embodiments, the inhibitory nucleic acid can down-regulate the expression of a TNF-induced downstream mediator (e.g., TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, p38, JNK, IB-, or CCL2). Further teachings of downstream TNF-induced mediators can be found in, e.g., Schwamborn et al., BMC Genomics 4:46, 2003; and Zhou et al., Oncogene 22: 2034-2044, 2003, incorporated by reference herein. Additional aspects of inhibitory nucleic acids are described in Aagaard et al., Adv. Drug Delivery Rev. 59(2):75-86, 2007, and Burnett et al., Biotechnol. J. 6(9):1130-1146, 2011.

[0356] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein can be administered to a subject (e.g., a human subject) in need thereof.

[0357] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0358] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0359] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0360] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of TNF, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-B, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IB, NF-B, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2) with a Tm of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive),

[0361] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0362] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0363] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0364] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0365] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0366] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0367] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0368] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0369] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0370] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0371] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0372] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0373] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0374] In some embodiments, the TNF inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of TNF, TNFR1, or TNFR2. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to TNF. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to an TNF receptor (TNFR1 or TNFR2).

[0375] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv)2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and seFv1-PEG-seFv2.

[0376] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab)2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0377] Non-limiting examples of TNF inhibitors that are antibodies that specifically bind to TNF are described in Elliott et al., Lancet 1994; 344: 1125-1127, 1994; Rankin et al., Br. J. Rheumatol. 2:334-342, 1995; Butler et al., Eur. Cytokine Network 6(4):225-230, 1994; Lorenz et al., J. Immunol. 156(4):1646-1653, 1996; Hinshaw et al., Circulatory Shock 30(3):279-292, 1990; Wanner et al., Shock 11(6):391-395, 1999; Bongartz et al., JAMA 295(19):2275-2285, 2006; Knight et al., Molecular Immunol. 30(16):1443-1453, 1993; Feldman, Nature Reviews Immunol. 2(5):364-371, 2002; Taylor et al., Nature Reviews Rheumatol. 5(10):578-582, 2009; Garces et al., Annals Rheumatic Dis. 72(12):1947-1955, 2013; Palladino et al., Nature Rev. Drug Discovery 2(9):736-746, 2003; Sandborn et al., Inflammatory Bowel Diseases 5(2):119-133, 1999; Atzeni et al., Autoimmunity Reviews 12(7):703-708, 2013; Maini et al., Immunol. Rev. 144(1):195-223, 1995; Ordas et al., Clin. Pharmacol. Therapeutics 91(4):635-646, 2012; Cohen et al., Canadian J Gastroenterol. Hepatol. 15(6):376-384, 2001; Feldmann et al., Ann. Rev. Immunol. 19(1):163-196, 2001; Ben-Horin et al., Autoimmunity Rev. 13(1):24-30, 2014; and U.S. Pat. Nos. 6,090,382; 6,258,562; and 6,509,015).

[0378] In certain embodiments, the TNF inhibitor can include or is infliximab (Remicade), CDP571, CDP 870, golimumab (Golimumab), adalimumab (Humira), or certolizumab pegol (Cimzia). In certain embodiments, the TNF inhibitor can be a TNF inhibitor biosimilar. Examples of approved and late-phase TNF inhibitor biosimilars include, but are not limited to, infliximab biosimilars such as Remsima and Inflectra (CT-P13) from Celltrion/Pfizer, GS071 from Aprogen, Flixabi (SB2) from Samsung Bioepis, PF-06438179 from Pfizer/Sandoz, NI-071 from Nichi-Iko Pharmaceutical Co., and ABP 710 from Amgen; adalimumab biosimilars such as Exemptia (ZRC3197) from Zydus Cadila, India, Solymbic and Amgevita (ABP 501) from Amgen, Imraldi (SB5) from Samsung Bioepis, GP-2017 from Sandoz, Switzerland, ONS-3010 from Oncobiologics/Viropro, U.S.A., M923 from Momenta Pharmaceuticals/Baxalta (Baxter spinoff USA), PF-06410293 from Pfizer, BMO-2 or MYL-1401-A from Biocon/Mylan, CHS-1420 from Coherus, FKB327 from Fujifilm/Kyowa Hakko Kirin (Fujifilm Kyowa Kirin Biologics), Cyltezo (BI 695501) from Boehringer Ingelheim, CT-P17 from Celltrion, BAX 923 from Baxalta (now a part of Shire), MSB11022 from Fresenius Kabi (bought from Merck kGaA (Merck Group) in 2017), LBAL from LG Life Sciences/Mochida Pharmaceutical, South Korea/Japan, PBP1502 from Prestige Biopharma, Adfrar from Torrent Pharmaceuticals, India, a biosimilar of adalimumab in development by Adello Biologics, a biosimilar of adalimumab in development by AET Biotech/BioXpress Therapeutics, Germany/Switzerland, a biosimilar of adalimumab from mAbxience, Spain, a biosimilar of adalimumab in development by PlantForm, Canada; and etanercept biosimilars such as Erelzi from Sandoz/Novartis, Brenzys (SB4) from Samsung Bioepis, GP2015 from Sandoz, TuNEX from Mycenax, LBEC0101 from LG Life, and CHS-0214 from Coherus.

[0379] In some embodiments, a biosimilar is an antibody or antigen-binding fragment thereof that has a light chain that has the same primary amino acid sequence as compared to a reference antibody (e.g., adalimumab) and a heavy chain that has the same primary amino acid sequence as compared to the reference antibody. In some examples, a biosimilar is an antibody or antigen-binding fragment thereof that has a light chain that includes the same light chain variable domain sequence as a reference antibody (e.g., adalimumab) and a heavy chain that includes the same heavy chain variable domain sequence as a reference antibody. In some embodiments, a biosimilar can have a similar glycosylation pattern as compared to the reference antibody (e.g., adalimumab). In other embodiments, a biosimilar can have a different glycosylation pattern as compared to the reference antibody (e.g., adalimumab).

[0380] Changes in the N-linked glycosylation profile of a biosimilar as compared to a reference antibody (e.g., adalimumab) can be detected using 2-anthranilic acid (AA)-derivatization and normal phase liquid chromatography with fluorescence detection, as generally described in Kamoda et al., J. Chromatography J. 1133:332-339, 2006. For example, a biosimilar can have changes in one or more (e.g., two, three, four, five, six, seven, eight, nine, ten, or eleven) of the following types of N-glycosylation as compared to the reference antibody (e.g., adalimumab): neutrally-charged oligosaccharides; monosialylated fucose-containing oligosaccharides; monosialylated oligosaccharides; bisialylated fucose-containing oligosaccharide; bisialylated oligosaccharides; triantennary, trisiaylated oligosaccharides of form 1; triantennary, trisialylated oligosaccharides of form 2; mannose-6-phosphate oligosaccharides; monophosphorylated oligosaccharides; tetrasialylated oligosaccharides; monosialylated and monophosphorylated oligosaccharides; and bis-mannose-6-phosphate oligosaccharides.

[0381] In some embodiments, the biosimilar can have a change in one, two, or three of: the percentage of species having one C-terminal lysine, the percentage of species having two C-terminal lysines, and the percentage of species having three C-terminal lysines as compared to the reference antibody (e.g., adalimumab).

[0382] In some embodiments, the biosimilar can have a change in the level of one, two, or three of acidic species, neutral species, and basic species in the composition as compared to the reference antibody (e.g., adalimumab).

[0383] In some embodiments, the biosimilar can have a change in the level of sulfation as compared to the reference antibody.

[0384] In some embodiments, the TNF inhibitor can be SAR252067 (e.g., a monoclonal antibody that specifically binds to TNFSF14, described in U.S. Patent Application Publication No. 2013/0315913) or MDGN-002 (described in U.S. Patent Application Publication No. 2015/0337046). In some embodiments, the TNF inhibitor can be PF-06480605, which binds specifically to TNFSF15 (e.g., described in U.S. Patent Application Publication No. 2015/0132311). Additional examples of TNF inhibitors include DLCX105 (described in Tsianakas et al., Exp. Dermatol. 25:428-433, 2016) and PF-06480605, which binds specifically to TNFSF15 (described in U.S. Patent Application Publication No. 2015/0132311). Further examples of TNF inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., WO 17/158097, EP 3219727, WO 16/156465, and WO 17/167997.

[0385] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5M (e.g., less than 0.510.sup.5M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7M, less than 0.510.sup.7 M, less than 110.sup.8M, less than 0.510.sup.8M, less than 110.sup.9M, less than 0.510.sup.9M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11M, less than 0.510.sup.11M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0386] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11M (inclusive); about 0.510.sup.11M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.6 M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11M (inclusive); about 110.sup.11M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9M, about 0.510.sup.9M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7M, about 110.sup.8M, about 0.510.sup.8M, about 110.sup.9 M, or about 0.510.sup.9M (inclusive); about 0.510.sup.9M to about 110.sup.5M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8M, or about 110.sup.9M (inclusive); about 110.sup.9M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8M, or about 0.510.sup.8M (inclusive); about 0.510.sup.8M to about 110.sup.5 M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8M (inclusive); about 110.sup.8M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7M (inclusive); about 0.510.sup.7 M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5M, about 0.510.sup.5M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5M, about 0.510.sup.5M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5M or about 0.510.sup.5M (inclusive); or about 0.510.sup.5M to about 110.sup.5M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0387] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0388] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1(inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1(inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1(inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins

[0389] In some embodiments, the TNF inhibitory agent is a fusion protein (e.g., an extracellular domain of a TNFR fused to a partner peptide, e.g., an Fc region of an immunoglobulin, e.g., human IgG) (see, e.g., Peppel et al., J. Exp. Med. 174(6):1483-1489, 1991; Deeg et al., Leukemia 16(2):162, 2002) or a soluble TNFR (e.g., TNFR1 or TNFR2) that binds specifically to TNF. In some embodiments, the TNF inhibitor includes or is etanercept (Enbrel) (see, e.g., WO 91/03553 and WO 09/406,476, incorporated by reference herein). In some embodiments, the TNF inhibitor includes or is r-TBP-I (e.g., Gradstein et al., J. Acquir. Immune Defic. Syndr. 26(2): 111-117, 2001). In some embodiments, the TNF inhibitor includes or is a soluble TNF receptor (e.g., Watt et al., J Leukoc Biol. 66(6):1005-1013, 1999; Tsao et al., Eur Respir J. 14(3):490-495, 1999; Kozak et al., Am. J. Physiol. Reg. Integrative Comparative Physiol. 269(1):R23-R29, 1995; Mohler et al., J. Immunol. 151(3):1548-1561, 1993; Nophar et al., EMBO J. 9(10):3269, 1990; Bjornberg et al., Lymphokine Cytokine Res. 13(3):203-211, 1994; Piguet et al., Eur. Respiratory J. 7(3):515-518, 1994; and Gray et al., Proc. Natl. Acad. Sci. U.S.A. 87(19):7380-7384, 1990).

Small Molecules

[0390] In some embodiments, the TNF inhibitor is a small molecule. In some embodiments, the TNF inhibitor is C87 (Ma et al., J. Biol. Chem. 289(18):12457-66, 2014). In some embodiments, the small molecule is LMP-420 (e.g., Haraguchi et al., AIDS Res. Ther. 3:8, 2006). In some embodiments, the small molecule is a tumor necrosis factor-converting enzyme (TACE) inhibitor (e.g., Moss et al., Nature Clinical Practice Rheumatology 4: 300-309, 2008). In some embodiments, the TACE inhibitor is TMI-005 and BMS-561392. Additional examples of small molecule inhibitors are described in, e.g., He et al., Science 310(5750):1022-1025, 2005.

[0391] In some examples, the TNF inhibitor is a small molecule that inhibits the activity of one of TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, and NF-B, in a mammalian cell.

[0392] In some examples, the TNF inhibitor is a small molecule that inhibits the activity of one of CD14, MyD88 (see, e.g., Olson et al., Scientific Reports 5:14246, 2015), IRAK (Chaudhary et al., J. Med. Chem. 58(1):96-110, 2015), lipopolysaccharide binding protein (LBP) (see, e.g., U.S. Pat. No. 5,705,398), TRAF6 (e.g., 3-[(2,5-Dimethylphenyl)amino]-1-phenyl-2-propen-1-one), ras (e.g., Baker et al., Nature 497:577-578, 2013), raf (e.g., vemurafenib (PLX4032, RG7204), sorafenib tosylate, PLX-4720, dabrafenib (GSK2118436), GDC-0879, RAF265 (CHIR-265), AZ 628, NVP-BHG712, SB590885, ZM 336372, sorafenib, GW5074, TAK-632, CEP-32496, encorafenib (LGX818), CCT196969, LY3009120, R05126766 (CH5126766), PLX7904, and MLN2480), MEK1/2 (e.g., Facciorusso et al., Expert Review Gastroentrol. Hepatol. 9:993-1003, 2015), ERK1/2 (e.g., Mandal et al., Oncogene 35:2547-2561, 2016), NIK (e.g., Mortier et al., Bioorg. Med. Chem. Lett. 20:4515-4520, 2010), IKK (e.g., Reilly et al., Nature Med. 19:313-321, 2013), IB (e.g., Suzuki et al., Expert. Opin. Invest. Drugs 20:395-405, 2011), NF-B (e.g., Gupta et al., Biochim. Biophys. Acta 1799(10-12):775-787, 2010), rac (e.g., U.S. Pat. No. 9,278,956), MEK4/7, JNK (e.g., AEG 3482, BI 78D3, CEP 1347, c-JUN peptide, IQ 1S, JIP-1 (153-163), SP600125, SU 3327, and TCS JNK6o), c-jun (e.g., AEG 3482, BI 78D3, CEP 1347, c-JUN peptide, IQ 1S, JIP-1 (153-163), SP600125, SU 3327, and TCS JNK6o), MEK3/6 (e.g., Akinleye et al., J. Hematol. Oncol. 6:27, 2013), p38 (e.g., AL 8697, AMG 548, BIRB 796, CMPD-1, DBM 1285 dihydrochloride, EO 1428, JX 401, ML 3403, Org 48762-0, PH 797804, RWJ 67657, SB 202190, SB 203580, SB 239063, SB 706504, SCIO 469, SKF 86002, SX 011, TA 01, TA 02, TAK 715, VX 702, and VX 745), PKR (e.g., 2-aminopurine or CAS 608512-97-6), TTP (e.g., CAS 329907-28-0), and MK2 (PF 3644022 and PHA 767491).

IL-6 Receptor Inhibitors

[0393] The term IL-6 receptor inhibitor refers to an agent which decreases IL-6 receptor expression and/or the ability of IL-6 to bind to an IL-6 receptor. In some embodiments, the IL-6 receptor inhibitor targets the IL-6 receptor -subunit, glycoprotein 130 (sIL6gp130). In other embodiments, the IL-6 receptor inhibitor targets the IL-6 receptor subunit (IL6R). In other embodiments, the IL-6 receptor inhibitor targets the complex consisting of both the IL-6 receptor subunit (IL6R) and the IL-6 receptor -subunit, glycoprotein 130 (sIL6gp130). In some embodiments, the IL-6 receptor inhibitor targets IL-6.

[0394] In some embodiments, an IL-6 receptor inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, a IL-6 receptor antagonist, or a small molecule. In some embodiments, the inhibitory nucleic acid is a small interfering RNA, an antisense nucleic acid, an aptamer, or a microRNA. Exemplary IL-6 receptor inhibitors are described herein. Additional examples of IL-6 receptor inhibitors are known in the art.

[0395] Exemplary aspects of different inhibitory nucleic acids are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of an IL6R, sIL6gp130, or IL-6 mRNA. Inhibitory nucleic acids that can decrease the expression of IL6R, sIL6gp130, or IL-6 mRNA in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL6R, sIL6gp130, or IL-6 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 50-55).

TABLE-US-00003 HumanIL6RmRNAVariant1 (SEQIDNO:50) 1 ggcggtcccctgttctccccgctcaggtgcggcgctgtggcaggaagccaccccctcggt 61 cggccggtgcgcggggctgttgcgccatccgctccggctttcgtaaccgcaccctgggac 121 ggcccagagacgctccagcgcgagttcctcaaatgttttcctgcgttgccaggaccgtcc 181 gccgctctgagtcatgtgcgagtgggaagtcgcactgacactgagccgggccagagggag 241 aggagccgagcgcggcgcggggccgagggactcgcagtgtgtgtagagagccgggctcct 301 gcggatgggggctgcccccggggcctgagcccgcctgcccgcccaccgccccgccccgcc 361 cctgccacccctgccgcccggttcccattagcctgtccgcctctgcgggaccatggagtg 421 gtagccgaggaggaagcatgctggccgtcggctgcgcgctgctggctgccctgctggccg 481 cgccgggagcggcgctggccccaaggcgctgccctgcgcaggaggtggcgagaggcgtgc 541 tgaccagtctgccaggagacagcgtgactctgacctgcccgggggtagagccggaagaca 601 atgccactgttcactgggtgctcaggaagccggctgcaggctcccaccccagcagatggg 661 ctggcatgggaaggaggctgctgctgaggtcggtgcagctccacgactctggaaactatt 721 catgctaccgggccggccgcccagctgggactgtgcacttgctggtggatgttccccccg 781 aggagccccagctctcctgcttccggaagagccccctcagcaatgttgtttgtgagtggg 841 gtcctcggagcaccccatccctgacgacaaaggctgtgctcttggtgaggaagtttcaga 901 acagtccggccgaagacttccaggagccgtgccagtattcccaggagtcccagaagttct 961 cctgccagttagcagtcccggagggagacagctctttctacatagtgtccatgtgcgtcg 1021 ccagtagtgtcgggagcaagttcagcaaaactcaaacctttcagggttgtggaatcttgc 1081 agcctgatccgcctgccaacatcacagtcactgccgtggccagaaacccccgctggctca 1141 gtgtcacctggcaagacccccactcctggaactcatctttctacagactacggtttgagc 1201 tcagatatcgggctgaacggtcaaagacattcacaacatggatggtcaaggacctccagc 1261 atcactgtgtcatccacgacgcctggagcggcctgaggcacgtggtgcagcttcgtgccc 1321 aggaggagttcgggcaaggcgagtggagcgagtggagcccggaggccatgggcacgcctt 1381 ggacagaatccaggagtcctccagctgagaacgaggtgtccacccccatgcaggcactta 1441 ctactaataaagacgatgataatattctcttcagagattctgcaaatgcgacaagcctcc 1501 cagtgcaagattcttcttcagtaccactgcccacattcctggttgctggagggagcctgg 1561 ccttcggaacgctcctctgcattgccattgttctgaggttcaagaagacgtggaagctgc 1621 gggctctgaaggaaggcaagacaagcatgcatccgccgtactctttggggcagctggtcc 1681 cggagaggcctcgacccaccccagtgcttgttcctctcatctccccaccggtgtccccca 1741 gcagcctggggtctgacaatacctcgagccacaaccgaccagatgccagggacccacgga 1801 gcccttatgacatcagcaatacagactacttcttccccagatagctggctgggtggcacc 1861 agcagcctggaccctgtggatgataaaacacaaacgggctcagcaaaagatgcttctcac 1921 tgccatgccagcttatctcaggggtgtgcggcctttggcttcacggaagagccttgcgga 1981 aggttctacgccaggggaaaatcagcctgctccagctgttcagctggttgaggtttcaaa 2041 cctccctttccaaatgcccagcttaaaggggctagagtgaacttgggccactgtgaagag 2101 aaccatatcaagactctttggacactcacacggacactcaaaagctgggcaggttggtgg 2161 gggcctcggtgtggagaagcggctggcagcccacccctcaacacctctgcacaagctgca 2221 ccctcaggcaggtgggatggatttccagccaaagcctcctccagccgccatgctcctggc 2281 ccactgcatcgtttcatcttccaactcaaactcttaaaacccaagtgccttagcaaattc 2341 tgtttttctaggcctggggacggcttttacttaaaccgccaaggctgggggaagaagctc 2401 tctcctccctttcttccctacagttgaaaaacagctgagggtgagtgggtgaataataca 2461 gtatctcagggcctggtcgttttcaacagaattataattagttcctcattagcattttgc 2521 taaatgtgaatgatgatcctaggcatttgctgaatacagaggcaactgcattggctttgg 2581 gttgcaggacctcaggtgagaagcagaggaaggagaggagaggggcacagggtctctacc 2641 atcccctgtagagtgggagctgagtgggggatcacagcctctgaaaaccaatgttctctc 2701 ttctccacctcccacaaaggagagctagcagcagggagggcttctgccatttctgagatc 2761 aaaacggttttactgcagctttgtttgttgtcagctgaacctgggtaactagggaagata 2821 atattaaggaagacaatgtgaaaagaaaaatgagcctggcaagaatgtgtttaaacttgg 2881 tttttaaaaaactgctgactgttttctcttgagagggtggaatatccaatattcgctgtg 2941 tcagcatagaagtaacttacttaggtgtgggggaagcaccataactttgtttagcccaaa 3001 accaagtcaagtgaaaaaggaggaagagaaaaaatattttcctgccaggcatggtggccc 3061 acgcacttcgggaggtcgaggcaggaggatcacttgagtccagaagtttgagatcagcct 3121 gggcaatgtgataaaaccccatctctacaaaaagcataaaaattagccaagtgtggtaga 3181 gtgtgcctgaagtcccagatacttggggggctgaggtgggaggatctcttgagcctggga 3241 ggtcaaggctgcagtgagccgagattgcaccactgcactccagcctgggtgacagagcaa 3301 gtgagaccctgtctcaaaaaaagaaaaagaaaaagaaaaaatattttccctattagagaa 3361 gagattgtggtttcattctgtattttgtttttgtcttaaaaagtggaaaaatagcctgcc 3421 tcttctctactctagggaaaaaccagcgtgtgactactcccccaggtggttatggagagg 3481 gtgtccggtccctgtcccagtgccgagaaggaagcctcccacgactgcccggcagggtcc 3541 tagaaattccccaccctgaaagccctgagctttctgctatcaaagaggttttaaaaaaat 3601 cccatttaaaaaaaatcccttacctcggtgccttcctctttttatttagttccttgagtt 3661 gattcagctctgcaagaattgaagcaggactaaatgtctagttgtaacaccatgattaac 3721 cacttcagctgacttttctgtccgagctttgaaaattcagtggtgttagtggttacccag 3781 ttagctctcaagttatcagggtattccagagtggggatatgatttaaatcagccgtgtaa 3841 ccatggacccaatatttaccagaccacaaaacttttctaatactctaccctcttagaaaa 3901 accaccaccatcaccagacaggtgcgaaaggatgaaagtgaccatgttttgtttacggtt 3961 ttccaggtttaagctgttactgtcttcagtaagccgtgattttcattgctgggcttgtct 4021 gtagattttagaccctattgctgcttgaggcaactcatcttaggttggcaaaaaggcagg 4081 atggccgggcgcggtggctcacgcctgtaatcctagcactttgggaggccaaggtgggag 4141 gattgcttgagctcaggagtttgagaccaacctgggtaacatagtgagacaccatctcta 4201 ttatgaacaataacagttaagaaaaaaaaaggcaggcaggcggttatggtggttccctcc 4261 catcccaccacataaagtttctgagacttgagaacagcaaaatgctgttaaagggaaata 4321 ttaagaatgagaatctgcagtaagggtgattctgtgcccacagttcttcaattctttata 4381 ccgttttacccacatgtggtgttaccaaagccgggcagaaccatgctagcggaagatgtg 4441 aaatccagatagctcattattgccaagagctaggcagctttgatctccaaattgttattg 4501 ctttcatttttattgtaatggaattgctttgttttgtttttttgtttttgtattgaagag 4561 ggttgttttccctttatttttcataagctaatgtaaatgaagaaaaaatgtcttctctgg 4621 gctgtaggcctggctcagcgtacacaggtatacatcctaagctctctatgttctctaatc 4681 tgtggtgactgaacatgtgtctcaatgcacggggcatttctacctgtgtttctgcagcac 4741 ccccactgccttgagtccccagcagtgctgttatttgcctaacacctgtagccatctgcc 4801 acgcagccagacgtgaaacgctgagacagagaccatttaggttaaatacgacagcttatc 4861 ctgctgggtggggaaagtaaaaaatatgctggttcaaggcctaaagtaaaatgatcaata 4921 atgtttgtagcattaatgaaatattttcaagaaatgtgtccaggggtagcactggctatg 4981 ttgacgaggcctttggtaactcagagagctcttggccctgatggggacttgcccttacgc 5041 tttctttatcaggctctgagttcacacggagcctctggcacttccctgctgtcttgggag 5101 aaaggaaactggttgccgcggcaggttgtggaatctgttgctggaaccaggctggaagcc 5161 cacctggtagtgaacagggcccagtggggcaggctgggcatgttgtggtctatgggtttg 5221 tttcctggagaatgttcaggaatgtcttcccagctgctttggtgctgagctctattatct 5281 cacagcacgtccagaaggctaacccaggtggggaggatgctgacaccagctccaggtgga 5341 gttggtggtcttaatttggagatgcaggggcaacctgtgaccctttgaggcaagagccct 5401 gcacccagctgtcccgtgcagccgtgggcaggggctgcacacggaggggcaggcgggcca 5461 gttcagggtccgtgccaggccctcctcagtgccctgtgaaggcctcctgtcctccgtgcg 5521 gctgggcaccagcaccagggagtttctatggcaaccttagtgattattaaggaacactgt 5581 cagttttatgaacatatgctcaaatgaaattctactttaggaggaaaggattggaacagc 5641 atgtcacaaggctgttaattaacagagagaccttattggatggagatcacatctgttaaa 5701 tagaatacctcaactctacgttgttttcttggagataaataatagtttcaagtttttgtt 5761 tgtttgttttacctaattacctgaaagcaaataccaaaggctgatgtctgtatatggggc 5821 aaagggtcagtatatttttcagtgtttttttttctaccagctattttgcatttaaagtga 5881 acattgtgtttggaataaatactcttaaaaaataaaaaaaaaaaaaaa HumanIL6RmRNAVariant2 (SEQIDNO:51) 1 ggcggtcccctgttctccccgctcaggtgcggcgctgtggcaggaagccaccccctcggt 61 cggccggtgcgcggggctgttgcgccatccgctccggctttcgtaaccgcaccctgggac 121 ggcccagagacgctccagcgcgagttcctcaaatgttttcctgcgttgccaggaccgtcc 181 gccgctctgagtcatgtgcgagtgggaagtcgcactgacactgagccgggccagagggag 241 aggagccgagcgcggcgcggggccgagggactcgcagtgtgtgtagagagccgggctcct 301 gcggatgggggctgcccccggggcctgagcccgcctgcccgcccaccgccccgccccgcc 361 cctgccacccctgccgcccggttcccattagcctgtccgcctctgcgggaccatggagtg 421 gtagccgaggaggaagcatgctggccgtcggctgcgcgctgctggctgccctgctggccg 481 cgccgggagcggcgctggccccaaggcgctgccctgcgcaggaggtggcgagaggcgtgc 541 tgaccagtctgccaggagacagcgtgactctgacctgcccgggggtagagccggaagaca 601 atgccactgttcactgggtgctcaggaagccggctgcaggctcccaccccagcagatggg 661 ctggcatgggaaggaggctgctgctgaggtcggtgcagctccacgactctggaaactatt 721 catgctaccgggccggccgcccagctgggactgtgcacttgctggtggatgttccccccg 781 aggagccccagctctcctgcttccggaagagccccctcagcaatgttgtttgtgagtggg 841 gtcctcggagcaccccatccctgacgacaaaggctgtgctcttggtgaggaagtttcaga 901 acagtccggccgaagacttccaggagccgtgccagtattcccaggagtcccagaagttct 961 cctgccagttagcagtcccggagggagacagctctttctacatagtgtccatgtgcgtcg 1021 ccagtagtgtcgggagcaagttcagcaaaactcaaacctttcagggttgtggaatcttgc 1081 agcctgatccgcctgccaacatcacagtcactgccgtggccagaaacccccgctggctca 1141 gtgtcacctggcaagacccccactcctggaactcatctttctacagactacggtttgagc 1201 tcagatatcgggctgaacggtcaaagacattcacaacatggatggtcaaggacctccagc 1261 atcactgtgtcatccacgacgcctggagcggcctgaggcacgtggtgcagcttcgtgccc 1321 aggaggagttcgggcaaggcgagtggagcgagtggagcccggaggccatgggcacgcctt 1381 ggacagaatccaggagtcctccagctgagaacgaggtgtccacccccatgcaggcactta 1441 ctactaataaagacgatgataatattctcttcagagattctgcaaatgcgacaagcctcc 1501 caggttcaagaagacgtggaagctgcgggctctgaaggaaggcaagacaagcatgcatcc 1561 gccgtactctttggggcagctggtcccggagaggcctcgacccaccccagtgcttgttcc 1621 tctcatctccccaccggtgtcccccagcagcctggggtctgacaatacctcgagccacaa 1681 ccgaccagatgccagggacccacggagcccttatgacatcagcaatacagactacttctt 1741 ccccagatagctggctgggtggcaccagcagcctggaccctgtggatgataaaacacaaa 1801 cgggctcagcaaaagatgcttctcactgccatgccagcttatctcaggggtgtgcggcct 1861 ttggcttcacggaagagccttgcggaaggttctacgccaggggaaaatcagcctgctcca 1921 gctgttcagctggttgaggtttcaaacctccctttccaaatgcccagcttaaaggggcta 1981 gagtgaacttgggccactgtgaagagaaccatatcaagactctttggacactcacacgga 2041 cactcaaaagctgggcaggttggtgggggcctcggtgtggagaagcggctggcagcccac 2101 ccctcaacacctctgcacaagctgcaccctcaggcaggtgggatggatttccagccaaag 2161 cctcctccagccgccatgctcctggcccactgcatcgtttcatcttccaactcaaactct 2221 taaaacccaagtgccttagcaaattctgtttttctaggcctggggacggcttttacttaa 2281 accgccaaggctgggggaagaagctctctcctccctttcttccctacagttgaaaaacag 2341 ctgagggtgagtgggtgaataatacagtatctcagggcctggtcgttttcaacagaatta 2401 taattagttcctcattagcattttgctaaatgtgaatgatgatcctaggcatttgctgaa 2461 tacagaggcaactgcattggctttgggttgcaggacctcaggtgagaagcagaggaagga 2521 gaggagaggggcacagggtctctaccatcccctgtagagtgggagctgagtgggggatca 2581 cagcctctgaaaaccaatgttctctcttctccacctcccacaaaggagagctagcagcag 2641 ggagggcttctgccatttctgagatcaaaacggttttactgcagctttgtttgttgtcag 2701 ctgaacctgggtaactagggaagataatattaaggaagacaatgtgaaaagaaaaatgag 2761 cctggcaagaatgtgtttaaacttggtttttaaaaaactgctgactgttttctcttgaga 2821 gggtggaatatccaatattcgctgtgtcagcatagaagtaacttacttaggtgtggggga 2881 agcaccataactttgtttagcccaaaaccaagtcaagtgaaaaaggaggaagagaaaaaa 2941 tattttcctgccaggcatggtggcccacgcacttcgggaggtcgaggcaggaggatcact 3001 tgagtccagaagtttgagatcagcctgggcaatgtgataaaaccccatctctacaaaaag 3061 cataaaaattagccaagtgtggtagagtgtgcctgaagtcccagatacttggggggctga 3121 ggtgggaggatctcttgagcctgggaggtcaaggctgcagtgagccgagattgcaccact 3181 gcactccagcctgggtgacagagcaagtgagaccctgtctcaaaaaaagaaaaagaaaaa 3241 gaaaaaatattttccctattagagaagagattgtggtttcattctgtattttgtttttgt 3301 cttaaaaagtggaaaaatagcctgcctcttctctactctagggaaaaaccagcgtgtgac 3361 tactcccccaggtggttatggagagggtgtccggtccctgtcccagtgccgagaaggaag 3421 cctcccacgactgcccggcagggtcctagaaattccccaccctgaaagccctgagctttc 3481 tgctatcaaagaggttttaaaaaaatcccatttaaaaaaaatcccttacctcggtgcctt 3541 cctctttttatttagttccttgagttgattcagctctgcaagaattgaagcaggactaaa 3601 tgtctagttgtaacaccatgattaaccacttcagctgacttttctgtccgagctttgaaa 3661 attcagtggtgttagtggttacccagttagctctcaagttatcagggtattccagagtgg 3721 ggatatgatttaaatcagccgtgtaaccatggacccaatatttaccagaccacaaaactt 3781 ttctaatactctaccctcttagaaaaaccaccaccatcaccagacaggtgcgaaaggatg 3841 aaagtgaccatgttttgtttacggttttccaggtttaagctgttactgtcttcagtaagc 3901 cgtgattttcattgctgggcttgtctgtagattttagaccctattgctgcttgaggcaac 3961 tcatcttaggttggcaaaaaggcaggatggccgggcgcggtggctcacgcctgtaatcct 4021 agcactttgggaggccaaggtgggaggattgcttgagctcaggagtttgagaccaacctg 4081 ggtaacatagtgagacaccatctctattatgaacaataacagttaagaaaaaaaaaggca 4141 ggcaggcggttatggtggttccctcccatcccaccacataaagtttctgagacttgagaa 4201 cagcaaaatgctgttaaagggaaatattaagaatgagaatctgcagtaagggtgattctg 4261 tgcccacagttcttcaattctttataccgttttacccacatgtggtgttaccaaagccgg 4321 gcagaaccatgctagcggaagatgtgaaatccagatagctcattattgccaagagctagg 4381 cagctttgatctccaaattgttattgctttcatttttattgtaatggaattgctttgttt 4441 tgtttttttgtttttgtattgaagagggttgttttccctttatttttcataagctaatgt 4501 aaatgaagaaaaaatgtcttctctgggctgtaggcctggctcagcgtacacaggtataca 4561 tcctaagctctctatgttctctaatctgtggtgactgaacatgtgtctcaatgcacgggg 4621 catttctacctgtgtttctgcagcacccccactgccttgagtccccagcagtgctgttat 4681 ttgcctaacacctgtagccatctgccacgcagccagacgtgaaacgctgagacagagacc 4741 atttaggttaaatacgacagcttatcctgctgggtggggaaagtaaaaaatatgctggtt 4801 caaggcctaaagtaaaatgatcaataatgtttgtagcattaatgaaatattttcaagaaa 4861 tgtgtccaggggtagcactggctatgttgacgaggcctttggtaactcagagagctcttg 4921 gccctgatggggacttgcccttacgctttctttatcaggctctgagttcacacggagcct 4981 ctggcacttccctgctgtcttgggagaaaggaaactggttgccgcggcaggttgtggaat 5041 ctgttgctggaaccaggctggaagcccacctggtagtgaacagggcccagtggggcaggc 5101 tgggcatgttgtggtctatgggtttgtttcctggagaatgttcaggaatgtcttcccagc 5161 tgctttggtgctgagctctattatctcacagcacgtccagaaggctaacccaggtgggga 5221 ggatgctgacaccagctccaggtggagttggtggtcttaatttggagatgcaggggcaac 5281 ctgtgaccctttgaggcaagagccctgcacccagctgtcccgtgcagccgtgggcagggg 5341 ctgcacacggaggggcaggcgggccagttcagggtccgtgccaggccctcctcagtgccc 5401 tgtgaaggcctcctgtcctccgtgcggctgggcaccagcaccagggagtttctatggcaa 5461 ccttagtgattattaaggaacactgtcagttttatgaacatatgctcaaatgaaattcta 5521 ctttaggaggaaaggattggaacagcatgtcacaaggctgttaattaacagagagacctt 5581 attggatggagatcacatctgttaaatagaatacctcaactctacgttgttttcttggag 5641 ataaataatagtttcaagtttttgtttgtttgttttacctaattacctgaaagcaaatac 5701 caaaggctgatgtctgtatatggggcaaagggtcagtatatttttcagtgtttttttttc 5761 taccagctattttgcatttaaagtgaacattgtgtttggaataaatactcttaaaaaata 5821 aaaaaaaaaaaaaa HumanIL6RmRNAVariant3 (SEQIDNO:52) 1 ggcggtcccctgttctccccgctcaggtgcggcgctgtggcaggaagccaccccctcggt 61 cggccggtgcgcggggctgttgcgccatccgctccggctttcgtaaccgcaccctgggac 121 ggcccagagacgctccagcgcgagttcctcaaatgttttcctgcgttgccaggaccgtcc 181 gccgctctgagtcatgtgcgagtgggaagtcgcactgacactgagccgggccagagggag 241 aggagccgagcgcggcgcggggccgagggactcgcagtgtgtgtagagagccgggctcct 301 gcggatgggggctgcccccggggcctgagcccgcctgcccgcccaccgccccgccccgcc 361 cctgccacccctgccgcccggttcccattagcctgtccgcctctgcgggaccatggagtg 421 gtagccgaggaggaagcatgctggccgtcggctgcgcgctgctggctgccctgctggccg 481 cgccgggagcggcgctggccccaaggcgctgccctgcgcaggaggtggcgagaggcgtgc 541 tgaccagtctgccaggagacagcgtgactctgacctgcccgggggtagagccggaagaca 601 atgccactgttcactgggtgctcaggaagccggctgcaggctcccaccccagcagatggg 661 ctggcatgggaaggaggctgctgctgaggtcggtgcagctccacgactctggaaactatt 721 catgctaccgggccggccgcccagctgggactgtgcacttgctggtggatgttccccccg 781 aggagccccagctctcctgcttccggaagagccccctcagcaatgttgtttgtgagtggg 841 gtcctcggagcaccccatccctgacgacaaaggctgtgctcttggtgaggaagtttcaga 901 acagtccggccgaagacttccaggagccgtgccagtattcccaggagtcccagaagttct 961 cctgccagttagcagtcccggagggagacagctctttctacatagtgtccatgtgcgtcg 1021 ccagtagtgtcgggagcaagttcagcaaaactcaaacctttcagggttgtggaatcttgc 1081 agcctgatccgcctgccaacatcacagtcactgccgtggccagaaacccccgctggctca 1141 gtgtcacctggcaagacccccactcctggaactcatctttctacagactacggtttgagc 1201 tcagatatcgggctgaacggtcaaagacattcacaacatggatggtcaaggacctccagc 1261 atcactgtgtcatccacgacgcctggagcggcctgaggcacgtggtgcagcttcgtgccc 1321 aggaggagttcgggcaaggcgagtggagcgagtggagcccggaggccatgggcacgcctt 1381 ggacagacaggctttctcctcgttgcccaggatggagtacagcagtgcaatcacagctca 1441 cggcaacttctgcctcctgggttcaagcaatcctcccgcctcagcctcctaagtagctgg 1501 gaccacaggcgtgtgccacaatgctaattttttaaaaatgttttgtagagacagggtttc 1561 accatgctgcccaggctggtctcgaactcctggcctcaagtgatccaccagcctcagact 1621 cccaaagtgctgggattactggtgtgagccactgcacctgactaaactttaaattttttt 1681 ttttagacggaatctcgctctgttgcccaggctggagtgcagtggcatgatattggctca 1741 ctgcaagctctgcctcttgggttcacgctattctcctgcctcagcctcctgagtagctgg 1801 gactacaggtgcacaccaccacgcccggctaatttttttttttttttagtagagacgggg 1861 tttcactgtgttggccaggctggtcttgaactcctgacctcgtgatccacccgcctcgcc 1921 ctcccaaaatgctgggattacaggtgtgagccaccgcgcctggcctaaacttttaaaatt 1981 ttaatcaaattaatacatgcacatggcaaagaagtaataaacagcttataacactgaaaa 2041 aaaaaaaaaaaaaaaaaa HumanIL-6receptor-subunit,glycoprotein130(sIL6gp130) (SEQIDNO:53) 1 gagcagccaaaaggcccgcggagtcgcgctgggccgccccggcgcagctgaaccgggggc 61 cgcgcctgccaggccgacgggtctggcccagcctggcgccaaggggttcgtgcgctgtgg 121 agacgcggagggtcgaggcggcgcggcctgagtgaaacccaatggaaaaagcatgacatt 181 tagaagtagaagacttagcttcaaatccctactccttcacttactaattttgtgatttgg 241 aaatatccgcgcaagatgttgacgttgcagacttgggtagtgcaagccttgtttattttc 301 ctcaccactgaatctacaggtgaacttctagatccatgtggttatatcagtcctgaatct 361 ccagttgtacaacttcattctaatttcactgcagtttgtgtgctaaaggaaaaatgtatg 421 gattattttcatgtaaatgctaattacattgtctggaaaacaaaccattttactattcct 481 aaggagcaatatactatcataaacagaacagcatccagtgtcacctttacagatatagct 541 tcattaaatattcagctcacttgcaacattcttacattcggacagcttgaacagaatgtt 601 tatggaatcacaataatttcaggcttgcctccagaaaaacctaaaaatttgagttgcatt 661 gtgaacgaggggaagaaaatgaggtgtgagtgggatggtggaagggaaacacacttggag 721 acaaacttcactttaaaatctgaatgggcaacacacaagtttgctgattgcaaagcaaaa 781 cgtgacacccccacctcatgcactgttgattattctactgtgtattttgtcaacattgaa 841 gtctgggtagaagcagagaatgcccttgggaaggttacatcagatcatatcaattttgat 901 cctgtatataaagtgaagcccaatccgccacataatttatcagtgatcaactcagaggaa 961 ctgtctagtatcttaaaattgacatggaccaacccaagtattaagagtgttataatacta 1021 aaatataacattcaatataggaccaaagatgcctcaacttggagccagattcctcctgaa 1081 gacacagcatccacccgatcttcattcactgtccaagaccttaaaccttttacagaatat 1141 gtgtttaggattcgctgtatgaaggaagatggtaagggatactggagtgactggagtgaa 1201 gaagcaagtgggatcacctatgaagatagaccatctaaagcaccaagtttctggtataaa 1261 atagatccatcccatactcaaggctacagaactgtacaactcgtgtggaagacattgcct 1321 ccttttgaagccaatggaaaaatcttggattatgaagtgactctcacaagatggaaatca 1381 catttacaaaattacacagttaatgccacaaaactgacagtaaatctcacaaatgatcgc 1441 tatctagcaaccctaacagtaagaaatcttgttggcaaatcagatgcagctgttttaact 1501 atccctgcctgtgactttcaagctactcaccctgtaatggatcttaaagcattccccaaa 1561 gataacatgctttgggtggaatggactactccaagggaatctgtaaagaaatatatactt 1621 gagtggtgtgtgttatcagataaagcaccctgtatcacagactggcaacaagaagatggt 1681 accgtgcatcgcacctatttaagagggaacttagcagagagcaaatgctatttgataaca 1741 gttactccagtatatgctgatggaccaggaagccctgaatccataaaggcataccttaaa 1801 caagctccaccttccaaaggacctactgttcggacaaaaaaagtagggaaaaacgaagct 1861 gtcttagagtgggaccaacttcctgttgatgttcagaatggatttatcagaaattatact 1921 atattttatagaaccatcattggaaatgaaactgctgtgaatgtggattcttcccacaca 1981 gaatatacattgtcctctttgactagtgacacattgtacatggtacgaatggcagcatac 2041 acagatgaaggtgggaaggatggtccagaattcacttttactaccccaaagtttgctcaa 2101 ggagaaattgaagccatagtcgtgcctgtttgcttagcattcctattgacaactcttctg 2161 ggagtgctgttctgctttaataagcgagacctaattaaaaaacacatctggcctaatgtt 2221 ccagatccttcaaagagtcatattgcccagtggtcacctcacactcctccaaggcacaat 2281 tttaattcaaaagatcaaatgtattcagatggcaatttcactgatgtaagtgttgtggaa 2341 atagaagcaaatgacaaaaagccttttccagaagatctgaaatcattggacctgttcaaa 2401 aaggaaaaaattaatactgaaggacacagcagtggtattggggggtcttcatgcatgtca 2461 tcttctaggccaagcatttctagcagtgatgaaaatgaatcttcacaaaacacttcgagc 2521 actgtccagtattctaccgtggtacacagtggctacagacaccaagttccgtcagtccaa 2581 gtcttctcaagatccgagtctacccagcccttgttagattcagaggagcggccagaagat 2641 ctacaattagtagatcatgtagatggcggtgatggtattttgcccaggcaacagtacttc 2701 aaacagaactgcagtcagcatgaatccagtccagatatttcacattttgaaaggtcaaag 2761 caagtttcatcagtcaatgaggaagattttgttagacttaaacagcagatttcagatcat 2821 atttcacaatcctgtggatctgggcaaatgaaaatgtttcaggaagtttctgcagcagat 2881 gcttttggtccaggtactgagggacaagtagaaagatttgaaacagttggcatggaggct 2941 gcgactgatgaaggcatgcctaaaagttacttaccacagactgtacggcaaggcggctac 3001 atgcctcagtgaaggactagtagttcctgctacaacttcagcagtacctataaagtaaag 3061 ctaaaatgattttatctgtgaattc HumanIL-6mRNATranscript1 (SEQIDNO:54) 1 gtctcaatattagagtctcaacccccaataaatataggactggagatgtctgaggctcat 61 tctgccctcgagcccaccgggaacgaaagagaagctctatctcccctccaggagcccagc 121 tatgaactccttctccacaagcgccttcggtccagttgccttctccctggggctgctcct 181 ggtgttgcctgctgccttccctgccccagtacccccaggagaagattccaaagatgtagc 241 cgccccacacagacagccactcacctcttcagaacgaattgacaaacaaattcggtacat 301 cctcgacggcatctcagccctgagaaaggagacatgtaacaagagtaacatgtgtgaaag 361 cagcaaagaggcactggcagaaaacaacctgaaccttccaaagatggctgaaaaagatgg 421 atgcttccaatctggattcaatgaggagacttgcctggtgaaaatcatcactggtctttt 481 ggagtttgaggtatacctagagtacctccagaacagatttgagagtagtgaggaacaagc 541 cagagctgtgcagatgagtacaaaagtcctgatccagttcctgcagaaaaaggcaaagaa 601 tctagatgcaataaccacccctgacccaaccacaaatgccagcctgctgacgaagctgca 661 ggcacagaaccagtggctgcaggacatgacaactcatctcattctgcgcagctttaagga 721 gttcctgcagtccagcctgagggctcttcggcaaatgtagcatgggcacctcagattgtt 781 gttgttaatgggcattccttcttctggtcagaaacctgtccactgggcacagaacttatg 841 ttgttctctatggagaactaaaagtatgagcgttaggacactattttaattatttttaat 901 ttattaatatttaaatatgtgaagctgagttaatttatgtaagtcatatttatattttta 961 agaagtaccacttgaaacattttatgtattagttttgaaataataatggaaagtggctat 1021 gcagtttgaatatcctttgtttcagagccagatcatttcttggaaagtgtaggcttacct 1081 caaataaatggctaacttatacatatttttaaagaaatatttatattgtatttatataat 1141 gtataaatggtttttataccaataaatggcattttaaaaaattcagcaaaaaaaaaa HumanIL-6mRNATranscript2 (SEQIDNO:55) 1 gtctcaatattagagtctcaacccccaataaatataggactggagatgtctgaggctcat 61 tctgccctcgagcccaccgggaacgaaagagaagctctatctcccctccaggagcccagc 121 tatgaactccttctccacaaacatgtaacaagagtaacatgtgtgaaagcagcaaagagg 181 cactggcagaaaacaacctgaaccttccaaagatggctgaaaaagatggatgcttccaat 241 ctggattcaatgaggagacttgcctggtgaaaatcatcactggtcttttggagtttgagg 301 tatacctagagtacctccagaacagatttgagagtagtgaggaacaagccagagctgtgc 361 agatgagtacaaaagtcctgatccagttcctgcagaaaaaggcaaagaatctagatgcaa 421 taaccacccctgacccaaccacaaatgccagcctgctgacgaagctgcaggcacagaacc 481 agtggctgcaggacatgacaactcatctcattctgcgcagctttaaggagttcctgcagt 541 ccagcctgagggctcttcggcaaatgtagcatgggcacctcagattgttgttgttaatgg 601 gcattccttcttctggtcagaaacctgtccactgggcacagaacttatgttgttctctat 661 ggagaactaaaagtatgagcgttaggacactattttaattatttttaatttattaatatt 721 taaatatgtgaagctgagttaatttatgtaagtcatatttatatttttaagaagtaccac 781 ttgaaacattttatgtattagttttgaaataataatggaaagtggctatgcagtttgaat 841 atcctttgtttcagagccagatcatttcttggaaagtgtaggcttacctcaaataaatgg 901 ctaacttatacatatttttaaagaaatatttatattgtatttatataatgtataaatggt 961 ttttataccaataaatggcattttaaaaaattcagcaaaaaaaaaa

Inhibitory Nucleic Acids

[0396] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL6R, sIL6gp130, or IL-6 protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0397] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein can be designed using the software available at the Integrated DNA Technologies website.

[0398] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0399] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an anti sense orientation to a target nucleic acid of interest).

[0400] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL6R, sIL6gp130, or IL-6 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0401] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, -units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0402] Exemplary antisense nucleic acids that are IL-6 receptor inhibitors are described in Keller et al., J. Immunol. 154(8):4091-4098, 1995; and Jiang et al., Anticancer Res. 31(9): 2899-2906, 2011.

[0403] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein (e.g., specificity for an IL6R, sIL6gp130, or IL-6 mRNA, e.g., specificity for any one of SEQ ID NOs: 50-55). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for an IL6R, sIL6gp130, or IL-6 mRNA can be designed based upon the nucleotide sequence of any of the IL6R, sIL6gp130, or IL-6 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL6R, sIL6gp130, or IL-6 mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, a SMAD7 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0404] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of an IL6R, sIL6gp130, or IL-6 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL6R, sIL6gp130, or IL-6 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0405] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Natl. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0406] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0407] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0408] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Natl. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0409] Another means by which expression of an IL6R, sIL6gp130, or IL-6 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL6R, sIL6gp130, or IL-6 polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0410] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0411] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0412] The siRNA molecules used to decrease expression of an IL6R, sIL6gp130, or IL-6 mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0413] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing an IL6R, sIL6gp130, or IL-6 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 50-55, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0414] Non-limiting examples of short interfering RNA (siRNA) that are IL-6 receptor inhibitors are described in Yi et al., Int. J. Oncol. 41(1):310-316, 2012; and Shinriki et al., Clin. Can. Res. 15(17):5426-5434, 2009). Non-limiting examples of microRNAs that are IL-6 receptor inhibitors are described in miR34a (Li et al., Int. J. Clin. Exp. Pathol. 8(2):1364-1373, 2015) and miR-451 (Liu et al., Cancer Epidemiol. 38(1):85-92, 2014).

[0415] Non-limiting examples of aptamers that are IL-6 receptor inhibitors are described in Meyer et al., RNA Biol. 11(1):57-65, 2014; Meyer et al., RNA Biol. 9(1):67-80, 2012; and Mittelberger et al., RNA Biol. 12(9):1043-1053, 2015. Additional examples of inhibitory nucleic acids that are IL-6 receptor inhibitors are described in, e.g., WO 96/040157.

[0416] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein can be administered to a subject (e.g., a human subject) in need thereof.

[0417] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0418] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0419] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL6R, sIL6gp130, or IL-6) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0420] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL6R, sIL6gp130, or IL-6) with a Tm of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive),

[0421] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0422] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0423] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0424] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0425] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0426] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0427] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0428] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0429] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0430] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0431] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0432] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0433] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0434] In some embodiments, the IL-6 receptor inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to IL-6. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to IL-6 receptor (e.g., one or both of IL6R and sIL6gp130).

[0435] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a V.sub.HH domain, a V.sub.NAR domain, a (scFv).sub.2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv.sub.2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab).sub.2-scFV.sub.2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0436] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab)2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0437] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0438] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of tocilizumab (artlizumab, Actemra; Sebba, Am. J. Health Syst. Pharm. 65(15):1413-1418, 2008; Tanaka et al., FEBS Letters 585(23):3699-3709, 2011; Nishimoto et al., Arthritis Rheum. 50:1761-1769, 2004; Yokota et al., Lancet 371(9617):998-1006, 2008; Emery et al., Ann. Rheum. Dis. 67(11):1516-1523, 2008; Roll et al., Arthritis Rheum. 63(5):1255-1264, 2011); lazakizumab (BMS945429; ALD518, a humanized monoclonal antibody that binds circulating IL-6 cytokine rather than the IL-6 receptor, blocking both classic signaling and trans-signaling (Weinblatt, Michael E., et al. The Efficacy and Safety of Subcutaneous Clazakizumab in Patients With Moderate-to-Severe Rheumatoid Arthritis and an Inadequate Response to Methotrexate: Results From a Multinational, Phase IIb, Randomized, Double-Blind, Placebo/Active-Controlled, Dose-Ranging Study. Arthritis & Rheumatology 67.10 (2015): 2591-2600.)); sarilumab (REGN88 or SAR153191; Huizinga et al., Ann. Rheum. Dis. 73(9):1626-1634, 2014; Sieper et al., Ann. Rheum. Dis. 74(6):1051-1057, 2014; Cooper, Immunotherapy 8(3): 249-250, 2016); MR-16 (Hartman et al., PLosOne 11(12):e0167195, 2016; Fujita et al., Biochim. Biophys. Acta. 10:3170-80, 2014; Okazaki et al., Immunol. Lett. 84(3):231-40, 2002; Noguchi-Sasaki et al., BMC Cancer 16:270, 2016; Ueda et al., Sci. Rep. 3:1196, 2013); rhPM-1 (MRA; Nishimoto et al., Blood 95: 56-61, 2000; Nishimoto et al., Blood 106: 2627-2632, 2005; Nakahara et al., Arthritis Rheum. 48(6): 1521-1529, 2003); NI-1201 (Lacroix et al., J. Biol. Chem. 290(45):26943-26953, 2015); EBI-029 (Schmidt et al., Eleven Biotherapeutics Poster # B0200, 2014). In some embodiments, the antibody is a nanobody (e.g., ALX-0061 (Van Roy et al., Arthritis Res. Ther. 17: 135, 2015; Kim et al., Arch. Pharm. Res. 38(5):575-584, 2015)). In some embodiments, the antibody is NRI or a variant thereof (Adachi et al., Mol. Ther. 11(1):5262-263, 2005; Hoshino et al., Can. Res. 67(3): 871-875, 2007). In some embodiments, the antibody is PF-04236921 (Pfizer) (Wallace et al., Ann. Rheum. Dis. 76(3):534-542, 2017).

[0439] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0440] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9 M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.5 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 1108 M, about 0.5108 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0441] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0442] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.Is.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.is.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.is.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins

[0443] In some embodiments, the IL-6 receptor inhibitor is a fusion protein, a soluble receptor, or a peptide (see e.g., U.S. Pat. No. 5,591,827). In some embodiments, the IL-6 receptor fusion protein comprises or consists of soluble gp130 (Jostock et al., Eur. J. Biochem. 268(1):160-167, 2001; Richards et al., Arthritis Rheum. 54(5):1662-1672, 2006; Rose-John et al., Exp. Opin. Ther. Targets 11(5):613-624, 2007).

[0444] In some embodiments, the IL-6 receptor fusion protein comprises or consists of FE999301 (Jostock et al., Eur. J. Biochem. 268(1):160-167, 2001) or sgp130Fc protein (Jones et al., J. Clin. Invest. 121(9):3375-3383, 2011). In some embodiments, the IL-6 receptor inhibitor is a peptide (e.g., S7 (Su et al., Cancer Res. 65(11):4827-4835, 2005). In some embodiments, the IL-6 receptor inhibitor is a triterpenoid saponin (e.g., chikusetsuaponin IVa butyl ester (CS-Iva-Be) (Yang et al., Mol. Cancer. Ther. 15(6):1190-200, 2016).

Small Molecules

[0445] In some embodiments, the IL-6 receptor inhibitor is a small molecule (see, e.g., U.S. Pat. No. 9,409,990). In some embodiments, the small molecule is LMT-28 (Hong et al., J. Immunol. 195(1): 237-245, 2015); ERBA (Enomoto et al., Biochem. Biophys. Res. Commun. 323:1096-1102, 2004; Boos et al., J. Nat. Prod. 75(4):661-668, 2012), ERBF (TB-2-081) (Hayashi et al., J. Pharmacol. Exp. Ther. 303:104-109, 2002; Vardanyan et al., Pain 151(2):257-265, 2010; Kino et al., J. Allergy Clin. Immunol. 120(2):437-444, 2007), or a variant thereof.

Immune Modulatory Agents

[0446] As used herein, the term immune modulatory agentomodifier refers to an agent that is a CD40/CD40 inhibitor (as defined herein), a CD3 inhibitor (as defined herein), a CD14 inhibitor (as defined agent), a CD20 inhibitor (as defined herein), a CD25 inhibitor (as defined herein), a CD28 inhibitor (as defined herein), a CD49 inhibitor (as defined herein), or a CD89 inhibitor. Examples of immune modulatory agents are described herein. Additional examples of immune modulatory agents are known in the art.

CD40/CD40L Inhibitors

[0447] The term CD40/CD40L inhibitors refers to an agent which decreases CD40 or CD40L (CD154) expression and/or the ability of CD40 to bind to CD40L (CD154). CD40 is a costimulatory receptor that binds to its ligand, CD40L (CD154).

[0448] In some embodiments, the CD40/CD40L inhibitor can decrease the binding between CD40 and CD40L by blocking the ability of CD40 to interact with CD40L. In some embodiments, the CD40/CD40L inhibitor can decrease the binding between CD40 and CD40L by blocking the ability of CD40L to interact with CD40. In some embodiments, the CD40/CD40L inhibitor decreases the expression of CD40 or CD40L. In some embodiments, the CD40/CD40L inhibitor decreases the expression of CD40. In some embodiments, the CD40/CD40L inhibitor decreases the expression of CD40L.

[0449] In some embodiments, the CD40/CD40L inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, or a small molecule. In some embodiments, the inhibitory nucleic acid is a small interfering RNA, an antisense nucleic acid, an aptamer, or a microRNA. Exemplary CD40/CD40L inhibitors are described herein. Additional examples of CD40/CD40L inhibitors are known in the art.

[0450] Exemplary aspects of different inhibitory nucleic acids are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of CD40 or CD40L mRNA in a mammalian cell can be synthesized in vitro. Inhibitory nucleic acids that can decrease the expression of CD40 or CD40L mRNA in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is 15 complementary to all or part of a CD40 or CD40L mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 56-61).

TABLE-US-00004 HumanCD40mRNA(Variant1)NM_001250.5 (SEQIDNO:56) 1 tttcctgggcggggccaaggctggggcaggggagtcagcagaggcctcgctcgggcgccc 61 agtggtcctgccgcctggtctcacctcgctatggttcgtctgcctctgcagtgcgtcctc 121 tggggctgcttgctgaccgctgtccatccagaaccacccactgcatgcagagaaaaacag 181 tacctaataaacagtcagtgctgttctttgtgccagccaggacagaaactggtgagtgac 241 tgcacagagttcactgaaacggaatgccttccttgcggtgaaagcgaattcctagacacc 301 tggaacagagagacacactgccaccagcacaaatactgcgaccccaacctagggcttcgg 361 gtccagcagaagggcacctcagaaacagacaccatctgcacctgtgaagaaggctggcac 421 tgtacgagtgaggcctgtgagagctgtgtcctgcaccgctcatgctcgcccggctttggg 481 gtcaagcagattgctacaggggtttctgataccatctgcgagccctgcccagtcggcttc 541 ttctccaatgtgtcatctgctttcgaaaaatgtcacccttggacaagctgtgagaccaaa 601 gacctggttgtgcaacaggcaggcacaaacaagactgatgttgtctgtggtccccaggat 661 cggctgagagccctggtggtgatccccatcatcttcgggatcctgtttgccatcctcttg 721 gtgctggtctttatcaaaaaggtggccaagaagccaaccaataaggccccccaccccaag 781 caggaaccccaggagatcaattttcccgacgatcttcctggctccaacactgctgctcca 841 gtgcaggagactttacatggatgccaaccggtcacccaggaggatggcaaagagagtcgc 901 atctcagtgcaggagagacagtgaggctgcacccacccaggagtgtggccacgtgggcaa 961 acaggcagttggccagagagcctggtgctgctgctgctgtggcgtgagggtgaggggctg 1021 gcactgactgggcatagctccccgcttctgcctgcacccctgcagtttgagacaggagac 1081 ctggcactggatgcagaaacagttcaccttgaagaacctctcacttcaccctggagccca 1141 tccagtctcccaacttgtattaaagacagaggcagaagtttggtggtggtggtgttgggg 1201 tatggtttagtaatatccaccagaccttccgatccagcagtttggtgcccagagaggcat 1261 catggtggcttccctgcgcccaggaagccatatacacagatgcccattgcagcattgttt 1321 gtgatagtgaacaactggaagctgcttaactgtccatcagcaggagactggctaaataaa 1381 attagaatatatttatacaacagaatctcaaaaacactgttgagtaaggaaaaaaaggca 1441 tgctgctgaatgatgggtatggaactttttaaaaaagtacatgcttttatgtatgtatat 1501 tgcctatggatatatgtataaatacaatatgcatcatatattgatataacaagggttctg 1561 gaagggtacacagaaaacccacagctcgaagagtggtgacgtctggggtggggaagaagg 1621 gtctggggg HumanCD40mRNA(Variant2)NM_152854.3 (SEQIDNO:57) 1 tttcctgggcggggccaaggctggggcaggggagtcagcagaggcctcgctcgggcgccc 61 agtggtcctgccgcctggtctcacctcgctatggttcgtctgcctctgcagtgcgtcctc 121 tggggctgcttgctgaccgctgtccatccagaaccacccactgcatgcagagaaaaacag 181 tacctaataaacagtcagtgctgttctttgtgccagccaggacagaaactggtgagtgac 241 tgcacagagttcactgaaacggaatgccttccttgcggtgaaagcgaattcctagacacc 301 tggaacagagagacacactgccaccagcacaaatactgcgaccccaacctagggcttcgg 361 gtccagcagaagggcacctcagaaacagacaccatctgcacctgtgaagaaggctggcac 421 tgtacgagtgaggcctgtgagagctgtgtcctgcaccgctcatgctcgcccggctttggg 481 gtcaagcagattgctacaggggtttctgataccatctgcgagccctgcccagtcggcttc 541 ttctccaatgtgtcatctgctttcgaaaaatgtcacccttggacaaggtccccaggatcg 601 gctgagagccctggtggtgatccccatcatcttcgggatcctgtttgccatcctcttggt 661 gctggtctttatcaaaaaggtggccaagaagccaaccaataaggccccccaccccaagca 721 ggaaccccaggagatcaattttcccgacgatcttcctggctccaacactgctgctccagt 781 gcaggagactttacatggatgccaaccggtcacccaggaggatggcaaagagagtcgcat 841 ctcagtgcaggagagacagtgaggctgcacccacccaggagtgtggccacgtgggcaaac 901 aggcagttggccagagagcctggtgctgctgctgctgtggcgtgagggtgaggggctggc 961 actgactgggcatagctccccgcttctgcctgcacccctgcagtttgagacaggagacct 1021 ggcactggatgcagaaacagttcaccttgaagaacctctcacttcaccctggagcccatc 1081 cagtctcccaacttgtattaaagacagaggcagaagtttggtggtggtggtgttggggta 1141 tggtttagtaatatccaccagaccttccgatccagcagtttggtgcccagagaggcatca 1201 tggtggcttccctgcgcccaggaagccatatacacagatgcccattgcagcattgtttgt 1261 gatagtgaacaactggaagctgcttaactgtccatcagcaggagactggctaaataaaat 1321 tagaatatatttatacaacagaatctcaaaaacactgttgagtaaggaaaaaaaggcatg 1381 ctgctgaatgatgggtatggaactttttaaaaaagtacatgcttttatgtatgtatattg 1441 cctatggatatatgtataaatacaatatgcatcatatattgatataacaagggttctgga 1501 agggtacacagaaaacccacagctcgaagagtggtgacgtctggggtggggaagaagggt 1561 ctggggg HumanCD40mRNA(Variant3)NM_001302753.1 (SEQIDNO:58) 1 tttcctgggcggggccaaggctggggcaggggagtcagcagaggcctcgctcgggcgccc 61 agtggtcctgccgcctggtctcacctcgctatggttcgtctgcctctgcagtgcgtcctc 121 tggggctgcttgctgaccgctgtccatccagaaccacccactgcatgcagagaaaaacag 181 tacctaataaacagtcagtgctgttctttgtgccagccaggacagaaactggtgagtgac 241 tgcacagagttcactgaaacggaatgccttccttgcggtgaaagcgaattcctagacacc 301 tggaacagagagacacactgccaccagcacaaatactgcgaccccaacctagggcttcgg 361 gtccagcagaagggcacctcagaaacagacaccatctgcacctgtgaagaaggctggcac 421 tgtacgagtgaggcctgtgagagctgtgtcctgcaccgctcatgctcgcccggctttggg 481 gtcaagcagattgctacaggggtttctgataccatctgcgagccctgcccagtcggcttc 541 ttctccaatgtgtcatctgctttcgaaaaatgtcacccttggacaagctgtgagaccaaa 601 gacctggttgtgcaacaggcaggcacaaacaagactgatgttgtctgtggtgagtcctgg 661 acaatgggccctggagaaagcctaggaaggtccccaggatcggctgagagccctggtggt 721 gatccccatcatcttcgggatcctgtttgccatcctcttggtgctggtctttatcaaaaa 781 ggtggccaagaagccaaccaataaggccccccaccccaagcaggaaccccaggagatcaa 841 ttttcccgacgatcttcctggctccaacactgctgctccagtgcaggagactttacatgg 901 atgccaaccggtcacccaggaggatggcaaagagagtcgcatctcagtgcaggagagaca 961 gtgaggctgcacccacccaggagtgtggccacgtgggcaaacaggcagttggccagagag 1021 cctggtgctgctgctgctgtggcgtgagggtgaggggctggcactgactgggcatagctc 1081 cccgcttctgcctgcacccctgcagtttgagacaggagacctggcactggatgcagaaac 1141 agttcaccttgaagaacctctcacttcaccctggagcccatccagtctcccaacttgtat 1201 taaagacagaggcagaagtttggtggtggtggtgttggggtatggtttagtaatatccac 1261 cagaccttccgatccagcagtttggtgcccagagaggcatcatggtggcttccctgcgcc 1321 caggaagccatatacacagatgcccattgcagcattgtttgtgatagtgaacaactggaa 1381 gctgcttaactgtccatcagcaggagactggctaaataaaattagaatatatttatacaa 1441 cagaatctcaaaaacactgttgagtaaggaaaaaaaggcatgctgctgaatgatgggtat 1501 ggaactttttaaaaaagtacatgcttttatgtatgtatattgcctatggatatatgtata 1561 aatacaatatgcatcatatattgatataacaagggttctggaagggtacacagaaaaccc 1621 acagctcgaagagtggtgacgtctggggtggggaagaagggtctggggg HumanCD40mRNA(Variant5)NM_001322421.1 (SEQIDNO:59) 1 tttcctgggcggggccaaggctggggcaggggagtcagcagaggcctcgctcgggcgccc 61 agtggtcctgccgcctggtctcacctcgctatggttcgtctgcctctgcagtgcgtcctc 121 tggggctgcttgctgaccgctgtccatccagaaccacccactgcatgcagagaaaaacag 181 tacctaataaacagtcagtgctgttctttgtgccagccaggacagaaactggtgagtgac 241 tgcacagagttcactgaaacggaatgccttccttgcggtgaaagcgaattcctagacacc 301 tggaacagagagacacactgccaccagcacaaatactgcgaccccaacctagggcttcgg 361 gtccagcagaagggcacctcagaaacagacaccatctgcacctgtgaagaaggctggcac 421 tgtacgagtgaggcctgtgagagctgtgtcctgcaccgctcatgctcgcccggctttggg 481 gtcaagcagattgctacaggggtttctgataccatctgcgagccctgcccagtcggcttc 541 ttctccaatgtgtcatctgctttcgaaaaatgtcacccttggacaagctgtgagaccaaa 601 gacctggttgtgcaacaggcaggcacaaacaagactgatgttgtctgtggtccccaggat 661 cggctgagagccctggtggtgatccccatcatcttcgggatcctgtttgccatcctcttg 721 gtgctggtctttatcagtgagtcctcagaaaaggtggccaagaagccaaccaataaggcc 781 ccccaccccaagcaggaaccccaggagatcaattttcccgacgatcttcctggctccaac 841 actgctgctccagtgcaggagactttacatggatgccaaccggtcacccaggaggatggc 901 aaagagagtcgcatctcagtgcaggagagacagtgaggctgcacccacccaggagtgtgg 961 ccacgtgggcaaacaggcagttggccagagagcctggtgctgctgctgctgtggcgtgag 1021 ggtgaggggctggcactgactgggcatagctccccgcttctgcctgcacccctgcagttt 1081 gagacaggagacctggcactggatgcagaaacagttcaccttgaagaacctctcacttca 1141 ccctggagcccatccagtctcccaacttgtattaaagacagaggcagaagtttggtggtg 1201 gtggtgttggggtatggtttagtaatatccaccagaccttccgatccagcagtttggtgc 1261 ccagagaggcatcatggtggcttccctgcgcccaggaagccatatacacagatgcccatt 1321 gcagcattgtttgtgatagtgaacaactggaagctgcttaactgtccatcagcaggagac 1381 tggctaaataaaattagaatatatttatacaacagaatctcaaaaacactgttgagtaag 1441 gaaaaaaaggcatgctgctgaatgatgggtatggaactttttaaaaaagtacatgctttt 1501 atgtatgtatattgcctatggatatatgtataaatacaatatgcatcatatattgatata 1561 acaagggttctggaagggtacacagaaaacccacagctcgaagagtggtgacgtctgggg 1621 tggggaagaagggtctggggg HumanCD40mRNA(Variant6)NM_001322422.1 (SEQIDNO:60) 1 tttcctgggcggggccaaggctggggcaggggagtcagcagaggcctcgctcgggcgccc 61 agtggtcctgccgcctggtctcacctcgctatggttcgtctgcctctgcagtgcgtcctc 121 tggggctgcttgctgaccgctgtccatccagaaccacccactgcatgcagagaaaaacag 181 tacctaataaacagtcagtgctgttctttgtgccagccaggacagaaactggtgagtgac 241 tgcacagagttcactgaaacggaatgccttccttgcggtgaaagcgaattcctagacacc 301 tggaacagagagacacactgccaccagcacaaatactgcgaccccaacctagggcttcgg 361 gtccagcagaagggcacctcagaaacagacaccatctgcacctgtgaagaaggctggcac 421 tgtacgagtgaggcctgtgagagctgtgtcctgcaccgctcatgctcgcccggctttggg 481 gtcaagcagattggtccccaggatcggctgagagccctggtggtgatccccatcatcttc 541 gggatcctgtttgccatcctcttggtgctggtctttatcaaaaaggtggccaagaagcca 601 accaataaggccccccaccccaagcaggaaccccaggagatcaattttcccgacgatctt 661 cctggctccaacactgctgctccagtgcaggagactttacatggatgccaaccggtcacc 721 caggaggatggcaaagagagtcgcatctcagtgcaggagagacagtgaggctgcacccac 781 ccaggagtgtggccacgtgggcaaacaggcagttggccagagagcctggtgctgctgctg 841 ctgtggcgtgagggtgaggggctggcactgactgggcatagctccccgcttctgcctgca 901 cccctgcagtttgagacaggagacctggcactggatgcagaaacagttcaccttgaagaa 961 cctctcacttcaccctggagcccatccagtctcccaacttgtattaaagacagaggcaga 1021 agtttggtggtggtggtgttggggtatggtttagtaatatccaccagaccttccgatcca 1081 gcagtttggtgcccagagaggcatcatggtggcttccctgcgcccaggaagccatataca 1141 cagatgcccattgcagcattgtttgtgatagtgaacaactggaagctgcttaactgtcca 1201 tcagcaggagactggctaaataaaattagaatatatttatacaacagaatctcaaaaaca 1261 ctgttgagtaaggaaaaaaaggcatgctgctgaatgatgggtatggaactttttaaaaaa 1321 gtacatgcttttatgtatgtatattgcctatggatatatgtataaatacaatatgcatca 1381 tatattgatataacaagggttctggaagggtacacagaaaacccacagctcgaagagtgg 1441 tgacgtctggggtggggaagaagggtctggggg HumanCD154(CD40L)mRNANM_000074.2 (SEQIDNO:61) 1 actttgacagtcttctcatgctgcctctgccaccttctctgccagaagataccatttcaa 61 ctttaacacagcatgatcgaaacatacaaccaaacttctccccgatctgcggccactgga 121 ctgcccatcagcatgaaaatttttatgtatttacttactgtttttcttatcacccagatg 181 attgggtcagcactttttgctgtgtatcttcatagaaggttggacaagatagaagatgaa 241 aggaatcttcatgaagattttgtattcatgaaaacgatacagagatgcaacacaggagaa 301 agatccttatccttactgaactgtgaggagattaaaagccagtttgaaggctttgtgaag 361 gatataatgttaaacaaagaggagacgaagaaagaaaacagctttgaaatgcaaaaaggt 421 gatcagaatcctcaaattgcggcacatgtcataagtgaggccagcagtaaaacaacatct 481 gtgttacagtgggctgaaaaaggatactacaccatgagcaacaacttggtaaccctggaa 541 aatgggaaacagctgaccgttaaaagacaaggactctattatatctatgcccaagtcacc 601 ttctgttccaatcgggaagcttcgagtcaagctccatttatagccagcctctgcctaaag 661 tcccccggtagattcgagagaatcttactcagagctgcaaatacccacagttccgccaaa 721 ccttgcgggcaacaatccattcacttgggaggagtatttgaattgcaaccaggtgcttcg 781 gtgtttgtcaatgtgactgatccaagccaagtgagccatggcactggcttcacgtccttt 841 ggcttactcaaactctgaacagtgtcaccttgcaggctgtggtggagctgacgctgggag 901 tcttcataatacagcacagcggttaagcccaccccctgttaactgcctatttataaccct 961 aggatcctccttatggagaactatttattatacactccaaggcatgtagaactgtaataa 1021 gtgaattacaggtcacatgaaaccaaaacgggccctgctccataagagcttatatatctg 1081 aagcagcaaccccactgatgcagacatccagagagtcctatgaaaagacaaggccattat 1141 gcacaggttgaattctgagtaaacagcagataacttgccaagttcagttttgtttctttg 1201 cgtgcagtgtctttccatggataatgcatttgatttatcagtgaagatgcagaagggaaa 1261 tggggagcctcagctcacattcagttatggttgactctgggttcctatggccttgttgga 1321 gggggccaggctctagaacgtctaacacagtggagaaccgaaaccccccccccccccccg 1381 ccaccctctcggacagttattcattctctttcaatctctctctctccatctctctctttc 1441 agtctctctctctcaacctctttcttccaatctctctttctcaatctctctgtttccctt 1501 tgtcagtctcttccctcccccagtctctcttctcaatccccctttctaacacacacacac 1561 acacacacacacacacacacacacacacacacacacacacagagtcaggccgttgctagt 1621 cagttctcttctttccaccctgtccctatctctaccactatagatgagggtgaggagtag 1681 ggagtgcagccctgagcctgcccactcctcattacgaaatgactgtatttaaaggaaatc 1741 tattgtatctacctgcagtctccattgtttccagagtgaacttgtaattatcttgttatt 1801 tattttttgaataataaagacctcttaacattaa

Inhibitory Nucleic Acids

[0451] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a CD40 or CD40L protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0452] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a CD40 or CD40L protein described herein. Antisense nucleic acids targeting a nucleic acid encoding a CD40 or CD40L protein can be designed using the software available at the Integrated DNA Technologies website.

[0453] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine-substituted nucleotides can be used.

[0454] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0455] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human, using any of the devices described herein. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a CD40 or CD40L protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0456] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 0-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0457] Some exemplary antisense nucleic acids that are CD40 or CD40L inhibitors are described, e.g., in U.S. Pat. Nos. 6,197,584 and 7,745,609; Gao et al., Gut 54(1):70-77, 2005; Arranz et al., J Control Release 165(3):163-172, 2012; Donner et al., Mol. Ther. Nucleic Acids 4:e265, 2015.

[0458] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a CD40 or CD40L protein (e.g., specificity for a CD40 or CD40L mRNA, e.g., specificity for any one of SEQ ID NOs: 56-61). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a CD40 or CD40L mRNA can be designed based upon the nucleotide sequence of any of the CD40 or CD40L mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a CD40 or CD40L mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, a CD40 or CD40L mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0459] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a CD40 or CD40L polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the CD40 or CD40L polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0460] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorg. Med. Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Nat. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0461] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0462] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorg. Med. Chem. Lett. 5:1119-11124, 1975).

[0463] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Nat. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Nat. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0464] Another means by which expression of a CD40 or CD40L mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a CD40 or CD40L polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0465] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Nat. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0466] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0467] The siRNA molecules used to decrease expression of a CD40 or CD40L mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0468] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing a CD40 or CD40L mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 56-61, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0469] Non-limiting examples of short interfering RNA (siRNA) that are CD40/CD40L inhibitors are described in, e.g., Pluvinet et al., Blood 104:3642-3646, 2004; Karimi et al., Cell Immunol. 259(1):74-81, 2009; and Zheng et al., Arthritis Res. Ther. 12(1):R13, 2010. Non-limiting examples of short hairpin RNA (shRNA) targeting CD40/CD40L are described in Zhang et al., Gene Therapy 21:709-714, 2014. Non-limiting examples of microRNAs that are CD40/CD40L inhibitors include, for example, miR146a (Chen et al., FEBS Letters 585(3):567-573, 2011), miR-424, and miR-503 (Lee et al., Sci. Rep. 7:2528, 2017).

[0470] Non-limiting examples of aptamers that are CD40/CD40L inhibitors are described in Soldevilla et al., Biomaterials 67:274-285, 2015.

[0471] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a CD40 or CD40L protein can be delivered locally to a subject (e.g., a human subject) in need thereof using any of the devices described herein.

[0472] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0473] Any of the inhibitor nucleic acids described herein can be formulated for administration to the gastrointestinal tract. See, e.g., the formulation methods described in US 2016/0090598 and Schoellhammer et al., Gastroenterology, doi: 10.1053/j.gastro.2017.01.002, 2017.

[0474] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stringent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0475] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding CD40 or CD40L) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0476] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding CD40 or CD40L) with a Tm of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive).

[0477] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0478] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0479] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0480] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0481] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0482] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0483] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0484] In certain embodiments, pharmaceutical compositions provided herein can include one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, and polyvinylpyrrolidone.

[0485] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents, such as dimethylsulfoxide, are used.

[0486] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0487] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose. Any of the pharmaceutical compositions described herein can be delivered locally to a subject using any of the devices described herein.

[0488] In some examples, an inhibitory nucleic acid can be formulated to include a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, an inhibitory nucleic acid can be formulated as a suspension and can be prepared using appropriate liquid carriers, suspending agents, and the like. An inhibitory nucleic acid can be formulated as a suspension, solution, or emulsion in oily or aqueous vehicles prior to intrathecal administration using any of the devices described herein, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for formulating an inhibitory nucleic acid include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0489] In some embodiments, the CD40/CD40L inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CD40 or CD40L, or to both CD40 and CD40L.

[0490] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0491] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0492] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0493] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0494] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of PG102 (Pangenetics) (Bankert et al., J. Immunol. 194(9):4319-4327, 2015); 2C10 (Lowe et al., Am. J. Transplant 12(8):2079-2087, 2012); ASKP1240 (Bleselumab) (Watanabe et al., Am. J. Transplant 13(8):1976-1988, 2013); 4D11 (Imai et al., Transplantation 84(8):1020-1028, 2007); BI 655064 (Boehringer Ingelheim) (Visvanathan et al., 2016 American College of Rheumatology Annual Meeting, Abstract 1588, Sep. 28, 2016); 5D12 (Kasran et al., Aliment. Pharmacol. Ther., 22(2):111-122, 2005; Boon et al., Toxicology 174(1):53-65, 2002); ruplizumab (hu5c8) (Kirk et al., Nat. Med. 5(6):686-693, 1999); CHIR12.12 (HCD122) (Weng et al., Blood 104(11):3279, 2004; Tai et al., Cancer Res. 65(13):5898-5906, 2005); CDP7657 (Shock et al., Arthritis Res. Ther. 17(1):234, 2015); BMS-986004 domain antibody (dAb) (Kim et al., Am. J. Transplant. 17(5):1182-1192, 2017); 5c8 (Xie et al., J Immunol. 192(9):4083-4092, 2014); dacetuzumab (SGN-40) (Lewis et al., Leukemia 25(6):1007-1016, 2011; and Khubchandani et al., Curr. Opin. Investig. Drugs 10(6):579-587, 2009); lucatumumab (HCD122) (Bensinger et al., Br. J. Haematol. 159: 58-66, 2012; and Byrd et al., Leuk. Lymphoma 53(11): 10.3109/10428194.2012.681655, 2012); PG102 (FFP104) (Bankert et al., J. Immunol. 194(9):4319-4327, 2015); Chi Lob 7/4 (Johnson et al., J. Clin. Oncol. 28:2507, 2019); and ASKP1240 (Okimura et al., Am. J. Transplant. 14(6): 1290-1299, 2014; or Ma et al., Transplantation 97(4): 397-404, 2014).

[0495] Further teachings of CD40/CD40L antibodies and antigen-binding fragments thereof are described in, for example, U.S. Pat. Nos. 5,874,082; 7,169,389; 7,271,152; 7,288,252; 7,445,780; 7,537,763, 8,277,810; 8,293,237, 8,551,485; 8,591,900; 8,647,625; 8,784,823; 8,852,597; 8,961,976; 9,023,360, 9,028,826; 9,090,696, 9,221,913; US2014/0093497; and US2015/0017155 each of which is incorporated by reference in its entirety.

[0496] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0497] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0498] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0499] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.is.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.is.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.is.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion and Truncated Proteins and Peptides

[0500] In some embodiments, the CD40/CD40L inhibitor is a fusion protein, a truncated protein (e.g., a soluble receptor) or a peptide. In some embodiments, the CD40/CD40L inhibitor is a truncated protein as disclosed in, for example, WO 01/096397. In some embodiments, the CD40/CD40L inhibitor is a peptide, such as a cyclic peptide (see, e.g., U.S. Pat. No. 8,802,634; Bianco et al., Org. Biomol. Chem. 4:1461-1463, 2006; Deambrosis et al., J. Mol. Med. 87(2):181-197, 2009; Vaitaitis et al., Diabetologia 57(11):2366-2373, 2014). In some embodiments, the CD40/CD40L inhibitor is a CD40 ligand binder, for example, a Tumor Necrosis Factor Receptor-associated Factor (TRAF): TRAF2, TRAF3, TRAF6, TRAF5 and TTRAP, or E3 ubiquitin-protein ligase RNF128.

Small Molecules

[0501] In some embodiments, the CD40/CD40L inhibitor is a small molecule (see, e.g., U.S. Pat. No. 7,173,046, U.S. Patent Application No. 2011/0065675). In some embodiments, the small molecule is Bio8898 (Silvian et al., ACS Chem. Biol. 6(6):636-647, 2011); Suramin (Margolles-Clark et al., Biochem. Pharmacol. 77(7):1236-1245, 2009); a small-molecule organic dye (Margolles-Clark et al., J. Mol. Med. 87(11):1133-1143, 2009; Buchwald et al., J. Mol. Recognit. 23(1):65-73, 2010), a naphthalenesulphonic acid derivative (Margolles-Clark et al., Chem. Biol. Drug Des. 76(4):305-313, 2010), or a variant thereof.

CD3 Inhibitors

[0502] The term CD3 inhibitor refers to an agent which decreases the ability of one or more of CD3, CD3, CD3, and CD3 to associate with one or more of TCR-, TCR-, TCR-, and TCR-. In some embodiments, the CD3 inhibitor can decrease the association between one or more of CD3, CD3, CD3, and CD3 and one or more of TCR-, TCR-, TCR-, and TCR- by blocking the ability of one or more of CD3, CD3, CD3, and CD3 to interact with one or more of TCR-, TCR-, TCR-, and TCR-.

[0503] In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment thereof, a fusion protein, or a small molecule. Exemplary CD3 inhibitors are described herein. Additional examples of CD3 inhibitors are known in the art.

[0504] Exemplary sequences for human CD3, human CD3, human CD3, and human CD3 are shown below.

TABLE-US-00005 HumanCD3 (SEQIDNO:62) meqgkglavlilaiillqgtlaqsikgnhlvkvydyqedg sylltcdaeaknitwfkdgkmigfltedkkkwnlgsnakd prgmyqckgsqnkskplqvyyrmcqncielnaatisgflf aeivsifvlavgvyfiagqdgvrqsrasdkqtllpndqly qplkdreddqyshlqgnqlrrn HumanCD3IsoformA (SEQIDNO:63) fkipieeledrvfvncntsitwvegtvgtllsditrldl gkrildprgiyrcngtdiykdkestvqvhyrmcqscveld patvagiivtdviatlllalgvfcfaghetgrlsgaadtq allrndqvyqplrdrddaqyshlggnwarnk HumanCD3IsoformB (SEQIDNO:64) fkipieeledrvfvncntsitwvegtvgtllsditrldl gkrildprgiyrcngtdiykdkestvqvhyrtadtqallr ndqvyqpirdrddaqyshlggnwarnk HumanCD3 (SEQIDNO:65) dgneemggitqtpykvsisgttviltcpqypgseilwq hndkniggdeddknigsdedhlslkefseleqsgyyvcyp rgskpedanfylylrarvcencmemdvmsvativivdici tggllllvyywsknrkakakpvtrgagaggrqrgqnkerp ppvpnpdyepirkgqrdlysglnqrri HumanCD3Isoform1 (SEQIDNO:66) qsfglldpklcylldgilfiygviltalflrvkfsrsad apayqqgqnqlynelnlgrreeydvldkrrgrdpemggkp qrrknpqeglynelqkdkmaeayseigmkgerrrgkghdg lyqglstatkdtydalhmqalppr HumanCD3Isoform2 (SEQIDNO:67) qsfglldpklcylldgilfiygviltalflrvkfsrsad apayqqgqnqlynelnlgrreeydvldkrrgrdpemggkp rrknpqeglynelqkdkmaeayseigmkgerrrgkghdgl yqglstatkdtydalhmqalppr

Antibodies

[0505] In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3. In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3. In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3. In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3. In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment that can bind to two or more (e.g., two, three, or four) of CD3, CD3, CD3, and CD3.

[0506] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0507] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0508] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0509] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0510] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of visiluzumab (Nuvion; HuM-291; M291; SMART anti-CD3 antibody) (Carpenter et al., Biol. Blood Marrow Transplant 11(6): 465-471, 2005; Trajkovic Curr. Opin. Investig. Drugs 3(3): 411-414, 2002; Malviya et al., J. Nucl. Med. 50(10): 1683-1691, 2009); muromonab-CD3 (orthoclone OKT3) (Hori et al., Surg. Today 41(4): 585-590, 2011; Norman Ther. Drug Monit. 17(6): 615-620, 1995; and Gramatzki et al., Leukemia 9(3): 382-390, 19); otelixizumab (TRX4) (Vossenkamper et al., Gastroenterology 147(1): 172-183, 2014; and Wiczling et al., J. Clin. Pharmacol. 50(5): 494-506, 2010); foralumab (NI-0401) (Ogura et al., Clin. Immunol. 183: 240-246; and van der Woude et al., Inflamm. Bowel Dis. 16: 1708-1716, 2010); ChAgly CD3; teplizumab (MGA031) (Waldron-Lynch et al., Sci. Transl. Med. 4(118): 118ra12, 2012; and Skelley et al., Ann. Pharmacother 46(10): 1405-1412, 2012); or catumaxomab (Removab) (Linke et al., Mabs 2(2): 129-136, 2010; and Bokemeyer et al., Gastric Cancer 18(4): 833-842, 2015).

[0511] Additional examples of CD3 inhibitors that are antibodies or antibody fragments are described in, e.g., U.S. Patent Application Publication Nos. 2017/0204194, 2017/0137519, 2016/0368988, 2016/0333095, 2016/0194399, 2016/0168247, 2015/0166661, 2015/0118252, 2014/0193399, 2014/0099318, 2014/0088295, 2014/0080147, 2013/0115213, 2013/0078238, 2012/0269826, 2011/0217790, 2010/0209437, 2010/0183554, 2008/0025975, 2007/0190045, 2007/0190052, 2007/0154477, 2007/0134241, 2007/0065437, 2006/0275292, 2006/0269547, 2006/0233787, 2006/0177896, 2006/0165693, 2006/0088526, 2004/0253237, 2004/0202657, 2004/0052783, 2003/0216551, and 2002/0142000, each of which is herein incorporated by reference in its entirety (e.g., the sections describing the CD3 inhibitors). Additional CD3 inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., Smith et al., J. Exp. Med. 185(8):1413-1422, 1997; Chatenaud et al., Nature 7:622-632, 2007.

[0512] In some embodiments, the CD3 inhibitor comprises or consists of a bispecific antibody (e.g., JNJ-63709178) (Gaudet et al., Blood 128(22): 2824, 2016); JNJ-64007957 (Girgis et al., Blood 128: 5668, 2016); MGDO09 (Tolcher et al., J. Clin. Oncol. 34:15, 2016); ERY974 (Ishiguro et al., Sci. Transl. Med. 9(410): pii.eaa4291, 2017); AMV564 (Hoseini and Cheung Blood Cancer J. 7:e522, 2017); AFM11 (Reusch et al., MAbs 7(3): 584-604, 2015); duvortuxizumab (JNJ 64052781); R06958688; blinatumomab (Blincyto; AMG103) (Ribera Expert Rev. Hematol. 1:1-11, 2017; and Mori et al., N Engl. J. Med. 376(23):e49, 2017); XmAb13676; or REGN1979 (Bannerji et al., Blood 128: 621, 2016; and Smith et al., Sci. Rep. 5:17943, 2015)).

[0513] In some embodiments, the CD3 inhibitor comprises or consists of a trispecific antibody (e.g., ertumaxomab (Kiewe and Thiel, Expert Opin. Investig. Drugs 17(10): 1553-1558, 2008; and Haense et al., BMC Cancer 16:420, 2016); or FBTA05 (Bi20; Lymphomun) (Buhmann et al., J. Transl. Med. 11:160, 2013; and Schuster et al., Br. J Haematol. 169(1): 90-102, 2015)).

[0514] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0515] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0516] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0517] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion and Truncated Proteins and Peptides

[0518] In some embodiments, the CD3 inhibitor is a fusion protein, a truncated protein (e.g., a soluble receptor), or a peptide. In some embodiments, the CD3 inhibitor can be a fusion protein (see, e.g., Lee et al., Oncol. Rep. 15(5): 1211-1216, 2006).

Small Molecules

[0519] In some embodiments, the CD3 inhibitor comprises or consists of a bispecific small molecule-antibody conjugate (see, e.g., Kim et al., PNAS 110(44): 17796-17801, 2013; Viola et al., Eur. J. Immunol. 27(11):3080-3083, 1997).

CD14 Inhibitors

[0520] The term CD14 inhibitors refers to an agent which decreases the ability of CD14 to bind to lipopolysaccharide (LPS). CD14 acts as a co-receptor with Toll-like receptor 4 (TLR4) that binds LPS in the presence of lipopolysaccharide-binding protein (LBP). In some embodiments, the CD14 inhibitor can decrease the binding between CD14 and LPS by blocking the ability of CD14 to interact with LPS.

[0521] In some embodiments, the CD14 inhibitor is an antibody or an antigen-binding fragment thereof. In some embodiments, the CD14 inhibitor is a small molecule. Exemplary CD14 inhibitors are described herein. Additional examples of CD14 inhibitors are known in the art.

[0522] An exemplary sequence for human CD14 is shown below.

TABLE-US-00006 HumanCD14 (SEQIDNO:68) maaaaasrgvgaklglreirihlcqrspgsqgvrdfiekr yvelkkanpdlpilirecsdvqpklwaryafgqetnvpin nfsadqvtralenvlsgka

CD14 InhibitorsAntibodies

[0523] In some embodiments, the CD14 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, the CD14 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD14.

[0524] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0525] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0526] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0527] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0528] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of IC14 (Axtelle and Pribble, J. Endotoxin Res. 7(4): 310-314, 2001; Reinhart et al., Crit. Care Med. 32(5): 1100-1108, 2004; Spek et al., J. Clin. Immunol. 23(2): 132-140, 2003). Additional examples of anti-CD14 antibodies and CD14 inhibitors can be found, e.g., in WO 2015/140591 and WO 2014/122660, incorporated in its entirety herein.

[0529] Additional examples of CD14 inhibitors that are antibodies or antibody fragments are described in, e.g., U.S. Patent Application Serial No. 2017/0107294, 2014/0050727, 2012/0227412, 2009/0203052, 2009/0029396, 2008/0286290, 2007/0106067, 2006/0257411, 2006/0073145, 2006/0068445, 2004/0092712, 2004/0091478, and 2002/0150882, each of which is herein incorporated by reference (e.g., the sections that describe CD14 inhibitors).

[0530] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0531] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0532] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0533] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 Ms.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0534] Additional examples of CD14 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD14 InhibitorsSmall Molecules

[0535] In some embodiments, the CD14 inhibitor is a small molecule. Non-limiting examples of CD14 inhibitors that are small molecules are described in, e.g., methyl 6-deoxy-6-N-dimethyl-N-cyclopentylammonium-2, 3-di-O-tetradecyl--D-glucopyranoside iodide (IAXO-101); methyl 6-Deoxy-6-amino-2,3-di-O-tetradecyl--D-glucopyranoside (IAXO-102); N-(3,4-bis-tetradecyloxy-benzyl)-N-cyclopentyl-N,N-dimethylammonium iodide (IAXO-103); and IMO-9200.

[0536] Additional examples of CD14 inhibitors that are small molecules are known in the art.

CD20 Inhibitors

[0537] The term CD20 inhibitors refers to an agent that binds specifically to CD20 expressed on the surface of a mammalian cell.

[0538] In some embodiments, the CD20 inhibitor is an antibody or an antigen-binding fragment thereof, or a fusion protein or peptide. Exemplary CD20 inhibitors are described herein. Additional examples of CD20 inhibitors are known in the art.

[0539] An exemplary sequence of human CD20 is shown below.

TABLE-US-00007 HumanCD20 (SEQIDNO:69) mttprnsvngtfpaepmkgpiamqsgpkplfrrmsslvgp tqsffmresktlgavqimnglfhialggllmipagiyapi cvtvwyplwggimyiisgsllaateknsrkclvkgkmimn slslfaaisgmilsimdilnikishflkmeslnfirahtp yiniyncepanpseknspstqycysiqslflgilsvmlif affqelviagivenewkrtcsrpksnivilsaeekkeqti eikeevvgltetssqpkneedieiipiqeeeeeetetnfp eppqdqesspiendssp

CD20 InhibitorsAntibodies

[0540] In some embodiments, the CD20 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).

[0541] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0542] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0543] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0544] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0545] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of rituximab (Rituxan, MabThera, MK-8808) (Ji et al., Indian J. Hematol. Blood Transfus. 33(4): 525-533, 2017; and Calderon-Gomez and Panes Gastroenterology 142(1): 1741-76, 2012); -PF-05280586; ocrelizumab (Ocrevus) (Sharp N. Engl. J. Med. 376(17): 1692, 2017); of atumumab (Arzerra; HuMax-CD20) (ADallal Ther. Clin. Risk Manag. 13:905-907, 2017; and Furman et al., Lancet Haematol. 4(1): e24-e34, 2017); PF-05280586 (Williams et al., Br. J Clin. Pharmacol. 82(6): 1568-1579, 2016; and Cohen et al., Br. J. Clin. Pharmacol. 82(1): 129-138, 2016); obinutuzumab (Gazyva) (Reddy et al., Rheumatology 56(7): 1227-1237, 2017; and Marcus et al., N. Engl. J. Med. 377(14): 1331-1344, 2017); ocaratuzumab (AME-133v; LY2469298) (Cheney et al., Mabs 6(3): 749-755, 2014; and Tobinai et al., Cancer Sci. 102(2): 432-8, 2011); GP2013 (Jurczak et al., Lancet Haenatol. 4(8): e350-e361, 2017); IBI301; HLX01; veltuzumab (hA20) (Kalaycio et al., Leuk. Lymphoma 57(4): 803-811, 2016; and Ellebrecht et al., JAMA Dermatol. 150(12): 1331-1335, 2014); SCT400 (Gui et al., Chin. J. Cancer Res. 28(2): 197-208); ibritumomab tiuxetan (Zevalin) (Philippe et al., Bone Marrow Transplant 51(8): 1140-1142, 2016; and Lossos et al., Leuk. Lymphoma 56(6): 1750-1755, 2015); ublituximab (TG1101) (Sharman et al., Blood 124: 4679, 2014; and Sawas et al., Br. J. Haematol. 177(2): 243-253, 2017); LFB-R603 (Esteves et al., Blood 118: 1660, 2011; and Baritaki et al., Int. J. Oncol. 38(6): 1683-1694, 2011); or tositumomab (Bexxar) (Buchegger et al., J. Nucl. Med. 52(6): 896-900, 2011; and William and Bierman Expert Opin. Biol. Ther. 10(8): 1271-1278, 2010). Additional examples of CD20 antibodies are known in the art (see, e.g., WO 2008/156713).

[0546] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of a bispecific antibody (e.g., XmAb13676; REGN1979 (Bannerji et al., Blood 128: 621, 2016; and Smith et al., Sci. Rep. 5: 17943, 2015); PRO131921 (Casulo et al., Clin. Immnol. 154(1): 37-46, 2014; and Robak and Robak BioDrugs 25(1): 13-25, 2011); or Acellbia).

[0547] In some embodiments, the CD20 inhibitor comprises or consists of a trispecific antibody (e.g., FBTA05 (Bi20; Lymphomun) (Buhmann et al., J. Transl. Med. 11:160, 2013; and Schuster et al., Br. J Haematol. 169(1): 90-102, 2015)).

[0548] Additional examples of CD20 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos. 2017/0304441, 2017/0128587, 2017/0088625, 2017/0037139, 2017/0002084, 2016/0362472, 2016/0347852, 2016/0333106, 2016/0271249, 2016/0243226, 2016/0115238, 2016/0108126, 2016/0017050, 2016/0017047, 2016/0000912, 2016/0000911, 2015/0344585, 2015/0290317, 2015/0274834, 2015/0265703, 2015/0259428, 2015/0218280, 2015/0125446, 2015/0093376, 2015/0079073, 2015/0071911, 2015/0056186, 2015/0010540, 2014/0363424, 2014/0356352, 2014/0328843, 2014/0322200, 2014/0294807, 2014/0248262, 2014/0234298, 2014/0093454, 2014/0065134, 2014/0044705, 2014/0004104, 2014/0004037, 2013/0280243, 2013/0273041, 2013/0251706, 2013/0195846, 2013/0183290, 2013/0089540, 2013/0004480, 2012/0315268, 2012/0301459, 2012/0276085, 2012/0263713, 2012/0258102, 2012/0258101, 2012/0251534, 2012/0219549, 2012/0183545, 2012/0100133, 2012/0034185, 2011/0287006, 2011/0263825, 2011/0243931, 2011/0217298, 2011/0200598, 2011/0195022, 2011/0195021, 2011/0177067, 2011/0165159, 2011/0165152, 2011/0165151, 2011/0129412, 2011/0086025, 2011/0081681, 2011/0020322, 2010/0330089, 2010/0310581, 2010/0303808, 2010/0183601, 2010/0080769, 2009/0285795, 2009/0203886, 2009/0197330, 2009/0196879, 2009/0191195, 2009/0175854, 2009/0155253, 2009/0136516, 2009/0130089, 2009/0110688, 2009/0098118, 2009/0074760, 2009/0060913, 2009/0035322, 2008/0260641, 2008/0213273, 2008/0089885, 2008/0044421, 2008/0038261, 2007/0280882, 2007/0231324, 2007/0224189, 2007/0059306, 2007/0020259, 2007/0014785, 2007/0014720, 2006/0121032, 2005/0180972, 2005/0112060, 2005/0069545, 2005/0025764, 2004/0213784, 2004/0167319, 2004/0093621, 2003/0219433, 2003/0206903, 2003/0180292, 2003/0026804, 2002/0039557, 2002/0012665, and 2001/0018041, each herein incorporated by reference in their entirety (e.g., sections describing CD20 inhibitors).

[0549] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0550] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0551] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0552] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR). Additional examples of CD20 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD20 InhibitorsPeptides and Fusion Proteins

[0553] In some embodiments, the CD20 inhibitor is an immunotoxin (e.g., MT-3724 (Hamlin Blood 128: 4200, 2016).

[0554] In some embodiments, the CD20 inhibitor is a fusion protein (e.g., TRU-015 (Rubbert-Roth Curr. Opin. Mol. Ther 12(1): 115-123, 2010). Additional examples of CD20 inhibitors that are fusion proteins are described in, e.g., U.S. Patent Application Publication Nos. 2012/0195895, 2012/0034185, 2009/0155253, 2007/0020259, and 2003/0219433, each of which are herein incorporated by reference in their entirety (e.g., sections describing CD20 inhibitors).

CD25 Inhibitors

[0555] The term CD25 inhibitors refers to an agent which decreases the ability of CD25 (also called interleukin-2 receptor alpha chain) to bind to interleukin-2. CD25 forms a complex with interleukin-2 receptor beta chain and interleukin-2 common gamma chain.

[0556] In some embodiments, the CD25 inhibitor is an antibody or an antigen-binding fragment thereof, or a fusion protein. Exemplary CD25 inhibitors are described herein. Additional examples of CD25 inhibitors are known in the art.

[0557] An exemplary sequence of human CD25 is shown below.

TABLE-US-00008 HumanCD25Isoform1 (SEQIDNO:70) elcdddppeiphatfkamaykegtmlnceckrgfrriks gslymlctgnsshsswdnqcqctssatmttkqvtpqpee qkerkttemqspmqpvdqaslpghcrepppweneateriy hfvvgqmvyyqcvqgyralhrgpaesvckmthgktrwtqp qlictgemetsqfpgeekpqaspegrpesetsclvtadf qiqtemaatmetsiftteyqvavagcvfllisylllsglt wqrrqrksrrti HumanCD25Isoform2 (SEQIDNO:71) elcdddppeiphatfkamaykegtmlnceckrgfrriks gslymlctgnsshsswdnqcqctssatmttkqvtpqpee qkerkttemqspmqpvdqaslpgeekpqaspegrpesets clvtadfqiqtemaatmetsiftteyqvavagcvfllis vlllsgltwqrrqrksrrti HumanCD25Isoform3 (SEQIDNO:72) elcdddppeiphatfkamaykegtmlnceckrgfrriks gslymlctgnsshsswdnqcqctssatmttkqvtpqpee qkerkttemqspmqpvdqaslpdfqiqtemaatmetsift teyqvavagcvfflisvlllsgltwqrrqrksrrti

CD25 InhibitorsAntibodies

[0558] In some embodiments, the CD25 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, a CD25 inhibitor is an antibody or an antigen-binding fragment thereof that specifically binds to CD25. In some embodiments, a CD25 inhibitor is an antibody that specifically binds to IL-2.

[0559] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0560] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0561] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0562] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0563] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of basiliximab (Simulect) (Wang et al., Clin. Exp. Immunol. 155(3): 496-503, 2009; and Kircher et al., Clin. Exp. Immunol. 134(3): 426-430, 2003); daclizumab (Zenapax; Zinbryta) (Berkowitz et al., Clin. Immunol. 155(2): 176-187, 2014; and Bielekova et al., Arch Neurol. 66(4): 483-489, 2009); or IMTOX-25.

[0564] In some embodiments, the CD25 inhibitor is an antibody-drug-conjugate (e.g., ADCT-301 (Flynn et al., Blood 124: 4491, 2014)).

[0565] Additional examples of CD25 inhibitors that are antibodies are known in the art (see, e.g., WO 2004/045512). Additional examples of CD25 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos. 2017/0240640, 2017/0233481, 2015/0259424, 2015/0010539, 2015/0010538, 2012/0244069, 2009/0081219, 2009/0041775, 2008/0286281, 2008/0171017, 2004/0170626, 2001/0041179, and 2010/0055098, each of which is incorporated herein by reference (e.g., sections that describe CD25 inhibitors).

[0566] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0567] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0568] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0569] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M-is-1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.5106 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR). Additional examples of CD25 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD25 InhibitorsFusion Proteins

[0570] In some embodiments, the CD25 inhibitor is a fusion protein. See, e.g., Zhang et al., PNAS 100(4): 1891-1895, 2003.

CD28 Inhibitors

[0571] The term CD28 inhibitors refers to an agent which decreases the ability of CD28 to bind to one or both of CD80 and CD86. CD28 is a receptor that binds to its ligands, CD80 (also called B7.1) and CD86 (called B7.2).

[0572] In some embodiments, the CD28 inhibitor can decrease the binding between CD28 and CD80 by blocking the ability of CD28 to interact with CD80. In some embodiments, the CD28 inhibitor can decrease the binding between CD28 and CD86 by blocking the ability of CD28 to interact with CD86. In some embodiments, the CD28 inhibitor can decrease the binding of CD28 to each of CD80 and CD86.

[0573] In some embodiments, the CD28 inhibitor is an antibody or an antigen-binding fragment thereof, a fusion protein, or peptide. Exemplary CD28 inhibitors are described herein. Additional examples of CD28 inhibitors are known in the art.

[0574] Exemplary sequences for human CD28, human CD80, and human CD86 are shown below.

TABLE-US-00009 HumanCD28Isoform1 (SEQIDNO:73) nkilvkqspmlvaydnavnlsckysynlfsrefraslhkgld savevcvvygnysqqlqvysktgfncdgklgnesvtfylq nlyvnqtdiyfckievmypppyldneksngtiihvkgkhl cpsplfpgpskpfwvlvvvggvlacysllvtvafiifwvr skrsrllhsdymnmtprrpgptrkhyqpyapprdfaayrs HumanCD28Isoform2 (SEQIDNO:74) nkilvkqspmlvaydnavnlswkhlcpsplfpgpskpfwvlv vvggvlacysllvtvafiifwvrskrsrllhsdymnmtpr rpgptrkhyqpyapprdfaayrs HumanCD28Isoform3 (SEQIDNO:75) khlcpsplfpgpskpfwvlvvvggvlacysllvtvafiifwv rskrsrllhsdymnmtprrpgptrkhyqpyapprdfaayrs HumanCD80 (SEQIDNO:76) vihvtkevkevatlscghnvsveelaqtriywqkek kmvltmmsgdmniwpeyknrtifditnnlsivilalrpsd egtyecwlkyekdafkrehlaevtlsvkadfptpsisdf eiptsnirriicstsggfpephlswlengeelnainttvs qdpetelyavsskldfnmttnhsfinclikyghlrvnqtfn wnttkqehfpdnllpswaitlisvngifviccltycfapr crerrrnerlrresvrpv HumanCD86Isoform1 (SEQIDNO:77) yfnetadlpcqfansqnqslselvvfwqdqenlvinevyl gkekfdsvhskymgrtsfdsdswfirlhnlqikdkglyqc iihhkkptgmirihqmnselsvlanfsqpeivpisniten vyinitcssihgypepkkmsvllrtknstieydgimqksq dnvtelydvsislsysfpdvtsnmtifciletdktrllss pfsieledpqpppdhipwitavlptviicvmvfclilwkw kkkkrprnsykcgtntmereeseqtkkrekihipersdea qrvfkssktsscdksdtcf HumanCD86Isoform2 (SEQIDNO:78) yfnetadlpcqfansqnqslselvvfwqdgenlvin evylgkekfdsvhskymgrtsfdsdswtlrlhnlqikdkg lyqciihhkkptgmirihqmnselsvlanfsqpeivpisn itenvyinitcssihgypepkkmsyllrtknstieydgim qksqdnvtelydvsislsysfpdvtsnmtifciletdktr llsspfsieledpqpppdhipwitavlptviicvmvfcli lwkwkkkkrprnsykcgtntmereeseqtkkrekihiper sdeaqrvfkssktsscdksdtcf HumanCD86Isoform3 (SEQIDNO:79) yfnetadlpcqfansqnqslselvvfwqdgenlvin evylgkekfdsvhskymgrtsfdsdswtlrlhnlqikdkg lyqciihhkkptgmirihqmnselsvlanfsqpeivpisn itenvyinitcssihgypepkkmsyllrtknstieydgim qksqdnvtelydvsislsysfpdvtsnmtifciletdktr llsspfsigtntmereeseqtkkrekihipersdeaqrvf kssktsscdksdtcf HumanCD86Isoform4 (SEQIDNO:80) eivpisnitenvyinitcssihgypepkkmsvllrtknstiey dgimqksqdnvtelydvsislsysfpdvtsnmtifcilet dktrllsspfsieledpqpppdhipwitavlptviicvmv fclilwkwkkkkrprnsykcgtntmereeseqtkkrekih ipersdeaqrvfkssktsscdksdtcf HumanCD86Isoform5 (SEQIDNO:81) mgrtsfdsdswtlrlhnlqikdkglyqciihhkkptgmir ihqmnselsvlanfsqpeivpisnitenvyinitcssihg ypepkkmsvllrtknstieydgimqksqdnvtelydvsis lsysfpdvtsnmtifciletdktrllsspfsieledpqpp pdhipwitavlptviicvmvfclilwkwkkkkrprnsykc gtntmereeseqtkkrekihipersdeaqrvfkssktssc dksdtcf

CD28 InhibitorsAntibodies

[0575] In some embodiments, the CD28 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).

[0576] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0577] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0578] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0579] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0580] In some embodiments, the CD28 inhibitor is a monovalent Fab antibody (e.g., CFR104) (Poirier et al., Am. J. Transplant 15(1): 88-100, 2015).

[0581] Additional examples of CD28 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos. 2017/0240636, 2017/0114136, 2016/0017039, 2015/0376278, 2015/0299321, 2015/0232558, 2015/0150968, 2015/0071916, 2013/0266577, 2013/0230540, 2013/0109846, 2013/0078257, 2013/0078236, 2013/0058933, 2012/0201814, 2011/0097339, 2011/0059071, 2011/0009602, 2010/0266605, 2010/0028354, 2009/0246204, 2009/0117135, 2009/0117108, 2008/0095774, 2008/0038273, 2007/0154468, 2007/0134240, 2007/0122410, 2006/0188493, 2006/0165690, 2006/0039909, 2006/0009382, 2006/0008457, 2004/0116675, 2004/0092718, 2003/0170232, 2003/0086932, 2002/0006403, 2013/0197202, 2007/0065436, 2003/0180290, 2017/0015747, 2012/0100139, and 2007/0148162, each of which is incorporated by reference in its entirety (e.g., sections that described CD28 inhibitors).

[0582] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0583] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0584] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0585] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0586] Additional examples of CD28 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD28 InhibitorsFusion Proteins and Peptides

[0587] In some embodiments, the CD28 inhibitor is a fusion protein (see, e.g., U.S. Pat. No. 5,521,288; and US 2002/0018783). In some embodiments, the CD28 inhibitor is abatacept (Orencia) (Herrero-Beaumont et al., Rheumatol. Clin. 8: 78-83, 2012; and Korhonen and Moilanen Basic Clin. Pharmacol. Toxicol. 104(4): 276-284, 2009).

[0588] In some embodiments, the CD28 inhibitor is a peptide mimetic (e.g., AB103) (see, e.g., Bulger et al., JAMA Surg. 149(6): 528-536, 2014), or a synthetical peptoid (see, e.g., Li et al., Cell Mol. Immunol. 7(2): 133-142, 2010).

CD49 Inhibitors

[0589] The term CD49 inhibitors refers to an agent which decreases the ability of CD49 to bind to one of its ligands (e.g., MMP1). In some embodiments, the CD49 inhibitor is an antibody or an antigen-binding fragment thereof. Exemplary CD49 inhibitors are described herein. Additional examples of CD49 inhibitors are known in the art.

[0590] Exemplary sequences for human CD49 and human MMP1 are shown below.

TABLE-US-00010 HumanCD49 (SEQIDNO:82) mgpertgaaplplllvlalsqgilncclaynvglpeakif sgpsseqfgyavqqfinpkgnwllvgspwsgfpenrmgdv ykcpvdlstatceklnlqtstsipnvtemktnmslglilt rnmgtggfltcgplwaqqcgnqyyttgvcsdispdfqlsa sfspatqpcpslidvvvvcdesnsiypwdavknflekfvq gldigptktqvgliqyannprvvfnlntyktkeemivats qtsqyggdltntfgaiqyarkyaysaasggrrsatkvmvy vtdgeshdgsmlkavidqcnhdnilrfgiavlgylnrnal dtknlikeikaiasipteryffnvsdeaallekagtlgeq ifsiegtvqggdnfqmemsqvgfsadyssqndilmlgavg afgwsgtivqktshghlifpkqafdqilqdrnhssylgys vaaistgesthfvagapranytgqivlysvnengnitviq ahrgdqigsyfgsylcsydydkdfitdyllvgapmymsdl kkeegrvylftikkgilgqhqflegpegientrfgsaiaa lsdinmdgfndvivgsplenqnsgavyiynghqgfirtky sqkilgsdgafrshlqyfgrsldgygdlngdsitdvsiga fgqvvqlwsqsiadvaieasftpekitivnknaqiilklc fsakfrptkqnnqvaivynitldadgfssrvtsrglfken nerclqknmvvnqaqscpehiiyiqepsdvvnsldlrydi slenpgtspaleaysetakvfsipfhkdcgedglcisdlv ldvrqipaaqeqpfivsnqnkrltfsvtlknkresayntg ivvdfsenlffasfslpvdgtevtcqvaasqksvacdvgy palkreqqvtftinfdfnlqnlqnqaslsfqalsesqeen kadnlvnlkipllydaeihltrstninfyeissdgnvpsi vhsfedvgpkfifslkvttgsvpvsmatviihipqytkek nplmyltgvqtdkagdiscnadinplkigqtsssysfkse nfrhtkelncrtascsnvtcwlkdvhmkgeyfvnyttriw ngtfasstfqtvqltaaaeintynpeiyviedntytiplm imkpdekaevptgviigsiiagillllalvailwklgffk rkyekmtknpdeidettelss HumanMMP1 (SEQIDNO:83) mhsfpplllllfwgvvshsfpatletqeqdvdlyqkylek yynlkndgrqvekrrnsgpvveklkqmqeffglkvtgkpd aetlkvmkqprcgvpdvaqfvltegnprweqthltyrien ytpdlpradvdhaiekafqlwsnvtpltftkvsegqadim isfvrgdhrdnspfdgpggnlahafqpgpgiggdahfded erwtnnfreynlhrvaahelghslglshstdigalmypsy tfsgdyglaqddidgiqaiygrsqnpvqpigpqtpkacds kltfdaittirgevmffkdrfymrtnpfypevelnfisvf wpqlpngleaayefadrdevrffkgnkywavqgqnvlhgy pkdiyssfgfprtvkhidaalseentgktyffvankywry deykrsmdpgypkmiandfpgighkvdavfmkdgffyffh gtrqykfdpktkriltlqkanswfncrkn

CD49 InhibitorsAntibodies

[0591] In some embodiments, the CD49 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).

[0592] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0593] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0594] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0595] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0596] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of natalizumab (Tysabri; Antegren) (see, e.g., Pagnini et al., Expert Opin. Biol. Ther. 17(11): 1433-1438, 2017; and Chataway and Miller Neurotherapeutics 10(1): 19-28, 2013; or vatelizumab (ELND-004)).

[0597] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0598] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0599] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0600] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.5106 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR). Additional examples of CD49 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD89 Inhibitors

[0601] The term CD89 inhibitors refers to an agent which decreases the ability of CD89 to bind to IgA. CD89 is a transmembrane glycoprotein that binds to the heavy-chain constant region of IgA. In some embodiments, the CD89 inhibitor can decrease the binding between CD89 and IgA by blocking the ability of CD89 to interact with IgA. In some embodiments, the CD89 inhibitor is an antibody or an antigen-binding fragment thereof. Exemplary CD89 inhibitors are described herein. Additional examples of CD89 inhibitors are known in the art.

[0602] An exemplary sequence for human CD89 is shown below.

TABLE-US-00011 HumanCD89 (SEQIDNO:84) mdpkqttllclvlclgqriqaqegdfpmpfisaksspvip ldgsvkiqcqaireayltqlmiiknstyreigrrlkfwne tdpefvidhmdankagryqcqyrighyrfrysdtlelvvt glygkpflsadrglvlmpgenisltcssahipfdrfslak egelslpqhqsgehpanfslgpvdlnvsgiyrcygwynrs pylwsfpsnalelvvtdsihqdyttqnlirmavaglvlva llailvenwhshtalnkeasadvaepswsqqmcqpgltfa rtpsvck

CD89 InhibitorsAntibodies

[0603] In some embodiments, the CD89 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).

[0604] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al., Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0605] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0606] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0607] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0608] In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of HF-1020. Additional examples of CD89 antibodies are known in the art (see, e.g., WO 2002/064634).

[0609] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0610] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0611] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0612] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0613] Additional examples of CD89 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD283 (TLR3) Antibodies

[0614] In some embodiments, the therapeutic agent is PRV-300, for example, as described in PCT publication WO 2006/060513 which is incorporated by reference herein in its entirety. PRV-300 is an anti-Toll-Like Receptor 3 (TLR3)/CD283 monoclonal antibody that blocks TLR3 on cell surfaces and in endosomes.

IL-1 Inhibitors

[0615] The term IL-1 inhibitor refers to an agent that decreases the expression of an IL-1 cytokine or an IL-1 receptor and/or decreases the ability of an IL-1 cytokine to bind specifically to an IL-1 receptor. Non-limiting examples of IL-1 cytokines include IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, and IL-33. In some examples, an IL-1 cytokine is IL-1. In some examples, an IL-1 cytokine is IL-1.

[0616] As is known in the art, IL-1 and IL-1 each binds to a complex of IL-1R1 and IL1RAP proteins; IL-18 binds to IL-18R; IL-36, IL-36, and IL-36 each binds to a complex of IL-1RL2 and IL-1RAP proteins; and IL-33 binds to a complex of IL1RL1 and IL1RAP proteins. IL-1R is an endogenous soluble protein that decreases the ability of IL-la and IL-1 to bind to their receptor (e.g., a complex of IL-1R1 and IL1RAP proteins). IL-36R is an endogenous soluble protein that decreases the ability of IL-36, IL-36, and IL-36 to bind to their receptor (e.g., a complex of IL-1RL2 and IL-1RAP proteins).

[0617] In some embodiments, the IL-1 inhibitor mimicks native human interleukin 1 receptor antagonist (IL1-Ra).

[0618] In some embodiments, the IL-1 inhibitor targets IL-1. In some embodiments, the IL-1 inhibitor targets IL-1. In some embodiments, the IL-1 inhibitor targets one or both of IL-1R1 and IL1RAP. For example, an IL-1 inhibitor can decrease the expression of IL-1 and/or decrease the ability of IL-1 to bind to its receptor (e.g., a complex of IL-1R1 and IL1RAP proteins). In another example, an IL-1 inhibitor can decrease the expression of IL-1 and/or decrease the ability of IL-1 to binds to its receptor (e.g., a complex of IL-1R1 and IL1RAP proteins). In some embodiments, an IL-1 inhibitor can decrease the expression of one or both of IL-1R1 and IL1RAP.

[0619] In some embodiments, the IL-1 inhibitor targets IL-18. In some embodiments, the IL-1 inhibitor targets IL-18R. In some embodiments, the IL-1 inhibitor decreases the ability of IL-18 to bind to its receptor (e.g., IL-18Ra). In some embodiments, the IL-1 inhibitor decreases the expression of IL-18. In some embodiments, the IL-1 inhibitor decreases the expression of IL-18Ra.

[0620] In some embodiments, the IL-1 inhibitor targets one or more (e.g., two or three) of IL-36, IL-36, and IL-36. In some embodiments, the IL-1 inhibitor targets one or both of IL-1RL2 and IL-1RAP. In some embodiments, the IL-1 inhibitor decreases the expression of one or more (e.g., two or three) of IL-36, IL-36, and IL-36. In some embodiments, the IL-1 inhibitor decreases the expression of one or both of IL-1RL2 and IL-1RAP proteins. In some embodiments, the IL-1 inhibitor decreases the ability of IL-36a to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP). In some examples, the IL-1 inhibitor decreases the ability of IL-36 to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP). In some examples, the IL-1 inhibitor decreases the ability of IL-36 to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP).

[0621] In some embodiments, the IL-1 inhibitor targets IL-33. In some embodiments, the IL-1 inhibitor targets one or both of IL1RL1 and IL1RAP. In some embodiments, the IL-1 inhibitor decreases the expression of IL-33. In some embodiments, the IL-1 inhibitor decreases the expression of one or both of IL1RL1 and IL1RAP. In some embodiments, the IL-1 inhibitor decreases the ability of IL-33 to bind to its receptor (e.g., a complex of IL1RL1 and IL1RAP proteins).

[0622] In some embodiments, an IL-1 inhibitory agent is an inhibitory nucleic acid, an antibody or fragment thereof, or a fusion protein. In some embodiments, the inhibitory nucleic acid is an antisense nucleic acid, a ribozyme, or a small interfering RNA.

Inhibitory Nucleic Acids

[0623] Inhibitory nucleic acids that can decrease the expression of IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 85-125).

TABLE-US-00012 HumanIL-1mRNA (SEQIDNO:85) 1 agtaaccaggcaacaccattgaaggctcatatgtaaaaatccatgccttcctttctccca 61 atctccattcccaaacttagccactggcttctggctgaggccttacgcatacctcccggg 121 gcttgcacacaccttcttctacagaagacacaccttgggcatatcctacagaagaccagg 181 cttctctctggtccttggtagagggctactttactgtaacagggccagggtggagagttc 241 tctcctgaagctccatcccctctataggaaatgtgttgacaatattcagaagagtaagag 301 gatcaagacttctttgtgctcaaataccactgttctcttctctaccctgccctaaccagg 361 agcttgtcaccccaaactctgaggtgatttatgccttaatcaagcaaacttccctcttca 421 gaaaagatggctcattttccctcaaaagttgccaggagctgccaagtattctgccaattc 481 accctggagcacaatcaacaaattcagccagaacacaactacagctactattagaactat 541 tattattaataaattcctctccaaatctagccccttgacttcggatttcacgatttctcc 601 cttcctcctagaaacttgataagtttcccgcgcttccctttttctaagactacatgtttg 661 tcatcttataaagcaaaggggtgaataaatgaaccaaatcaataacttctggaatatctg 721 caaacaacaataatatcagctatgccatctttcactattttagccagtatcgagttgaat 781 gaacatagaaaaatacaaaactgaattcttccctgtaaattccccgttttgacgacgcac 841 ttgtagccacgtagccacgcctacttaagacaattacaaaaggcgaagaagactgactca 901 ggcttaagctgccagccagagagggagtcatttcattggcgtttgagtcagcaaagaagt 961 caagatggccaaagttccagacatgtttgaagacctgaagaactgttacagtgaaaatga 1021 agaagacagttcctccattgatcatctgtctctgaatcagaaatccttctatcatgtaag 1081 ctatggcccactccatgaaggctgcatggatcaatctgtgtctctgagtatctctgaaac 1141 ctctaaaacatccaagcttaccttcaaggagagcatggtggtagtagcaaccaacgggaa 1201 ggttctgaagaagagacggttgagtttaagccaatccatcactgatgatgacctggaggc 1261 catcgccaatgactcagaggaagaaatcatcaagcctaggtcagcaccttttagcttcct 1321 gagcaatgtgaaatacaactttatgaggatcatcaaatacgaattcatcctgaatgacgc 1381 cctcaatcaaagtataattcgagccaatgatcagtacctcacggctgctgcattacataa 1441 tctggatgaagcagtgaaatttgacatgggtgcttataagtcatcaaaggatgatgctaa 1501 aattaccgtgattctaagaatctcaaaaactcaattgtatgtgactgcccaagatgaaga 1561 ccaaccagtgctgctgaaggagatgcctgagatacccaaaaccatcacaggtagtgagac 1621 caacctcctcttcttctgggaaactcacggcactaagaactatttcacatcagttgccca 1681 tccaaacttgtttattgccacaaagcaagactactgggtgtgcttggcaggggggccacc 1741 ctctatcactgactttcagatactggaaaaccaggcgtaggtctggagtctcacttgtct 1801 cacttgtgcagtgttgacagttcatatgtaccatgtacatgaagaagctaaatcctttac 1861 tgttagtcatttgctgagcatgtactgagccttgtaattctaaatgaatgtttacactct 1921 ttgtaagagtggaaccaacactaacatataatgttgttatttaaagaacaccctatattt 1981 tgcatagtaccaatcattttaattattattcttcataacaattttaggaggaccagagct 2041 actgactatggctaccaaaaagactctacccatattacagatgggcaaattaaggcataa 2101 gaaaactaagaaatatgcacaatagcagttgaaacaagaagccacagacctaggatttca 2161 tgatttcatttcaactgtttgccttctacttttaagttgctgatgaactcttaatcaaat 2221 agcataagtttctgggacctcagttttatcattttcaaaatggagggaataatacctaag 2281 ccttcctgccgcaacagttttttatgctaatcagggaggtcattttggtaaaatacttct 2341 tgaagccgagcctcaagatgaaggcaaagcacgaaatgttattttttaattattatttat 2401 atatgtatttataaatatatttaagataattataatatactatatttatgggaacccctt 2461 catcctctgagtgtgaccaggcatcctccacaatagcagacagtgttttctgggataagt 2521 aagtttgatttcattaatacagggcattttggtccaagttgtgcttatcccatagccagg 2581 aaactctgcattctagtacttgggagacctgtaatcatataataaatgtacattaattac 2641 cttgagccagtaattggtccgatctttgactcttttgccattaaacttacctgggcattc 2701 ttgtttcaattccacctgcaatcaagtcctacaagctaaaattagatgaactcaactttg 2761 acaaccatgagaccactgttatcaaaactttcttttctggaatgtaatcaatgtttcttc 2821 taggttctaaaaattgtgatcagaccataatgttacattattatcaacaatagtgattga 2881 tagagtgttatcagtcataactaaataaagcttgcaacaaaattctctgacaaaaaaaaa 2941 aaaaaaa HumanIL-1mRNA (SEQIDNO:86) 1 accaaacctcttcgaggcacaaggcacaacaggctgctctgggattctcttcagccaatc 61 ttcattgctcaagtgtctgaagcagccatggcagaagtacctgagctcgccagtgaaatg 121 atggcttattacagtggcaatgaggatgacttgttctttgaagctgatggccctaaacag 181 atgaagtgctccttccaggacctggacctctgccctctggatggcggcatccagctacga 241 atctccgaccaccactacagcaagggcttcaggcaggccgcgtcagttgttgtggccatg 301 gacaagctgaggaagatgctggttccctgcccacagaccttccaggagaatgacctgagc 361 accttctttcccttcatctttgaagaagaacctatcttcttcgacacatgggataacgag 421 gcttatgtgcacgatgcacctgtacgatcactgaactgcacgctccgggactcacagcaa 481 aaaagcttggtgatgtctggtccatatgaactgaaagctctccacctccagggacaggat 541 atggagcaacaagtggtgttctccatgtcctttgtacaaggagaagaaagtaatgacaaa 601 atacctgtggccttgggcctcaaggaaaagaatctgtacctgtcctgcgtgttgaaagat 661 gataagcccactctacagctggagagtgtagatcccaaaaattacccaaagaagaagatg 721 gaaaagcgatttgtcttcaacaagatagaaatcaataacaagctggaatttgagtctgcc 781 cagttccccaactggtacatcagcacctctcaagcagaaaacatgcccgtcttcctggga 841 gggaccaaaggcggccaggatataactgacttcaccatgcaatttgtgtcttcctaaaga 901 gagctgtacccagagagtcctgtgctgaatgtggactcaatccctagggctggcagaaag 961 ggaacagaaaggtttttgagtacggctatagcctggactttcctgttgtctacaccaatg 1021 cccaactgcctgccttagggtagtgctaagaggatctcctgtccatcagccaggacagtc 1081 agctctctcctttcagggccaatccccagcccttttgttgagccaggcctctctcacctc 1141 tcctactcacttaaagcccgcctgacagaaaccacggccacatttggttctaagaaaccc 1201 tctgtcattcgctcccacattctgatgagcaaccgcttccctatttatttatttatttgt 1261 ttgtttgttttattcattggtctaatttattcaaagggggcaagaagtagcagtgtctgt 1321 aaaagagcctagtttttaatagctatggaatcaattcaatttggactggtgtgctctctt 1381 taaatcaagtcctttaattaagactgaaaatatataagctcagattatttaaatgggaat 1441 atttataaatgagcaaatatcatactgttcaatggttctgaaataaacttcactgaag HumanIL-18mRNAVariant1 (SEQIDNO:87) 1 attctctccccagcttgctgagccctttgctcccctggcgactgcctggacagtcagcaa 61 ggaattgtctcccagtgcattttgccctcctggctgccaactctggctgctaaagcggct 121 gccacctgctgcagtctacacagcttcgggaagaggaaaggaacctcagaccttccagat 181 cgcttcctctcgcaacaaactatttgtcgcaggaataaagatggctgctgaaccagtaga 241 agacaattgcatcaactttgtggcaatgaaatttattgacaatacgctttactttatagc 301 tgaagatgatgaaaacctggaatcagattactttggcaagcttgaatctaaattatcagt 361 cataagaaatttgaatgaccaagttctcttcattgaccaaggaaatcggcctctatttga 421 agatatgactgattctgactgtagagataatgcaccccggaccatatttattataagtat 481 gtataaagatagccagcctagaggtatggctgtaactatctctgtgaagtgtgagaaaat 541 ttcaactctctcctgtgagaacaaaattatttcctttaaggaaatgaatcctcctgataa 601 catcaaggatacaaaaagtgacatcatattctttcagagaagtgtcccaggacatgataa 661 taagatgcaatttgaatcttcatcatacgaaggatactttctagcttgtgaaaaagagag 721 agacctttttaaactcattttgaaaaaagaggatgaattgggggatagatctataatgtt 781 cactgttcaaaacgaagactagctattaaaatttcatgccgggcgcagtggctcacgcct 841 gtaatcccagccctttgggaggctgaggcgggcagatcaccagaggtcaggtgttcaaga 901 ccagcctgaccaacatggtgaaacctcatctctactaaaaatacaaaaaattagctgagt 961 gtagtgacgcatgccctcaatcccagctactcaagaggctgaggcaggagaatcacttgc 1021 actccggaggtagaggttgtggtgagccgagattgcaccattgcgctctagcctgggcaa 1081 caacagcaaaactccatctcaaaaaataaaataaataaataaacaaataaaaaattcata 1141 atgtgaaaaaaaaaaaaaaaaaa HumanIL-18mRNAVariant2 (SEQIDNO:88) 1 attctctccccagcttgctgagccctttgctcccctggcgactgcctggacagtcagcaa 61 ggaattgtctcccagtgcattttgccctcctggctgccaactctggctgctaaagcggct 121 gccacctgctgcagtctacacagcttcgggaagaggaaaggaacctcagaccttccagat 181 cgcttcctctcgcaacaaactatttgtcgcaggaataaagatggctgctgaaccagtaga 241 agacaattgcatcaactttgtggcaatgaaatttattgacaatacgctttactttataga 301 aaacctggaatcagattactttggcaagcttgaatctaaattatcagtcataagaaattt 361 gaatgaccaagttctcttcattgaccaaggaaatcggcctctatttgaagatatgactga 421 ttctgactgtagagataatgcaccccggaccatatttattataagtatgtataaagatag 481 ccagcctagaggtatggctgtaactatctctgtgaagtgtgagaaaatttcaactctctc 541 ctgtgagaacaaaattatttcctttaaggaaatgaatcctcctgataacatcaaggatac 601 aaaaagtgacatcatattctttcagagaagtgtcccaggacatgataataagatgcaatt 661 tgaatcttcatcatacgaaggatactttctagcttgtgaaaaagagagagacctttttaa 721 actcattttgaaaaaagaggatgaattgggggatagatctataatgttcactgttcaaaa 781 cgaagactagctattaaaatttcatgccgggcgcagtggctcacgcctgtaatcccagcc 841 ctttgggaggctgaggcgggcagatcaccagaggtcaggtgttcaagaccagcctgacca 901 acatggtgaaacctcatctctactaaaaatacaaaaaattagctgagtgtagtgacgcat 961 gccctcaatcccagctactcaagaggctgaggcaggagaatcacttgcactccggaggta 1021 gaggttgtggtgagccgagattgcaccattgcgctctagcctgggcaacaacagcaaaac 1081 tccatctcaaaaaataaaataaataaataaacaaataaaaaattcataatgtgaaaaaaa 1141 aaaaaaaaaaa HumanIL-36mRNA (SEQIDNO:89) 1 aaaacccaagtgcagtagaagccattgttcataatggtagggatacagggtccttcgtaa 61 cagattatcagtgtggcctatgctggaaagtctggtgacctctgattttttttgcttcca 121 ggtctttggccttggcactctttgtcatattagagttcctgggtctaggcctgggcagga 181 ttcataggtgcagctgcttctgctggaggtagactgcatccaacaaagtaagggtgctgg 241 gtgagttctgggagtatagattctgactggggtcactgctgggctggccgccagtctttc 301 atctgacccagggttaaactgtggcttgggactgactcaggtcctctcttggggtcggtc 361 tgcacataaaaggactcctatccttggcagttctgaaacaacaccaccacaatggaaaaa 421 gcattgaaaattgacacacctcagcaggggagcattcaggatatcaatcatcgggtgtgg 481 gttcttcaggaccagacgctcatagcagtcccgaggaaggaccgtatgtctccagtcact 541 attgccttaatctcatgccgacatgtggagacccttgagaaagacagagggaaccccatc 601 tacctgggcctgaatggactcaatctctgcctgatgtgtgctaaagtcggggaccagccc 661 acactgcagctgaaggaaaaggatataatggatttgtacaaccaacccgagcctgtgaag 721 tcctttctcttctaccacagccagagtggcaggaactccaccttcgagtctgtggctttc 781 cctggctggttcatcgctgtcagctctgaaggaggctgtcctctcatccttacccaagaa 841 ctggggaaagccaacactactgactttgggttaactatgctgttttaa HumanIL-36mRNAVariant1 (SEQIDNO:90) 1 cacgggttcctccccactctgtctttctcacctctccttcacttttcctagcctcctcac 61 caccatctgatctatcttgttctcttcacaaaaggctctgaagacatcatgaacccacaa 121 cgggaggcagcacccaaatcctatgctattcgtgattctcgacagatggtgtgggtcctg 181 agtggaaattctttaatagcagctcctcttagccgcagcattaagcctgtcactcttcat 241 ttaatagcctgtagagacacagaattcagtgacaaggaaaagggtaatatggtttacctg 301 ggaatcaagggaaaagatctctgtctcttctgtgcagaaattcagggcaagcctactttg 361 cagcttaagcttcagggctcccaagataacatagggaaggacacttgctggaaactagtt 421 ggaattcacacatgcataaacctggatgtgagagagagctgcttcatgggaacccttgac 481 caatggggaataggagtgggtagaaagaagtggaagagttcctttcaacatcaccatctc 541 aggaagaaggacaaagatttctcatccatgcggaccaacataggaatgccaggaaggatg 601 tagaaataaggggaggaagattcccatctctacaatctttgagtgggtttgctatcaatg 661 aaatgctacaaatggaataagttgcagaaatttttctcttttcttgggttctggagagtt 721 tgtaaaacaaggacactatgtatttttaaagagttggtaaatcttacctgtaaagctaga 781 gaaggtcggagtctttttaggagtagatttggactacataacctgtaaatgtgttttgtc 841 cagtccttagagtgttttttaaaaaattgtaaagtcaaggttttcatgaaaaatgggaag 901 atcagacaacattgctcctgaattcccacagagcagcaagctactagagctcaatctgtt 961 atttcttttcctgatgtacaggggttaagtcctatggaagaaacagcagaattattcaaa 1021 attatttacataatgtgcaattattcactagagcatgaggagtgaaacgctctgtttagt 1081 atgtataacttaaaaggaacacatacaattaaaagtaattgaaagacatttcttcttaaa 1141 aattctataatcttacactggtaaaataaactagtttttcccatgt HumanIL-36mRNAVariant2 (SEQIDNO:91) 1 cacgggttcctccccactctgtctttctcacctctccttcacttttcctagcctcctcac 61 caccatctgatctatcttgttctcttcacaaaaggctctgaagacatcatgaacccacaa 121 cgggaggcagcacccaaatcctatgctattcgtgattctcgacagatggtgtgggtcctg 181 agtggaaattctttaatagcagctcctcttagccgcagcattaagcctgtcactcttcat 241 ttaatagcctgtagagacacagaattcagtgacaaggaaaagggtaatatggtttacctg 301 ggaatcaagggaaaagatctctgtctcttctgtgcagaaattcagggcaagcctactttg 361 cagcttaaggaaaaaaatatcatggacctgtatgtggagaagaaagcacagaagcccttt 421 ctctttttccacaataaagaaggctccacttctgtctttcagtcagtctcttaccctggc 481 tggttcatagccacctccaccacatcaggacagcccatctttctcaccaaggagagaggc 541 ataactaataacactaacttctacttagattctgtggaataaatccagcctaggctgtgg 601 gtggctggttccaggatagagaatcaagctgtcagagtcatcttaacagatcattatgcg 661 actgagttcactagcagttcagcccatccatagcttacctcattcttactatccaaaagc 721 cacctcctcctccaaacatccatttctgtaccaagaccctcactcgaatgtcactatccc 781 aagatgaaacctaaaaatcactttccattctttcttgatcttaccccaccatccactcag 841 ctgccatgcccagtttagtcaaccccccaaatgctgcttcatgcaaccttccattcctat 901 tccttttgccaacccatgatgtagagatgtggattcatgacattttgttcatacaacttc 961 ttcaataaaacattataatatgtgccccaaagataaagctgaagaatgagatgaatgtga 1021 aattaaaggtttgcatgtctttctaatcctaaaaaaaaaaaaaaaaaa HumanIL-36mRNAVariant1 (SEQIDNO:92) 1 gaagctgctggagccacgattcagtcccctggactgtagataaagaccctttcttgccag 61 gtgctgagacaaccacactatgagaggcactccaggagacgctgatggtggaggaagggc 121 cgtctatcaatcaatgtgtaaacctattactgggactattaatgatttgaatcagcaagt 181 gtggacccttcagggtcagaaccttgtggcagttccacgaagtgacagtgtgaccccagt 241 cactgttgctgttatcacatgcaagtatccagaggctcttgagcaaggcagaggggatcc 301 catttatttgggaatccagaatccagaaatgtgtttgtattgtgagaaggttggagaaca 361 gcccacattgcagctaaaagagcagaagatcatggatctgtatggccaacccgagcccgt 421 gaaacccttccttttctaccgtgccaagactggtaggacctccacccttgagtctgtggc 481 cttcccggactggttcattgcctcctccaagagagaccagcccatcattctgacttcaga 541 acttgggaagtcatacaacactgcctttgaattaaatataaatgactgaactcagcctag 601 aggtggcagcttggtctttgtcttaaagtttctggttcccaatgtgttttcgtctacatt 661 ttcttagtgtcattttcacgctggtgctgagacaggggcaaggctgctgttatcatctca 721 ttttataatgaagaagaagcaattacttcatagcaactgaagaacaggatgtggcctcag 781 aagcaggagagctgggtggtataaggctgtcctctcaagctggtgctgtgtaggccacaa 841 ggcatctgcatgagtgactttaagactcaaagaccaaacactgagctttcttctaggggt 901 gggtatgaagatgcttcagagctcatgcgcgttacccacgatggcatgactagcacagag 961 ctgatctctgtttctgttttgctttattccctcttgggatgatatcatccagtctttata 1021 tgttgccaatatacctcattgtgtgtaatagaaccttcttagcattaagaccttgtaaac 1081 aaaaataattcttgtgttaagttaaatcatttttgtcctaattgtaatgtgtaatcttaa 1141 agttaaataaactttgtgtatttatataataataaagctaaaactgatataaaataaaga 1201 aagagtaaactg HumanIL-36mRNAVariant2 (SEQIDNO:93) 1 gaagctgctggagccacgattcagtcccctggactgtagataaagaccctttcttgccag 61 gtgctgagacaaccacactatgagaggcactccaggagacgctgatggtggaggaagggc 121 cgtctatcaatcaatcactgttgctgttatcacatgcaagtatccagaggctcttgagca 181 aggcagaggggatcccatttatttgggaatccagaatccagaaatgtgtttgtattgtga 241 gaaggttggagaacagcccacattgcagctaaaagagcagaagatcatggatctgtatgg 301 ccaacccgagcccgtgaaacccttccttttctaccgtgccaagactggtaggacctccac 361 ccttgagtctgtggccttcccggactggttcattgcctcctccaagagagaccagcccat 421 cattctgacttcagaacttgggaagtcatacaacactgcctttgaattaaatataaatga 481 ctgaactcagcctagaggtggcagcttggtctttgtcttaaagtttctggttcccaatgt 541 gttttcgtctacattttcttagtgtcattttcacgctggtgctgagacaggggcaaggct 601 gctgttatcatctcattttataatgaagaagaagcaattacttcatagcaactgaagaac 661 aggatgtggcctcagaagcaggagagctgggtggtataaggctgtcctctcaagctggtg 721 ctgtgtaggccacaaggcatctgcatgagtgactttaagactcaaagaccaaacactgag 781 ctttcttctaggggtgggtatgaagatgcttcagagctcatgcgcgttacccacgatggc 841 atgactagcacagagctgatctctgtttctgttttgctttattccctcttgggatgatat 901 catccagtctttatatgttgccaatatacctcattgtgtgtaatagaaccttcttagcat 961 taagaccttgtaaacaaaaataattcttgtgttaagttaaatcatttttgtcctaattgt 1021 aatgtgtaatcttaaagttaaataaactttgtgtatttatataataataaagctaaaact 1081 gatataaaataaagaaagagtaaactg HumanIL-38mRNAVariant1 (SEQIDNO:94) 1 ggcagtgggactgggtttgagctgggcttatcctccaactgtgagggaggctacagcaca 61 ctccaccccactctcagggctgggaattgttgtggctcagctatttgggggaatctgttt 121 tccagtttctcagaaccagcgcaagcacacacatcccaggctcacacccctggtggctgg 181 acttgctcccggatagcctcagtcagggagaggcagagctgcctggagcctgctgggctg 241 gtggaagccttggtggattctggcaggccaattatagacgaatggcctggggaacccgtg 301 cagcccttggctgagtggttctaagccccagcacgtctgcctctggcttcacccagcctc 361 cttttctaactgcccttctctcctccccatcagtgaggaccagacaccactgattgcagg 421 aatgtgttccctccccatggcaagatactacataattaaatatgcagaccagaaggctct 481 atacacaagagatggccagctgctggtgggagatcctgttgcagacaactgctgtgcaga 541 gaagatctgcatacttcctaacagaggcttggcccgcaccaaggtccccattttcctggg 601 gatccagggagggagccgctgcctggcatgtgtggagacagaagaggggccttccctaca 661 gctggaggatgtgaacattgaggaactgtacaaaggtggtgaagaggccacacgcttcac 721 cttcttccagagcagctcaggctccgccttcaggcttgaggctgctgcctggcctggctg 781 gttcctgtgtggcccggcagagccccagcagccagtacagctcaccaaggagagtgagcc 841 ctcagcccgtaccaagttttactttgaacagagctggtagggagacaggaaactgcgttt 901 tagccttgtgcccccaaaccaagctcatcctgctcagggtctatggtaggcagaataatg 961 tcccccgaaatatgtccacatcctaatcccaagatctgtgcatatgttaccatacatgtc 1021 caaagaggttttgcaaatgtgattatgttaaggatcttgaaatgaggagacaatcctggg 1081 ttatccttgtgggctcagtttaatcacaagaaggaggcaggaagggagagtcagagagag 1141 aatggaagataccatgcttctaattttgaagatggagtgaggggccttgagccaacaaat 1201 gcaggtgtttttagaaggtggaaaagccaagggaacggattctcctctagagtctccgga 1261 aggaacacagctcttgacacatggatttcagctcagtgacacccatttcagacttctgac 1321 ctccacaactataaaataataaacttgtgttattgtaaacctctaa HumanIL-38mRNAVariant2 (SEQIDNO:95) 1 agttggagtctccagggatcagggttccaggaactcaggatctgcagtgaggaccagaca 61 ccactgattgcaggaatgtgttccctccccatggcaagatactacataattaaatatgca 121 gaccagaaggctctatacacaagagatggccagctgctggtgggagatcctgttgcagac 181 aactgctgtgcagagaagatctgcatacttcctaacagaggcttggcccgcaccaaggtc 241 cccattttcctggggatccagggagggagccgctgcctggcatgtgtggagacagaagag 301 gggccttccctacagctggaggatgtgaacattgaggaactgtacaaaggtggtgaagag 361 gccacacgcttcaccttcttccagagcagctcaggctccgccttcaggcttgaggctgct 421 gcctggcctggctggttcctgtgtggcccggcagagccccagcagccagtacagctcacc 481 aaggagagtgagccctcagcccgtaccaagttttactttgaacagagctggtagggagac 541 aggaaactgcgttttagccttgtgcccccaaaccaagctcatcctgctcagggtctatgg 601 taggcagaataatgtcccccgaaatatgtccacatcctaatcccaagatctgtgcatatg 661 ttaccatacatgtccaaagaggttttgcaaatgtgattatgttaaggatcttgaaatgag 721 gagacaatcctgggttatccttgtgggctcagtttaatcacaagaaggaggcaggaaggg 781 agagtcagagagagaatggaagataccatgcttctaattttgaagatggagtgaggggcc 841 ttgagccaacaaatgcaggtgtttttagaaggtggaaaagccaagggaacggattctcct 901 ctagagtctccggaaggaacacagctcttgacacatggatttcagctcagtgacacccat 961 ttcagacttctgacctccacaactataaaataataaacttgtgttattgtaaacctctaa 1021 aaaaaaa HumanIL-33mRNAVariant1 (SEQIDNO:96) 1 agtctacagactcctccgaacacagagctgcagctcttcagggaagaaatcaaaacaaga 61 tcacaagaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccaca 121 gcaaagtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacag 181 aaggccaaagaagtttgccccatgtactttatgaagctccgctctggccttatgataaaa 241 aaggaggcctgttactttaggagagaaaccaccaaaaggccttcactgaaaacaggtaga 301 aagcacaaaagacatctggtactcgctgcctgtcaacagcagtctactgtggagtgcttt 361 gcctttggtatatcaggggtccagaaatatactagagcacttcatgattcaagtatcaca 421 ggaatttcacctattacagagtatcttgcttctctaagcacatacaatgatcaatccatt 481 acttttgctttggaggatgaaagttatgagatatatgttgaagacttgaaaaaagatgaa 541 aagaaagataaggtgttactgagttactatgagtctcaacacccctcaaatgaatcaggt 601 gacggtgttgatggtaagatgttaatggtaaccctgagtcctacaaaagacttctggttg 661 catgccaacaacaaggaacactctgtggagctccataagtgtgaaaaaccactgccagac 721 caggccttctttgtccttcataatatgcactccaactgtgtttcatttgaatgcaagact 781 gatcctggagtgtttataggtgtaaaggataatcatcttgctctgattaaagtagactct 841 tctgagaatttgtgtactgaaaatatcttgtttaagctctctgaaacttagttgatggaa 901 acctgtgagtcttgggttgagtacccaaatgctaccactggagaaggaatgagagataaa 961 gaaagagacaggtgacatctaagggaaatgaagagtgcttagcatgtgtggaatgttttc 1021 catattatgtataaaaatattttttctaatcctccagttattcttttatttccctctgta 1081 taactgcatcttcaatacaagtatcagtatattaaatagggtattggtaaagaaacggtc 1141 aacattctaaagagatacagtctgacctttacttttctctagtttcagtccagaaagaac 1201 ttcatatttagagctaaggccactgaggaaagagccatagcttaagtctctatgtagaca 1261 gggatccattttaaagagctacttagagaaataattttccacagttccaaacgataggct 1321 caaacactagagctgctagtaaaaagaagaccagatgcttcacagaattatcattttttc 1381 aactggaataaaacaccaggtttgtttgtagatgtcttaggcaacactcagagcagatct 1441 cccttactgtcaggggatatggaacttcaaaggcccacatggcaagccaggtaacataaa 1501 tgtgtgaaaaagtaaagataactaaaaaatttagaaaaataaatccagtatttgtaaagt 1561 gaataacttcatttctaattgtttaatttttaaaattctgatttttatatattgagttta 1621 agcaaggcattcttacacgaggaagtgaagtaaattttagttcagacataaaatttcact 1681 tattaggaatatgtaacatgctaaaacttttttttttttaaagagtactgagtcacaaca 1741 tgttttagagcatccaagtaccatataatccaactatcatggtaaggccagaaatcttct 1801 aacctaccagagcctagatgagacaccgaattaacattaaaatttcagtaactgactgtc 1861 cctcatgtccatggcctaccatcccttctgaccctggcttccagggacctatgtctttta 1921 atactcactgtcacattgggcaaagttgcttctaatccttatttcccatgtgcacaagtc 1981 tttttgtattccagcttcctgataacactgcttactgtggaatattcatttgacatctgt 2041 ctcttttcatttcttttaactaccatgcccttgatatatcttttgcacctgctgaacttc 2101 atttctgtatcacctgacctctggatgccaaaacgtttattctgctttgtctgttgtaga 2161 attttagataaagctattaatggcaatatttttttgctaaacgtttttgttttttactgt 2221 cactagggcaataaaatttatactcaaccatataataacattttttaactactaaaggag 2281 tagtttttattttaaagtcttagcaatttctattacaacttttcttagacttaacactta 2341 tgataaatgactaacatagtaacagaatctttatgaaatatgaccttttctgaaaataca 2401 tacttttacatttctactttattgagacctattagatgtaagtgctagtagaatataaga 2461 taaaagaggctgagaattaccatacaagggtattacaactgtaaaacaatttatctttgt 2521 ttcattgttctgtcaataattgttaccaaagagataaaaataaaagcagaatgtatatca 2581 tcccatctgaaaaacactaattattgacatgtgcatctgtacaataaacttaaaatgatt 2641 attaaataatcaaatatatctactacattgtttatattattgaataaagtatattttcca 2701 aatgtaaaaaaaaaaaaa HumanIL-33mRNAVariant2 (SEQIDNO:97) 1 agtctacagactcctccgaacacagagctgcagctcttcagggaagaaatcaaaacaaga 61 tcacaagaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccaca 121 gcaaagtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacag 181 aaggccaaagaagtttgccccatgtactttatgaagctccgctctggccttatgataaaa 241 aaggaggcctgttactttaggagagaaaccaccaaaaggccttcactgaaaacaggtaga 301 aagcacaaaagacatctggtactcgctgcctgtcaacagcagtctactgtggagtgcttt 361 gcctttggtatatcaggggtccagaaatatactagagcacttcatgattcaagtatcaca 421 gataaggtgttactgagttactatgagtctcaacacccctcaaatgaatcaggtgacggt 481 gttgatggtaagatgttaatggtaaccctgagtcctacaaaagacttctggttgcatgcc 541 aacaacaaggaacactctgtggagctccataagtgtgaaaaaccactgccagaccaggcc 601 ttctttgtccttcataatatgcactccaactgtgtttcatttgaatgcaagactgatcct 661 ggagtgtttataggtgtaaaggataatcatcttgctctgattaaagtagactcttctgag 721 aatttgtgtactgaaaatatcttgtttaagctctctgaaacttagttgatggaaacctgt 781 gagtcttgggttgagtacccaaatgctaccactggagaaggaatgagagataaagaaaga 841 gacaggtgacatctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatatt 901 atgtataaaaatattttttctaatcctccagttattcttttatttccctctgtataactg 961 catcttcaatacaagtatcagtatattaaatagggtattggtaaagaaacggtcaacatt 1021 ctaaagagatacagtctgacctttacttttctctagtttcagtccagaaagaacttcata 1081 tttagagctaaggccactgaggaaagagccatagcttaagtctctatgtagacagggatc 1141 cattttaaagagctacttagagaaataattttccacagttccaaacgataggctcaaaca 1201 ctagagctgctagtaaaaagaagaccagatgcttcacagaattatcattttttcaactgg 1261 aataaaacaccaggtttgtttgtagatgtcttaggcaacactcagagcagatctccctta 1321 ctgtcaggggatatggaacttcaaaggcccacatggcaagccaggtaacataaatgtgtg 1381 aaaaagtaaagataactaaaaaatttagaaaaataaatccagtatttgtaaagtgaataa 1441 cttcatttctaattgtttaatttttaaaattctgatttttatatattgagtttaagcaag 1501 gcattcttacacgaggaagtgaagtaaattttagttcagacataaaatttcacttattag 1561 gaatatgtaacatgctaaaacttttttttttttaaagagtactgagtcacaacatgtttt 1621 agagcatccaagtaccatataatccaactatcatggtaaggccagaaatcttctaaccta 1681 ccagagcctagatgagacaccgaattaacattaaaatttcagtaactgactgtccctcat 1741 gtccatggcctaccatcccttctgaccctggcttccagggacctatgtcttttaatactc 1801 actgtcacattgggcaaagttgcttctaatccttatttcccatgtgcacaagtctttttg 1861 tattccagcttcctgataacactgcttactgtggaatattcatttgacatctgtctcttt 1921 tcatttcttttaactaccatgcccttgatatatcttttgcacctgctgaacttcatttct 1981 gtatcacctgacctctggatgccaaaacgtttattctgctttgtctgttgtagaatttta 2041 gataaagctattaatggcaatatttttttgctaaacgtttttgttttttactgtcactag 2101 ggcaataaaatttatactcaaccatataataacattttttaactactaaaggagtagttt 2161 ttattttaaagtcttagcaatttctattacaacttttcttagacttaacacttatgataa 2221 atgactaacatagtaacagaatctttatgaaatatgaccttttctgaaaatacatacttt 2281 tacatttctactttattgagacctattagatgtaagtgctagtagaatataagataaaag 2341 aggctgagaattaccatacaagggtattacaactgtaaaacaatttatctttgtttcatt 2401 gttctgtcaataattgttaccaaagagataaaaataaaagcagaatgtatatcatcccat 2461 ctgaaaaacactaattattgacatgtgcatctgtacaataaacttaaaatgattattaaa 2521 taatcaaatatatctactacattgtttatattattgaataaagtatattttccaaatgta 2581 aaaaaaaaaaaa HumanIL-33mRNAVariant3 (SEQIDNO:98) 1 agtctacagactcctccgaacacagagctgcagctcttcagggaagaaatcaaaacaaga 61 tcacaagaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccaca 121 gcaaagtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaataaggtgtta 181 ctgagttactatgagtctcaacacccctcaaatgaatcaggtgacggtgttgatggtaag 241 atgttaatggtaaccctgagtcctacaaaagacttctggttgcatgccaacaacaaggaa 301 cactctgtggagctccataagtgtgaaaaaccactgccagaccaggccttctttgtcctt 361 cataatatgcactccaactgtgtttcatttgaatgcaagactgatcctggagtgtttata 421 ggtgtaaaggataatcatcttgctctgattaaagtagactcttctgagaatttgtgtact 481 gaaaatatcttgtttaagctctctgaaacttagttgatggaaacctgtgagtcttgggtt 541 gagtacccaaatgctaccactggagaaggaatgagagataaagaaagagacaggtgacat 601 ctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatattatgtataaaaat 661 attttttctaatcctccagttattcttttatttccctctgtataactgcatcttcaatac 721 aagtatcagtatattaaatagggtattggtaaagaaacggtcaacattctaaagagatac 781 agtctgacctttacttttctctagtttcagtccagaaagaacttcatatttagagctaag 841 gccactgaggaaagagccatagcttaagtctctatgtagacagggatccattttaaagag 901 ctacttagagaaataattttccacagttccaaacgataggctcaaacactagagctgcta 961 gtaaaaagaagaccagatgcttcacagaattatcattttttcaactggaataaaacacca 1021 ggtttgtttgtagatgtcttaggcaacactcagagcagatctcccttactgtcaggggat 1081 atggaacttcaaaggcccacatggcaagccaggtaacataaatgtgtgaaaaagtaaaga 1141 taactaaaaaatttagaaaaataaatccagtatttgtaaagtgaataacttcatttctaa 1201 ttgtttaatttttaaaattctgatttttatatattgagtttaagcaaggcattcttacac 1261 gaggaagtgaagtaaattttagttcagacataaaatttcacttattaggaatatgtaaca 1321 tgctaaaacttttttttttttaaagagtactgagtcacaacatgttttagagcatccaag 1381 taccatataatccaactatcatggtaaggccagaaatcttctaacctaccagagcctaga 1441 tgagacaccgaattaacattaaaatttcagtaactgactgtccctcatgtccatggccta 1501 ccatcccttctgaccctggcttccagggacctatgtcttttaatactcactgtcacattg 1561 ggcaaagttgcttctaatccttatttcccatgtgcacaagtctttttgtattccagcttc 1621 ctgataacactgcttactgtggaatattcatttgacatctgtctcttttcatttctttta 1681 actaccatgcccttgatatatcttttgcacctgctgaacttcatttctgtatcacctgac 1741 ctctggatgccaaaacgtttattctgctttgtctgttgtagaattttagataaagctatt 1801 aatggcaatatttttttgctaaacgtttttgttttttactgtcactagggcaataaaatt 1861 tatactcaaccatataataacattttttaactactaaaggagtagtttttattttaaagt 1921 cttagcaatttctattacaacttttcttagacttaacacttatgataaatgactaacata 1981 gtaacagaatctttatgaaatatgaccttttctgaaaatacatacttttacatttctact 2041 ttattgagacctattagatgtaagtgctagtagaatataagataaaagaggctgagaatt 2101 accatacaagggtattacaactgtaaaacaatttatctttgtttcattgttctgtcaata 2161 attgttaccaaagagataaaaataaaagcagaatgtatatcatcccatctgaaaaacact 2221 aattattgacatgtgcatctgtacaataaacttaaaatgattattaaataatcaaatata 2281 tctactacattgtttatattattgaataaagtatattttccaaatgtaaaaaaaaaaaaa HumanIL-33mRNAVariant4 (SEQIDNO:99) 1 acagatgccaaacgagatggagagagggtgagtaggagcaaaatttctcatgagaatact 61 gaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccacagcaaagtggaaga 121 acacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacagaaggccaaagaag 181 tttgccccatgtactttatgaagctccgctctggccttatgataaaaaaggaggcctgtt 241 actttaggagagaaaccaccaaaaggccttcactgaaaacaggtagaaagcacaaaagac 301 atctggtactcgctgcctgtcaacagcagtctactgtggagtgctttgcctttggtatat 361 caggggtccagaaatatactagagcacttcatgattcaagtatcacaggaatttcaccta 421 ttacagagtatcttgcttctctaagcacatacaatgatcaatccattacttttgctttgg 481 aggatgaaagttatgagatatatgttgaagacttgaaaaaagatgaaaagaaagataagg 541 tgttactgagttactatgagtctcaacacccctcaaatgaatcaggtgacggtgttgatg 601 gtaagatgttaatggtaaccctgagtcctacaaaagacttctggttgcatgccaacaaca 661 aggaacactctgtggagctccataagtgtgaaaaaccactgccagaccaggccttctttg 721 tccttcataatatgcactccaactgtgtttcatttgaatgcaagactgatcctggagtgt 781 ttataggtgtaaaggataatcatcttgctctgattaaagtagactcttctgagaatttgt 841 gtactgaaaatatcttgtttaagctctctgaaacttagttgatggaaacctgtgagtctt 901 gggttgagtacccaaatgctaccactggagaaggaatgagagataaagaaagagacaggt 961 gacatctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatattatgtata 1021 aaaatattttttctaatcctccagttattcttttatttccctctgtataactgcatcttc 1081 aatacaagtatcagtatattaaatagggtattggtaaagaaacggtcaacattctaaaga 1141 gatacagtctgacctttacttttctctagtttcagtccagaaagaacttcatatttagag 1201 ctaaggccactgaggaaagagccatagcttaagtctctatgtagacagggatccatttta 1261 aagagctacttagagaaataattttccacagttccaaacgataggctcaaacactagagc 1321 tgctagtaaaaagaagaccagatgcttcacagaattatcattttttcaactggaataaaa 1381 caccaggtttgtttgtagatgtcttaggcaacactcagagcagatctcccttactgtcag 1441 gggatatggaacttcaaaggcccacatggcaagccaggtaacataaatgtgtgaaaaagt 1501 aaagataactaaaaaatttagaaaaataaatccagtatttgtaaagtgaataacttcatt 1561 tctaattgtttaatttttaaaattctgatttttatatattgagtttaagcaaggcattct 1621 tacacgaggaagtgaagtaaattttagttcagacataaaatttcacttattaggaatatg 1681 taacatgctaaaacttttttttttttaaagagtactgagtcacaacatgttttagagcat 1741 ccaagtaccatataatccaactatcatggtaaggccagaaatcttctaacctaccagagc 1801 ctagatgagacaccgaattaacattaaaatttcagtaactgactgtccctcatgtccatg 1861 gcctaccatcccttctgaccctggcttccagggacctatgtcttttaatactcactgtca 1921 cattgggcaaagttgcttctaatccttatttcccatgtgcacaagtctttttgtattcca 1981 gcttcctgataacactgcttactgtggaatattcatttgacatctgtctcttttcatttc 2041 ttttaactaccatgcccttgatatatcttttgcacctgctgaacttcatttctgtatcac 2101 ctgacctctggatgccaaaacgtttattctgctttgtctgttgtagaattttagataaag 2161 ctattaatggcaatatttttttgctaaacgtttttgttttttactgtcactagggcaata 2221 aaatttatactcaaccatataataacattttttaactactaaaggagtagtttttatttt 2281 aaagtcttagcaatttctattacaacttttcttagacttaacacttatgataaatgacta 2341 acatagtaacagaatctttatgaaatatgaccttttctgaaaatacatacttttacattt 2401 ctactttattgagacctattagatgtaagtgctagtagaatataagataaaagaggctga 2461 gaattaccatacaagggtattacaactgtaaaacaatttatctttgtttcattgttctgt 2521 caataattgttaccaaagagataaaaataaaagcagaatgtatatcatcccatctgaaaa 2581 acactaattattgacatgtgcatctgtacaataaacttaaaatgattattaaataatcaa 2641 atatatctactacattgtttatattattgaataaagtatattttccaaatgtaaaaaaaa 2701 aaaaa HumanIL-33mRNAVariant5 (SEQIDNO:100) 1 aaatactacaattgctgactacaggaaacctcatcatctgagaccagcactttataaatt 61 agaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccacagcaaa 121 gtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacagaaggc 181 caaagaagtttgccccatgtactttatgaagctccgctctggccttatgataaaaaagga 241 ggcctgttactttaggagagaaaccaccaaaaggccttcactgaaaacaggtagaaagca 301 caaaagacatctggtactcgctgcctgtcaacagcagtctactgtggagtgctttgcctt 361 tggtatatcaggggtccagaaatatactagagcacttcatgattcaagtatcacaggaat 421 ttcacctattacagagtatcttgcttctctaagcacatacaatgatcaatccattacttt 481 tgctttggaggatgaaagttatgagatatatgttgaagacttgaaaaaagatgaaaagaa 541 agataaggtgttactgagttactatgagtctcaacacccctcaaatgaatcaggtgacgg 601 tgttgatggtaagatgttaatggtaaccctgagtcctacaaaagacttctggttgcatgc 661 caacaacaaggaacactctgtggagctccataagtgtgaaaaaccactgccagaccaggc 721 cttctttgtccttcataatatgcactccaactgtgtttcatttgaatgcaagactgatcc 781 tggagtgtttataggtgtaaaggataatcatcttgctctgattaaagtagactcttctga 841 gaatttgtgtactgaaaatatcttgtttaagctctctgaaacttagttgatggaaacctg 901 tgagtcttgggttgagtacccaaatgctaccactggagaaggaatgagagataaagaaag 961 agacaggtgacatctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatat 1021 tatgtataaaaatattttttctaatcctccagttattcttttatttccctctgtataact 1081 gcatcttcaatacaagtatcagtatattaaatagggtattggtaaagaaacggtcaacat 1141 tctaaagagatacagtctgacctttacttttctctagtttcagtccagaaagaacttcat 1201 atttagagctaaggccactgaggaaagagccatagcttaagtctctatgtagacagggat 1261 ccattttaaagagctacttagagaaataattttccacagttccaaacgataggctcaaac 1321 actagagctgctagtaaaaagaagaccagatgcttcacagaattatcattttttcaactg 1381 gaataaaacaccaggtttgtttgtagatgtcttaggcaacactcagagcagatctccctt 1441 actgtcaggggatatggaacttcaaaggcccacatggcaagccaggtaacataaatgtgt 1501 gaaaaagtaaagataactaaaaaatttagaaaaataaatccagtatttgtaaagtgaata 1561 acttcatttctaattgtttaatttttaaaattctgatttttatatattgagtttaagcaa 1621 ggcattcttacacgaggaagtgaagtaaattttagttcagacataaaatttcacttatta 1681 ggaatatgtaacatgctaaaacttttttttttttaaagagtactgagtcacaacatgttt 1741 tagagcatccaagtaccatataatccaactatcatggtaaggccagaaatcttctaacct 1801 accagagcctagatgagacaccgaattaacattaaaatttcagtaactgactgtccctca 1861 tgtccatggcctaccatcccttctgaccctggcttccagggacctatgtcttttaatact 1921 cactgtcacattgggcaaagttgcttctaatccttatttcccatgtgcacaagtcttttt 1981 gtattccagcttcctgataacactgcttactgtggaatattcatttgacatctgtctctt 2041 ttcatttcttttaactaccatgcccttgatatatcttttgcacctgctgaacttcatttc 2101 tgtatcacctgacctctggatgccaaaacgtttattctgctttgtctgttgtagaatttt 2161 agataaagctattaatggcaatatttttttgctaaacgtttttgttttttactgtcacta 2221 gggcaataaaatttatactcaaccatataataacattttttaactactaaaggagtagtt 2281 tttattttaaagtcttagcaatttctattacaacttttcttagacttaacacttatgata 2341 aatgactaacatagtaacagaatctttatgaaatatgaccttttctgaaaatacatactt 2401 ttacatttctactttattgagacctattagatgtaagtgctagtagaatataagataaaa 2461 gaggctgagaattaccatacaagggtattacaactgtaaaacaatttatctttgtttcat 2521 tgttctgtcaataattgttaccaaagagataaaaataaaagcagaatgtatatcatccca 2581 tctgaaaaacactaattattgacatgtgcatctgtacaataaacttaaaatgattattaa 2641 ataatcaaatatatctactacattgtttatattattgaataaagtatattttccaaatgt 2701 aaaaaaaaaaaaa HumanIL-33mRNAVariant6 (SEQIDNO:101) 1 agtctacagactcctccgaacacagagctgcagctcttcagggaagaaatcaaaacaaga 61 tcacaagaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccaca 121 gcaaagtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacag 181 aaggccaaagaagtttgccccatgtactttatgaagctccgctctggccttatgataaaa 241 aaggaggcctgttactttaggagagaaaccaccaaaaggccttcactgaaaacaggtaga 301 aagcacaaaagacatctggtactcgctgcctgtcaacagcagtctactgtggagtgcttt 361 gcctttggtatatcaggggtccagaaatatactagagcacttcatgattcaagtatcaca 421 gagtatcttgcttctctaagcacatacaatgatcaatccattacttttgctttggaggat 481 gaaagttatgagatatatgttgaagacttgaaaaaagatgaaaagaaagataaggtgtta 541 ctgagttactatgagtctcaacacccctcaaatgaatcaggtgacggtgttgatggtaag 601 atgttaatggtaaccctgagtcctacaaaagacttctggttgcatgccaacaacaaggaa 661 cactctgtggagctccataagtgtgaaaaaccactgccagaccaggccttctttgtcctt 721 cataatatgcactccaactgtgtttcatttgaatgcaagactgatcctggagtgtttata 781 ggtgtaaaggataatcatcttgctctgattaaagtagactcttctgagaatttgtgtact 841 gaaaatatcttgtttaagctctctgaaacttagttgatggaaacctgtgagtcttgggtt 901 gagtacccaaatgctaccactggagaaggaatgagagataaagaaagagacaggtgacat 961 ctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatattatgtataaaaat 1021 attttttctaatcctccagttattcttttatttccctctgtataactgcatcttcaatac 1081 aagtatcagtatattaaatagggtattggtaaagaaacggtcaacattctaaagagatac 1141 agtctgacctttacttttctctagtttcagtccagaaagaacttcatatttagagctaag 1201 gccactgaggaaagagccatagcttaagtctctatgtagacagggatccattttaaagag 1261 ctacttagagaaataattttccacagttccaaacgataggctcaaacactagagctgcta 1321 gtaaaaagaagaccagatgcttcacagaattatcattttttcaactggaataaaacacca 1381 ggtttgtttgtagatgtcttaggcaacactcagagcagatctcccttactgtcaggggat 1441 atggaacttcaaaggcccacatggcaagccaggtaacataaatgtgtgaaaaagtaaaga 1501 taactaaaaaatttagaaaaataaatccagtatttgtaaagtgaataacttcatttctaa 1561 ttgtttaatttttaaaattctgatttttatatattgagtttaagcaaggcattcttacac 1621 gaggaagtgaagtaaattttagttcagacataaaatttcacttattaggaatatgtaaca 1681 tgctaaaacttttttttttttaaagagtactgagtcacaacatgttttagagcatccaag 1741 taccatataatccaactatcatggtaaggccagaaatcttctaacctaccagagcctaga 1801 tgagacaccgaattaacattaaaatttcagtaactgactgtccctcatgtccatggccta 1861 ccatcccttctgaccctggcttccagggacctatgtcttttaatactcactgtcacattg 1921 ggcaaagttgcttctaatccttatttcccatgtgcacaagtctttttgtattccagcttc 1981 ctgataacactgcttactgtggaatattcatttgacatctgtctcttttcatttctttta 2041 actaccatgcccttgatatatcttttgcacctgctgaacttcatttctgtatcacctgac 2101 ctctggatgccaaaacgtttattctgctttgtctgttgtagaattttagataaagctatt 2161 aatggcaatatttttttgctaaacgtttttgttttttactgtcactagggcaataaaatt 2221 tatactcaaccatataataacattttttaactactaaaggagtagtttttattttaaagt 2281 cttagcaatttctattacaacttttcttagacttaacacttatgataaatgactaacata 2341 gtaacagaatctttatgaaatatgaccttttctgaaaatacatacttttacatttctact 2401 ttattgagacctattagatgtaagtgctagtagaatataagataaaagaggctgagaatt 2461 accatacaagggtattacaactgtaaaacaatttatctttgtttcattgttctgtcaata 2521 attgttaccaaagagataaaaataaaagcagaatgtatatcatcccatctgaaaaacact 2581 aattattgacatgtgcatctgtacaataaacttaaaatgattattaaataatcaaatata 2641 tctactacattgtttatattattgaataaagtatattttccaaatgtaaaaaaaaaaaaa HumanIL-33mRNAVariant7 (SEQIDNO:102) 1 acagatgccaaacgagatggagagagggtgagtaggagcaaaatttctcatgagaatact 61 gaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccacagcaaagtggaaga 121 acacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacagaaggccaaagaag 181 tttgccccatgtactttatgaagctccgctctggccttatgataaaaaaggaggcctgtt 241 actttaggagagaaaccaccaaaaggccttcactgaaaacaggtagaaagcacaaaagac 301 atctggtactcgctgcctgtcaacagcagtctactgtggagtgctttgcctttggtatat 361 caggggtccagaaatatactagagcacttcatgattcaagtatcacagagtatcttgctt 421 ctctaagcacatacaatgatcaatccattacttttgctttggaggatgaaagttatgaga 481 tatatgttgaagacttgaaaaaagatgaaaagaaagataaggtgttactgagttactatg 541 agtctcaacacccctcaaatgaatcaggtgacggtgttgatggtaagatgttaatggtaa 601 ccctgagtcctacaaaagacttctggttgcatgccaacaacaaggaacactctgtggagc 661 tccataagtgtgaaaaaccactgccagaccaggccttctttgtccttcataatatgcact 721 ccaactgtgtttcatttgaatgcaagactgatcctggagtgtttataggtgtaaaggata 781 atcatcttgctctgattaaagtagactcttctgagaatttgtgtactgaaaatatcttgt 841 ttaagctctctgaaacttagttgatggaaacctgtgagtcttgggttgagtacccaaatg 901 ctaccactggagaaggaatgagagataaagaaagagacaggtgacatctaagggaaatga 961 agagtgcttagcatgtgtggaatgttttccatattatgtataaaaatattttttctaatc 1021 ctccagttattcttttatttccctctgtataactgcatcttcaatacaagtatcagtata 1081 ttaaatagggtattggtaaagaaacggtcaacattctaaagagatacagtctgaccttta 1141 cttttctctagtttcagtccagaaagaacttcatatttagagctaaggccactgaggaaa 1201 gagccatagcttaagtctctatgtagacagggatccattttaaagagctacttagagaaa 1261 taattttccacagttccaaacgataggctcaaacactagagctgctagtaaaaagaagac 1321 cagatgcttcacagaattatcattttttcaactggaataaaacaccaggtttgtttgtag 1381 atgtcttaggcaacactcagagcagatctcccttactgtcaggggatatggaacttcaaa 1441 ggcccacatggcaagccaggtaacataaatgtgtgaaaaagtaaagataactaaaaaatt 1501 tagaaaaataaatccagtatttgtaaagtgaataacttcatttctaattgtttaattttt 1561 aaaattctgatttttatatattgagtttaagcaaggcattcttacacgaggaagtgaagt 1621 aaattttagttcagacataaaatttcacttattaggaatatgtaacatgctaaaactttt 1681 ttttttttaaagagtactgagtcacaacatgttttagagcatccaagtaccatataatcc 1741 aactatcatggtaaggccagaaatcttctaacctaccagagcctagatgagacaccgaat 1801 taacattaaaatttcagtaactgactgtccctcatgtccatggcctaccatcccttctga 1861 ccctggcttccagggacctatgtcttttaatactcactgtcacattgggcaaagttgctt 1921 ctaatccttatttcccatgtgcacaagtctttttgtattccagcttcctgataacactgc 1981 ttactgtggaatattcatttgacatctgtctcttttcatttcttttaactaccatgccct 2041 tgatatatcttttgcacctgctgaacttcatttctgtatcacctgacctctggatgccaa 2101 aacgtttattctgctttgtctgttgtagaattttagataaagctattaatggcaatattt 2161 ttttgctaaacgtttttgttttttactgtcactagggcaataaaatttatactcaaccat 2221 ataataacattttttaactactaaaggagtagtttttattttaaagtcttagcaatttct 2281 attacaacttttcttagacttaacacttatgataaatgactaacatagtaacagaatctt 2341 tatgaaatatgaccttttctgaaaatacatacttttacatttctactttattgagaccta 2401 ttagatgtaagtgctagtagaatataagataaaagaggctgagaattaccatacaagggt 2461 attacaactgtaaaacaatttatctttgtttcattgttctgtcaataattgttaccaaag 2521 agataaaaataaaagcagaatgtatatcatcccatctgaaaaacactaattattgacatg 2581 tgcatctgtacaataaacttaaaatgattattaaataatcaaatatatctactacattgt 2641 ttatattattgaataaagtatattttccaaatgtaaaaaaaaaaaaa HumanIL-33mRNAVariant8 (SEQIDNO:103) 1 agtctacagactcctccgaacacagagctgcagctcttcagggaagaaatcaaaacaaga 61 tcacaagaatactgaaaaatgaagcctaaaatgaagtattcaaccaacaaaatttccaca 121 gcaaagtggaagaacacagcaagcaaagccttgtgtttcaagctgggaaaatcccaacag 181 aaggccaaagaagtttgccccatgtactttatgaagctccgctctggccttatgataaaa 241 aaggaggcctgttactttaggagagaaaccaccaaaaggccttcactgaaaacaggaatt 301 tcacctattacagagtatcttgcttctctaagcacatacaatgatcaatccattactttt 361 gctttggaggatgaaagttatgagatatatgttgaagacttgaaaaaagatgaaaagaaa 421 gataaggtgttactgagttactatgagtctcaacacccctcaaatgaatcaggtgacggt 481 gttgatggtaagatgttaatggtaaccctgagtcctacaaaagacttctggttgcatgcc 541 aacaacaaggaacactctgtggagctccataagtgtgaaaaaccactgccagaccaggcc 601 ttctttgtccttcataatatgcactccaactgtgtttcatttgaatgcaagactgatcct 661 ggagtgtttataggtgtaaaggataatcatcttgctctgattaaagtagactcttctgag 721 aatttgtgtactgaaaatatcttgtttaagctctctgaaacttagttgatggaaacctgt 781 gagtcttgggttgagtacccaaatgctaccactggagaaggaatgagagataaagaaaga 841 gacaggtgacatctaagggaaatgaagagtgcttagcatgtgtggaatgttttccatatt 901 atgtataaaaatattttttctaatcctccagttattcttttatttccctctgtataactg 961 catcttcaatacaagtatcagtatattaaatagggtattggtaaagaaacggtcaacatt 1021 ctaaagagatacagtctgacctttacttttctctagtttcagtccagaaagaacttcata 1081 tttagagctaaggccactgaggaaagagccatagcttaagtctctatgtagacagggatc 1141 cattttaaagagctacttagagaaataattttccacagttccaaacgataggctcaaaca 1201 ctagagctgctagtaaaaagaagaccagatgcttcacagaattatcattttttcaactgg 1261 aataaaacaccaggtttgtttgtagatgtcttaggcaacactcagagcagatctccctta 1321 ctgtcaggggatatggaacttcaaaggcccacatggcaagccaggtaacataaatgtgtg 1381 aaaaagtaaagataactaaaaaatttagaaaaataaatccagtatttgtaaagtgaataa 1441 cttcatttctaattgtttaatttttaaaattctgatttttatatattgagtttaagcaag 1501 gcattcttacacgaggaagtgaagtaaattttagttcagacataaaatttcacttattag 1561 gaatatgtaacatgctaaaacttttttttttttaaagagtactgagtcacaacatgtttt 1621 agagcatccaagtaccatataatccaactatcatggtaaggccagaaatcttctaaccta 1681 ccagagcctagatgagacaccgaattaacattaaaatttcagtaactgactgtccctcat 1741 gtccatggcctaccatcccttctgaccctggcttccagggacctatgtcttttaatactc 1801 actgtcacattgggcaaagttgcttctaatccttatttcccatgtgcacaagtctttttg 1861 tattccagcttcctgataacactgcttactgtggaatattcatttgacatctgtctcttt 1921 tcatttcttttaactaccatgcccttgatatatcttttgcacctgctgaacttcatttct 1981 gtatcacctgacctctggatgccaaaacgtttattctgctttgtctgttgtagaatttta 2041 gataaagctattaatggcaatatttttttgctaaacgtttttgttttttactgtcactag 2101 ggcaataaaatttatactcaaccatataataacattttttaactactaaaggagtagttt 2161 ttattttaaagtcttagcaatttctattacaacttttcttagacttaacacttatgataa 2221 atgactaacatagtaacagaatctttatgaaatatgaccttttctgaaaatacatacttt 2281 tacatttctactttattgagacctattagatgtaagtgctagtagaatataagataaaag 2341 aggctgagaattaccatacaagggtattacaactgtaaaacaatttatctttgtttcatt 2401 gttctgtcaataattgttaccaaagagataaaaataaaagcagaatgtatatcatcccat 2461 ctgaaaaacactaattattgacatgtgcatctgtacaataaacttaaaatgattattaaa 2521 taatcaaatatatctactacattgtttatattattgaataaagtatattttccaaatgta 2581 aaaaaaaaaaaa HumanIL-1R1mRNAVariant1 (SEQIDNO:104) 1 gtggccggcggccggagccgactcggagcgcgcggcgccggccgggaggagccggagagc 61 ggccgggccgggcggtgggggcgccggcctgccccgcgcgccccagggagcggcaggaat 121 gtgacaatcgcgcgcccgcgcaccgaagcactcctcgctcggctcctagggctctcgccc 181 ctctgagctgagccgggttccgcccggggctgggatcccatcaccctccacggccgtccg 241 tccaggtagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggt 301 aaaagacaaggccttctccaagaagaatatgaaagtgttactcagacttatttgtttcat 361 agctctactgatttcttctctggaggctgataaatgcaaggaacgtgaagaaaaaataat 421 tttagtgtcatctgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaaca 481 caaaggcactataacttggtataaagatgacagcaagacacctgtatctacagaacaagc 541 ctccaggattcatcaacacaaagagaaactttggtttgttcctgctaaggtggaggattc 601 aggacattactattgcgtggtaagaaattcatcttactgcctcagaattaaaataagtgc 661 aaaatttgtggagaatgagcctaacttatgttataatgcacaagccatatttaagcagaa 721 actacccgttgcaggagacggaggacttgtgtgcccttatatggagttttttaaaaatga 781 aaataatgagttacctaaattacagtggtataaggattgcaaacctctacttcttgacaa 841 tatacactttagtggagtcaaagataggctcatcgtgatgaatgtggctgaaaagcatag 901 agggaactatacttgtcatgcatcctacacatacttgggcaagcaatatcctattacccg 961 ggtaatagaatttattactctagaggaaaacaaacccacaaggcctgtgattgtgagccc 1021 agctaatgagacaatggaagtagacttgggatcccagatacaattgatctgtaatgtcac 1081 cggccagttgagtgacattgcttactggaagtggaatgggtcagtaattgatgaagatga 1141 cccagtgctaggggaagactattacagtgtggaaaatcctgcaaacaaaagaaggagtac 1201 cctcatcacagtgcttaatatatcggaaattgaaagtagattttataaacatccatttac 1261 ctgttttgccaagaatacacatggtatagatgcagcatatatccagttaatatatccagt 1321 cactaatttccagaagcacatgattggtatatgtgtcacgttgacagtcataattgtgtg 1381 ttctgttttcatctataaaatcttcaagattgacattgtgctttggtacagggattcctg 1441 ctatgattttctcccaataaaagcttcagatggaaagacctatgacgcatatatactgta 1501 tccaaagactgttggggaagggtctacctctgactgtgatatttttgtgtttaaagtctt 1561 gcctgaggtcttggaaaaacagtgtggatataagctgttcatttatggaagggatgacta 1621 cgttggggaagacattgttgaggtcattaatgaaaacgtaaagaaaagcagaagactgat 1681 tatcattttagtcagagaaacatcaggcttcagctggctgggtggttcatctgaagagca 1741 aatagccatgtataatgctcttgttcaggatggaattaaagttgtcctgcttgagctgga 1801 gaaaatccaagactatgagaaaatgccagaatcgattaaattcattaagcagaaacatgg 1861 ggctatccgctggtcaggggactttacacagggaccacagtctgcaaagacaaggttctg 1921 gaagaatgtcaggtaccacatgccagtccagcgacggtcaccttcatctaaacaccagtt 1981 actgtcaccagccactaaggagaaactgcaaagagaggctcacgtgcctctcgggtagca 2041 tggagaagttgccaagagttctttaggtgcctcctgtcttatggcgttgcaggccaggtt 2101 atgcctcatgctgacttgcagagttcatggaatgtaactatatcatcctttatccctgag 2161 gtcacctggaatcagattattaagggaataagccatgacgtcaatagcagcccagggcac 2221 ttcagagtagagggcttgggaagatcttttaaaaaggcagtaggcccggtgtggtggctc 2281 acgcctataatcccagcactttgggaggctgaagtgggtggatcaccagaggtcaggagt 2341 tcgagaccagcccagccaacatggcaaaaccccatctctactaaaaatacaaaaatgagc 2401 taggcatggtggcacacgcctgtaatcccagctacacctgaggctgaggcaggagaattg 2461 cttgaaccggggagacggaggttgcagtgagccgagtttgggccactgcactctagcctg 2521 gcaacagagcaagactccgtctcaaaaaaagggcaataaatgccctctctgaatgtttga 2581 actgccaagaaaaggcatggagacagcgaactagaagaaagggcaagaaggaaatagcca 2641 ccgtctacagatggcttagttaagtcatccacagcccaagggcggggctatgccttgtct 2701 ggggaccctgtagagtcactgaccctggagcggctctcctgagaggtgctgcaggcaaag 2761 tgagactgacacctcactgaggaagggagacatattcttggagaactttccatctgcttg 2821 tattttccatacacatccccagccagaagttagtgtccgaagaccgaattttattttaca 2881 gagcttgaaaactcacttcaatgaacaaagggattctccaggattccaaagttttgaagt 2941 catcttagctttccacaggagggagagaacttaaaaaagcaacagtagcagggaattgat 3001 ccacttcttaatgctttcctccctggcatgaccatcctgtcctttgttattatcctgcat 3061 tttacgtctttggaggaacagctccctagtggcttcctccgtctgcaatgtcccttgcac 3121 agcccacacatgaaccatccttcccatgatgccgctcttctgtcatcccgctcctgctga 3181 aacacctcccaggggctccacctgttcaggagctgaagcccatgctttcccaccagcatg 3241 tcactcccagaccacctccctgccctgtcctccagcttcccctcgctgtcctgctgtgtg 3301 aattcccaggttggcctggtggccatgtcgcctgcccccagcactcctctgtctctgctc 3361 ttgcctgcacccttcctcctcctttgcctaggaggccttctcgcattttctctagctgat 3421 cagaattttaccaaaattcagaacatcctccaattccacagtctctgggagactttccct 3481 aagaggcgacttcctctccagccttctctctctggtcaggcccactgcagagatggtggt 3541 gagcacatctgggaggctggtctccctccagctggaattgctgctctctgagggagaggc 3601 tgtggtggctgtctctgtccctcactgccttccaggagcaatttgcacatgtaacataga 3661 tttatgtaatgctttatgtttaaaaacattccccaattatcttatttaatttttgcaatt 3721 attctaattttatatatagagaaagtgacctattttttaaaaaaatcacactctaagttc 3781 tattgaacctaggacttgagcctccatttctggcttctagtctggtgttctgagtacttg 3841 atttcaggtcaataacggtcccccctcactccacactggcacgtttgtgagaagaaatga 3901 cattttgctaggaagtgaccgagtctaggaatgcttttattcaagacaccaaattccaaa 3961 cttctaaatgttggaattttcaaaaattgtgtttagattttatgaaaaactcttctactt 4021 tcatctattctttccctagaggcaaacatttcttaaaatgtttcattttcattaaaaatg 4081 aaagccaaatttatatgccaccgattgcaggacacaagcacagttttaagagttgtatga 4141 acatggagaggacttttggtttttatatttctcgtatttaatatgggtgaacaccaactt 4201 ttatttggaataataattttcctcctaaacaaaaacacattgagtttaagtctctgactc 4261 ttgcctttccacctgctttctcctgggcccgctttgcctgcttgaaggaacagtgctgtt 4321 ctggagctgctgttccaacagacagggcctagctttcatttgacacacagactacagcca 4381 gaagcccatggagcagggatgtcacgtcttgaaaagcctattagatgttttacaaattta 4441 attttgcagattattttagtctgtcatccagaaaatgtgtcagcatgcatagtgctaaga 4501 aagcaagccaatttggaaacttaggttagtgacaaaattggccagagagtgggggtgatg 4561 atgaccaagaattacaagtagaatggcagctggaatttaaggagggacaagaatcaatgg 4621 ataagcgtgggtggaggaagatccaaacagaaaagtgcaaagttattccccatcttccaa 4681 gggttgaattctggaggaagaagacacattcctagttccccgtgaacttcctttgactta 4741 ttgtccccactaaaacaaaacaaaaaacttttaatgccttccacattaattagattttct 4801 tgcagtttttttatggcatttttttaaagatgccctaagtgttgaagaagagtttgcaaa 4861 tgcaacaaaatatttaattaccggttgttaaaactggtttagcacaatttatattttccc 4921 tctcttgcctttcttatttgcaataaaaggtattgagccattttttaaatgacatttttg 4981 ataaattatgtttgtactagttgatgaaggagttttttttaacctgtttatataattttg 5041 cagcagaagccaaattttttgtatattaaagcaccaaattcatgtacagcatgcatcacg 5101 gatcaatagactgtacttattttccaataaaattttcaaactttgtactgttaaaaaaaa 5161 aaaaaaaaaa HumanIL-1R1mRNAVariant2 (SEQIDNO:105) 1 attggcagctcttcacttgtatcttttcatatcaaaaatgggaggtgacacccagtttaa 61 ggaaaattccaaggcatttgtctcgactaatgtgaaagatgattacagtggccagaggac 121 tgccaaggctccttctcaagctgcttgagtcaatgagggtagacgcaccctctgaagatg 181 gtgactccctcctgagaagctggaccccttggtaaaagacaaggccttctccaagaagaa 241 tatgaaagtgttactcagacttatttgtttcatagctctactgatttcttctctggaggc 301 tgataaatgcaaggaacgtgaagaaaaaataattttagtgtcatctgcaaatgaaattga 361 tgttcgtccctgtcctcttaacccaaatgaacacaaaggcactataacttggtataaaga 421 tgacagcaagacacctgtatctacagaacaagcctccaggattcatcaacacaaagagaa 481 actttggtttgttcctgctaaggtggaggattcaggacattactattgcgtggtaagaaa 541 ttcatcttactgcctcagaattaaaataagtgcaaaatttgtggagaatgagcctaactt 601 atgttataatgcacaagccatatttaagcagaaactacccgttgcaggagacggaggact 661 tgtgtgcccttatatggagttttttaaaaatgaaaataatgagttacctaaattacagtg 721 gtataaggattgcaaacctctacttcttgacaatatacactttagtggagtcaaagatag 781 gctcatcgtgatgaatgtggctgaaaagcatagagggaactatacttgtcatgcatccta 841 cacatacttgggcaagcaatatcctattacccgggtaatagaatttattactctagagga 901 aaacaaacccacaaggcctgtgattgtgagcccagctaatgagacaatggaagtagactt 961 gggatcccagatacaattgatctgtaatgtcaccggccagttgagtgacattgcttactg 1021 gaagtggaatgggtcagtaattgatgaagatgacccagtgctaggggaagactattacag 1081 tgtggaaaatcctgcaaacaaaagaaggagtaccctcatcacagtgcttaatatatcgga 1141 aattgaaagtagattttataaacatccatttacctgttttgccaagaatacacatggtat 1201 agatgcagcatatatccagttaatatatccagtcactaatttccagaagcacatgattgg 1261 tatatgtgtcacgttgacagtcataattgtgtgttctgttttcatctataaaatcttcaa 1321 gattgacattgtgctttggtacagggattcctgctatgattttctcccaataaaagtctt 1381 gcctgaggtcttggaaaaacagtgtggatataagctgttcatttatggaagggatgacta 1441 cgttggggaagacattgttgaggtcattaatgaaaacgtaaagaaaagcagaagactgat 1501 tatcattttagtcagagaaacatcaggcttcagctggctgggtggttcatctgaagagca 1561 aatagccatgtataatgctcttgttcaggatggaattaaagttgtcctgcttgagctgga 1621 gaaaatccaagactatgagaaaatgccagaatcgattaaattcattaagcagaaacatgg 1681 ggctatccgctggtcaggggactttacacagggaccacagtctgcaaagacaaggttctg 1741 gaagaatgtcaggtaccacatgccagtccagcgacggtcaccttcatctaaacaccagtt 1801 actgtcaccagccactaaggagaaactgcaaagagaggctcacgtgcctctcgggtagca 1861 tggagaagttgccaagagttctttaggtgcctcctgtcttatggcgttgcaggccaggtt 1921 atgcctcatgctgacttgcagagttcatggaatgtaactatatcatcctttatccctgag 1981 gtcacctggaatcagattattaagggaataagccatgacgtcaatagcagcccagggcac 2041 ttcagagtagagggcttgggaagatcttttaaaaaggcagtaggcccggtgtggtggctc 2101 acgcctataatcccagcactttgggaggctgaagtgggtggatcaccagaggtcaggagt 2161 tcgagaccagcccagccaacatggcaaaaccccatctctactaaaaatacaaaaatgagc 2221 taggcatggtggcacacgcctgtaatcccagctacacctgaggctgaggcaggagaattg 2281 cttgaaccggggagacggaggttgcagtgagccgagtttgggccactgcactctagcctg 2341 gcaacagagcaagactccgtctcaaaaaaagggcaataaatgccctctctgaatgtttga 2401 actgccaagaaaaggcatggagacagcgaactagaagaaagggcaagaaggaaatagcca 2461 ccgtctacagatggcttagttaagtcatccacagcccaagggcggggctatgccttgtct 2521 ggggaccctgtagagtcactgaccctggagcggctctcctgagaggtgctgcaggcaaag 2581 tgagactgacacctcactgaggaagggagacatattcttggagaactttccatctgcttg 2641 tattttccatacacatccccagccagaagttagtgtccgaagaccgaattttattttaca 2701 gagcttgaaaactcacttcaatgaacaaagggattctccaggattccaaagttttgaagt 2761 catcttagctttccacaggagggagagaacttaaaaaagcaacagtagcagggaattgat 2821 ccacttcttaatgctttcctccctggcatgaccatcctgtcctttgttattatcctgcat 2881 tttacgtctttggaggaacagctccctagtggcttcctccgtctgcaatgtcccttgcac 2941 agcccacacatgaaccatccttcccatgatgccgctcttctgtcatcccgctcctgctga 3001 aacacctcccaggggctccacctgttcaggagctgaagcccatgctttcccaccagcatg 3061 tcactcccagaccacctccctgccctgtcctccagcttcccctcgctgtcctgctgtgtg 3121 aattcccaggttggcctggtggccatgtcgcctgcccccagcactcctctgtctctgctc 3181 ttgcctgcacccttcctcctcctttgcctaggaggccttctcgcattttctctagctgat 3241 cagaattttaccaaaattcagaacatcctccaattccacagtctctgggagactttccct 3301 aagaggcgacttcctctccagccttctctctctggtcaggcccactgcagagatggtggt 3361 gagcacatctgggaggctggtctccctccagctggaattgctgctctctgagggagaggc 3421 tgtggtggctgtctctgtccctcactgccttccaggagcaatttgcacatgtaacataga 3481 tttatgtaatgctttatgtttaaaaacattccccaattatcttatttaatttttgcaatt 3541 attctaattttatatatagagaaagtgacctattttttaaaaaaatcacactctaagttc 3601 tattgaacctaggacttgagcctccatttctggcttctagtctggtgttctgagtacttg 3661 atttcaggtcaataacggtcccccctcactccacactggcacgtttgtgagaagaaatga 3721 cattttgctaggaagtgaccgagtctaggaatgcttttattcaagacaccaaattccaaa 3781 cttctaaatgttggaattttcaaaaattgtgtttagattttatgaaaaactcttctactt 3841 tcatctattctttccctagaggcaaacatttcttaaaatgtttcattttcattaaaaatg 3901 aaagccaaatttatatgccaccgattgcaggacacaagcacagttttaagagttgtatga 3961 acatggagaggacttttggtttttatatttctcgtatttaatatgggtgaacaccaactt 4021 ttatttggaataataattttcctcctaaacaaaaacacattgagtttaagtctctgactc 4081 ttgcctttccacctgctttctcctgggcccgctttgcctgcttgaaggaacagtgctgtt 4141 ctggagctgctgttccaacagacagggcctagctttcatttgacacacagactacagcca 4201 gaagcccatggagcagggatgtcacgtcttgaaaagcctattagatgttttacaaattta 4261 attttgcagattattttagtctgtcatccagaaaatgtgtcagcatgcatagtgctaaga 4321 aagcaagccaatttggaaacttaggttagtgacaaaattggccagagagtgggggtgatg 4381 atgaccaagaattacaagtagaatggcagctggaatttaaggagggacaagaatcaatgg 4441 ataagcgtgggtggaggaagatccaaacagaaaagtgcaaagttattccccatcttccaa 4501 gggttgaattctggaggaagaagacacattcctagttccccgtgaacttcctttgactta 4561 ttgtccccactaaaacaaaacaaaaaacttttaatgccttccacattaattagattttct 4621 tgcagtttttttatggcatttttttaaagatgccctaagtgttgaagaagagtttgcaaa 4681 tgcaacaaaatatttaattaccggttgttaaaactggtttagcacaatttatattttccc 4741 tctcttgcctttcttatttgcaataaaaggtattgagccattttttaaatgacatttttg 4801 ataaattatgtttgtactagttgatgaaggagttttttttaacctgtttatataattttg 4861 cagcagaagccaaattttttgtatattaaagcaccaaattcatgtacagcatgcatcacg 4921 gatcaatagactgtacttattttccaataaaattttcaaactttgtactgttaaaaaaaa 4981 aaaaaaaaaa HumanIL-1R1mRNAVariant3 (SEQIDNO:106) 1 attggcagctcttcacttgtatcttttcatatcaaaaatgggaggtgacacccagtttaa 61 ggaaaattccaaggcatttgtctcgactaatgtgaaagatgattacagtggccagaggac 121 tgccaaggctccttctcaagctgcttgagtcaatgagggtagacgcaccctctgaagatg 181 gtgactccctcctgagaagctggaccccttggtaaaagacaaggccttctccaagaagaa 241 tatgaaagtgttactcagacttatttgtttcatagctctactgatttcttctctggaggc 301 tgataaatgcaaggaacgtgaagaaaaaataattttagtgtcatctgcaaatgaaattga 361 tgttcgtccctgtcctcttaacccaaatgaacacaaaggcactataacttggtataaaga 421 tgacagcaagacacctgtatctacagaacaagcctccaggattcatcaacacaaagagaa 481 actttggtttgttcctgctaaggtggaggattcaggacattactattgcgtggtaagaaa 541 ttcatcttactgcctcagaattaaaataagtgcaaaatttgtggagaatgagcctaactt 601 atgttataatgcacaagccatatttaagcagaaactacccgttgcaggagacggaggact 661 tgtgtgcccttatatggagttttttaaaaatgaaaataatgagttacctaaattacagtg 721 gtataaggattgcaaacctctacttcttgacaatatacactttagtggagtcaaagatag 781 gctcatcgtgatgaatgtggctgaaaagcatagagggaactatacttgtcatgcatccta 841 cacatacttgggcaagcaatatcctattacccgggtaatagaatttattactctagagga 901 aaacaaacccacaaggcctgtgattgtgagcccagctaatgagacaatggaagtagactt 961 gggatcccagatacaattgatctgtaatgtcaccggccagttgagtgacattgcttactg 1021 gaagtggaatgggtcagtaattgatgaagatgacccagtgctaggggaagactattacag 1081 tgtggaaaatcctgcaaacaaaagaaggagtaccctcatcacagtgcttaatatatcgga 1141 aattgaaagtagattttataaacatccatttacctgttttgccaagaatacacatggtat 1201 agatgcagcatatatccagttaatatatccagtcactaatttccagaagcacatgattgg 1261 tatatgtgtcacgttgacagtcataattgtgtgttctgttttcatctataaaatcttcaa 1321 gattgacattgtgctttggtacagggattcctgctatgattttctcccaataaaagcttc 1381 agatggaaagacctatgacgcatatatactgtatccaaagactgttggggaagggtctac 1441 ctctgactgtgatatttttgtgtttaaagtcttgcctgaggtcttggaaaaacagtgtgg 1501 atataagctgttcatttatggaagggatgactacgttggggaagacattgttgaggtcat 1561 taatgaaaacgtaaagaaaagcagaagactgattatcattttagtcagagaaacatcagg 1621 cttcagctggctgggtggttcatctgaagagcaaatagccatgtataatgctcttgttca 1681 ggatggaattaaagttgtcctgcttgagctggagaaaatccaagactatgagaaaatgcc 1741 agaatcgattaaattcattaagcagaaacatggggctatccgctggtcaggggactttac 1801 acagggaccacagtctgcaaagacaaggttctggaagaatgtcaggtaccacatgccagt 1861 ccagcgacggtcaccttcatctaaacaccagttactgtcaccagccactaaggagaaact 1921 gcaaagagaggctcacgtgcctctcgggtagcatggagaagttgccaagagttctttagg 1981 tgcctcctgtcttatggcgttgcaggccaggttatgcctcatgctgacttgcagagttca 2041 tggaatgtaactatatcatcctttatccctgaggtcacctggaatcagattattaaggga 2101 ataagccatgacgtcaatagcagcccagggcacttcagagtagagggcttgggaagatct 2161 tttaaaaaggcagtaggcccggtgtggtggctcacgcctataatcccagcactttgggag 2221 gctgaagtgggtggatcaccagaggtcaggagttcgagaccagcccagccaacatggcaa 2281 aaccccatctctactaaaaatacaaaaatgagctaggcatggtggcacacgcctgtaatc 2341 ccagctacacctgaggctgaggcaggagaattgcttgaaccggggagacggaggttgcag 2401 tgagccgagtttgggccactgcactctagcctggcaacagagcaagactccgtctcaaaa 2461 aaagggcaataaatgccctctctgaatgtttgaactgccaagaaaaggcatggagacagc 2521 gaactagaagaaagggcaagaaggaaatagccaccgtctacagatggcttagttaagtca 2581 tccacagcccaagggcggggctatgccttgtctggggaccctgtagagtcactgaccctg 2641 gagcggctctcctgagaggtgctgcaggcaaagtgagactgacacctcactgaggaaggg 2701 agacatattcttggagaactttccatctgcttgtattttccatacacatccccagccaga 2761 agttagtgtccgaagaccgaattttattttacagagcttgaaaactcacttcaatgaaca 2821 aagggattctccaggattccaaagttttgaagtcatcttagctttccacaggagggagag 2881 aacttaaaaaagcaacagtagcagggaattgatccacttcttaatgctttcctccctggc 2941 atgaccatcctgtcctttgttattatcctgcattttacgtctttggaggaacagctccct 3001 agtggcttcctccgtctgcaatgtcccttgcacagcccacacatgaaccatccttcccat 3061 gatgccgctcttctgtcatcccgctcctgctgaaacacctcccaggggctccacctgttc 3121 aggagctgaagcccatgctttcccaccagcatgtcactcccagaccacctccctgccctg 3181 tcctccagcttcccctcgctgtcctgctgtgtgaattcccaggttggcctggtggccatg 3241 tcgcctgcccccagcactcctctgtctctgctcttgcctgcacccttcctcctcctttgc 3301 ctaggaggccttctcgcattttctctagctgatcagaattttaccaaaattcagaacatc 3361 ctccaattccacagtctctgggagactttccctaagaggcgacttcctctccagccttct 3421 ctctctggtcaggcccactgcagagatggtggtgagcacatctgggaggctggtctccct 3481 ccagctggaattgctgctctctgagggagaggctgtggtggctgtctctgtccctcactg 3541 ccttccaggagcaatttgcacatgtaacatagatttatgtaatgctttatgtttaaaaac 3601 attccccaattatcttatttaatttttgcaattattctaattttatatatagagaaagtg 3661 acctattttttaaaaaaatcacactctaagttctattgaacctaggacttgagcctccat 3721 ttctggcttctagtctggtgttctgagtacttgatttcaggtcaataacggtcccccctc 3781 actccacactggcacgtttgtgagaagaaatgacattttgctaggaagtgaccgagtcta 3841 ggaatgcttttattcaagacaccaaattccaaacttctaaatgttggaattttcaaaaat 3901 tgtgtttagattttatgaaaaactcttctactttcatctattctttccctagaggcaaac 3961 atttcttaaaatgtttcattttcattaaaaatgaaagccaaatttatatgccaccgattg 4021 caggacacaagcacagttttaagagttgtatgaacatggagaggacttttggtttttata 4081 tttctcgtatttaatatgggtgaacaccaacttttatttggaataataattttcctccta 4141 aacaaaaacacattgagtttaagtctctgactcttgcctttccacctgctttctcctggg 4201 cccgctttgcctgcttgaaggaacagtgctgttctggagctgctgttccaacagacaggg 4261 cctagctttcatttgacacacagactacagccagaagcccatggagcagggatgtcacgt 4321 cttgaaaagcctattagatgttttacaaatttaattttgcagattattttagtctgtcat 4381 ccagaaaatgtgtcagcatgcatagtgctaagaaagcaagccaatttggaaacttaggtt 4441 agtgacaaaattggccagagagtgggggtgatgatgaccaagaattacaagtagaatggc 4501 agctggaatttaaggagggacaagaatcaatggataagcgtgggtggaggaagatccaaa 4561 cagaaaagtgcaaagttattccccatcttccaagggttgaattctggaggaagaagacac 4621 attcctagttccccgtgaacttcctttgacttattgtccccactaaaacaaaacaaaaaa 4681 cttttaatgccttccacattaattagattttcttgcagtttttttatggcatttttttaa 4741 agatgccctaagtgttgaagaagagtttgcaaatgcaacaaaatatttaattaccggttg 4801 ttaaaactggtttagcacaatttatattttccctctcttgcctttcttatttgcaataaa 4861 aggtattgagccattttttaaatgacatttttgataaattatgtttgtactagttgatga 4921 aggagttttttttaacctgtttatataattttgcagcagaagccaaattttttgtatatt 4981 aaagcaccaaattcatgtacagcatgcatcacggatcaatagactgtacttattttccaa 5041 taaaattttcaaactttgtactgttaaaaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant4 (SEQIDNO:107) 1 attaaagccctaagaggctgtgacacagccatctccaaaaccccactttctccttccttt 61 gagcctccgtaccagctggggcgtccggcaagatgtgagttgtcactctgctgcggcaca 121 gacctgaattaacaactctagctagggctgacttcaaaaagcactttcgttttttaataa 181 ccaacatcagctcagcaggcttcatttgggaaaagaaaccttgtcggattaccccgacat 241 tctccacctcctgggaggccagccattcccaaatgccccaaggatgaagaacggagacgg 301 tagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggtaaaaga 361 caaggccttctccaagaagaatatgaaagtgttactcagacttatttgtttcatagctct 421 actgatttcttctctggaggctgataaatgcaaggaacgtgaagaaaaaataattttagt 481 gtcatctgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaacacaaagg 541 cactataacttggtataaagatgacagcaagacacctgtatctacagaacaagcctccag 601 gattcatcaacacaaagagaaactttggtttgttcctgctaaggtggaggattcaggaca 661 ttactattgcgtggtaagaaattcatcttactgcctcagaattaaaataagtgcaaaatt 721 tgtggagaatgagcctaacttatgttataatgcacaagccatatttaagcagaaactacc 781 cgttgcaggagacggaggacttgtgtgcccttatatggagttttttaaaaatgaaaataa 841 tgagttacctaaattacagtggtataaggattgcaaacctctacttcttgacaatataca 901 ctttagtggagtcaaagataggctcatcgtgatgaatgtggctgaaaagcatagagggaa 961 ctatacttgtcatgcatcctacacatacttgggcaagcaatatcctattacccgggtaat 1021 agaatttattactctagaggaaaacaaacccacaaggcctgtgattgtgagcccagctaa 1081 tgagacaatggaagtagacttgggatcccagatacaattgatctgtaatgtcaccggcca 1141 gttgagtgacattgcttactggaagtggaatgggtcagtaattgatgaagatgacccagt 1201 gctaggggaagactattacagtgtggaaaatcctgcaaacaaaagaaggagtaccctcat 1261 cacagtgcttaatatatcggaaattgaaagtagattttataaacatccatttacctgttt 1321 tgccaagaatacacatggtatagatgcagcatatatccagttaatatatccagtcactaa 1381 tttccagaagcacatgattggtatatgtgtcacgttgacagtcataattgtgtgttctgt 1441 tttcatctataaaatcttcaagattgacattgtgctttggtacagggattcctgctatga 1501 ttttctcccaataaaagcttcagatggaaagacctatgacgcatatatactgtatccaaa 1561 gactgttggggaagggtctacctctgactgtgatatttttgtgtttaaagtcttgcctga 1621 ggtcttggaaaaacagtgtggatataagctgttcatttatggaagggatgactacgttgg 1681 ggaagacattgttgaggtcattaatgaaaacgtaaagaaaagcagaagactgattatcat 1741 tttagtcagagaaacatcaggcttcagctggctgggtggttcatctgaagagcaaatagc 1801 catgtataatgctcttgttcaggatggaattaaagttgtcctgcttgagctggagaaaat 1861 ccaagactatgagaaaatgccagaatcgattaaattcattaagcagaaacatggggctat 1921 ccgctggtcaggggactttacacagggaccacagtctgcaaagacaaggttctggaagaa 1981 tgtcaggtaccacatgccagtccagcgacggtcaccttcatctaaacaccagttactgtc 2041 accagccactaaggagaaactgcaaagagaggctcacgtgcctctcgggtagcatggaga 2101 agttgccaagagttctttaggtgcctcctgtcttatggcgttgcaggccaggttatgcct 2161 catgctgacttgcagagttcatggaatgtaactatatcatcctttatccctgaggtcacc 2221 tggaatcagattattaagggaataagccatgacgtcaatagcagcccagggcacttcaga 2281 gtagagggcttgggaagatcttttaaaaaggcagtaggcccggtgtggtggctcacgcct 2341 ataatcccagcactttgggaggctgaagtgggtggatcaccagaggtcaggagttcgaga 2401 ccagcccagccaacatggcaaaaccccatctctactaaaaatacaaaaatgagctaggca 2461 tggtggcacacgcctgtaatcccagctacacctgaggctgaggcaggagaattgcttgaa 2521 ccggggagacggaggttgcagtgagccgagtttgggccactgcactctagcctggcaaca 2581 gagcaagactccgtctcaaaaaaagggcaataaatgccctctctgaatgtttgaactgcc 2641 aagaaaaggcatggagacagcgaactagaagaaagggcaagaaggaaatagccaccgtct 2701 acagatggcttagttaagtcatccacagcccaagggcggggctatgccttgtctggggac 2761 cctgtagagtcactgaccctggagcggctctcctgagaggtgctgcaggcaaagtgagac 2821 tgacacctcactgaggaagggagacatattcttggagaactttccatctgcttgtatttt 2881 ccatacacatccccagccagaagttagtgtccgaagaccgaattttattttacagagctt 2941 gaaaactcacttcaatgaacaaagggattctccaggattccaaagttttgaagtcatctt 3001 agctttccacaggagggagagaacttaaaaaagcaacagtagcagggaattgatccactt 3061 cttaatgctttcctccctggcatgaccatcctgtcctttgttattatcctgcattttacg 3121 tctttggaggaacagctccctagtggcttcctccgtctgcaatgtcccttgcacagccca 3181 cacatgaaccatccttcccatgatgccgctcttctgtcatcccgctcctgctgaaacacc 3241 tcccaggggctccacctgttcaggagctgaagcccatgctttcccaccagcatgtcactc 3301 ccagaccacctccctgccctgtcctccagcttcccctcgctgtcctgctgtgtgaattcc 3361 caggttggcctggtggccatgtcgcctgcccccagcactcctctgtctctgctcttgcct 3421 gcacccttcctcctcctttgcctaggaggccttctcgcattttctctagctgatcagaat 3481 tttaccaaaattcagaacatcctccaattccacagtctctgggagactttccctaagagg 3541 cgacttcctctccagccttctctctctggtcaggcccactgcagagatggtggtgagcac 3601 atctgggaggctggtctccctccagctggaattgctgctctctgagggagaggctgtggt 3661 ggctgtctctgtccctcactgccttccaggagcaatttgcacatgtaacatagatttatg 3721 taatgctttatgtttaaaaacattccccaattatcttatttaatttttgcaattattcta 3781 attttatatatagagaaagtgacctattttttaaaaaaatcacactctaagttctattga 3841 acctaggacttgagcctccatttctggcttctagtctggtgttctgagtacttgatttca 3901 ggtcaataacggtcccccctcactccacactggcacgtttgtgagaagaaatgacatttt 3961 gctaggaagtgaccgagtctaggaatgcttttattcaagacaccaaattccaaacttcta 4021 aatgttggaattttcaaaaattgtgtttagattttatgaaaaactcttctactttcatct 4081 attctttccctagaggcaaacatttcttaaaatgtttcattttcattaaaaatgaaagcc 4141 aaatttatatgccaccgattgcaggacacaagcacagttttaagagttgtatgaacatgg 4201 agaggacttttggtttttatatttctcgtatttaatatgggtgaacaccaacttttattt 4261 ggaataataattttcctcctaaacaaaaacacattgagtttaagtctctgactcttgcct 4321 ttccacctgctttctcctgggcccgctttgcctgcttgaaggaacagtgctgttctggag 4381 ctgctgttccaacagacagggcctagctttcatttgacacacagactacagccagaagcc 4441 catggagcagggatgtcacgtcttgaaaagcctattagatgttttacaaatttaattttg 4501 cagattattttagtctgtcatccagaaaatgtgtcagcatgcatagtgctaagaaagcaa 4561 gccaatttggaaacttaggttagtgacaaaattggccagagagtgggggtgatgatgacc 4621 aagaattacaagtagaatggcagctggaatttaaggagggacaagaatcaatggataagc 4681 gtgggtggaggaagatccaaacagaaaagtgcaaagttattccccatcttccaagggttg 4741 aattctggaggaagaagacacattcctagttccccgtgaacttcctttgacttattgtcc 4801 ccactaaaacaaaacaaaaaacttttaatgccttccacattaattagattttcttgcagt 4861 ttttttatggcatttttttaaagatgccctaagtgttgaagaagagtttgcaaatgcaac 4921 aaaatatttaattaccggttgttaaaactggtttagcacaatttatattttccctctctt 4981 gcctttcttatttgcaataaaaggtattgagccattttttaaatgacatttttgataaat 5041 tatgtttgtactagttgatgaaggagttttttttaacctgtttatataattttgcagcag 5101 aagccaaattttttgtatattaaagcaccaaattcatgtacagcatgcatcacggatcaa 5161 tagactgtacttattttccaataaaattttcaaactttgtactgttaaaaaaaaaaaaaa 5221 aaaa HumanIL-1R1mRNAVariant5 (SEQIDNO:108) 1 aggatggcccatgaagacctccaaacaagctggaggggccagtcacttgctgaagactag 61 cgaagtggagggggaaagcccgagggagctgcagactcgaccactgcgccctcccctcct 121 ctccctgcaaggagcccaaggtagacgcaccctctgaagatggtgactccctcctgagaa 181 gctggaccccttggtaaaagacaaggccttctccaagaagaatatgaaagtgttactcag 241 acttatttgtttcatagctctactgatttcttctctggaggctgataaatgcaaggaacg 301 tgaagaaaaaataattttagtgtcatctgcaaatgaaattgatgttcgtccctgtcctct 361 taacccaaatgaacacaaaggcactataacttggtataaagatgacagcaagacacctgt 421 atctacagaacaagcctccaggattcatcaacacaaagagaaactttggtttgttcctgc 481 taaggtggaggattcaggacattactattgcgtggtaagaaattcatcttactgcctcag 541 aattaaaataagtgcaaaatttgtggagaatgagcctaacttatgttataatgcacaagc 601 catatttaagcagaaactacccgttgcaggagacggaggacttgtgtgcccttatatgga 661 gttttttaaaaatgaaaataatgagttacctaaattacagtggtataaggattgcaaacc 721 tctacttcttgacaatatacactttagtggagtcaaagataggctcatcgtgatgaatgt 781 ggctgaaaagcatagagggaactatacttgtcatgcatcctacacatacttgggcaagca 841 atatcctattacccgggtaatagaatttattactctagaggaaaacaaacccacaaggcc 901 tgtgattgtgagcccagctaatgagacaatggaagtagacttgggatcccagatacaatt 961 gatctgtaatgtcaccggccagttgagtgacattgcttactggaagtggaatgggtcagt 1021 aattgatgaagatgacccagtgctaggggaagactattacagtgtggaaaatcctgcaaa 1081 caaaagaaggagtaccctcatcacagtgcttaatatatcggaaattgaaagtagatttta 1141 taaacatccatttacctgttttgccaagaatacacatggtatagatgcagcatatatcca 1201 gttaatatatccagtcactaatttccagaagcacatgattggtatatgtgtcacgttgac 1261 agtcataattgtgtgttctgttttcatctataaaatcttcaagattgacattgtgctttg 1321 gtacagggattcctgctatgattttctcccaataaaagcttcagatggaaagacctatga 1381 cgcatatatactgtatccaaagactgttggggaagggtctacctctgactgtgatatttt 1441 tgtgtttaaagtcttgcctgaggtcttggaaaaacagtgtggatataagctgttcattta 1501 tggaagggatgactacgttggggaagacattgttgaggtcattaatgaaaacgtaaagaa 1561 aagcagaagactgattatcattttagtcagagaaacatcaggcttcagctggctgggtgg 1621 ttcatctgaagagcaaatagccatgtataatgctcttgttcaggatggaattaaagttgt 1681 cctgcttgagctggagaaaatccaagactatgagaaaatgccagaatcgattaaattcat 1741 taagcagaaacatggggctatccgctggtcaggggactttacacagggaccacagtctgc 1801 aaagacaaggttctggaagaatgtcaggtaccacatgccagtccagcgacggtcaccttc 1861 atctaaacaccagttactgtcaccagccactaaggagaaactgcaaagagaggctcacgt 1921 gcctctcgggtagcatggagaagttgccaagagttctttaggtgcctcctgtcttatggc 1981 gttgcaggccaggttatgcctcatgctgacttgcagagttcatggaatgtaactatatca 2041 tcctttatccctgaggtcacctggaatcagattattaagggaataagccatgacgtcaat 2101 agcagcccagggcacttcagagtagagggcttgggaagatcttttaaaaaggcagtaggc 2161 ccggtgtggtggctcacgcctataatcccagcactttgggaggctgaagtgggtggatca 2221 ccagaggtcaggagttcgagaccagcccagccaacatggcaaaaccccatctctactaaa 2281 aatacaaaaatgagctaggcatggtggcacacgcctgtaatcccagctacacctgaggct 2341 gaggcaggagaattgcttgaaccggggagacggaggttgcagtgagccgagtttgggcca 2401 ctgcactctagcctggcaacagagcaagactccgtctcaaaaaaagggcaataaatgccc 2461 tctctgaatgtttgaactgccaagaaaaggcatggagacagcgaactagaagaaagggca 2521 agaaggaaatagccaccgtctacagatggcttagttaagtcatccacagcccaagggcgg 2581 ggctatgccttgtctggggaccctgtagagtcactgaccctggagcggctctcctgagag 2641 gtgctgcaggcaaagtgagactgacacctcactgaggaagggagacatattcttggagaa 2701 ctttccatctgcttgtattttccatacacatccccagccagaagttagtgtccgaagacc 2761 gaattttattttacagagcttgaaaactcacttcaatgaacaaagggattctccaggatt 2821 ccaaagttttgaagtcatcttagctttccacaggagggagagaacttaaaaaagcaacag 2881 tagcagggaattgatccacttcttaatgctttcctccctggcatgaccatcctgtccttt 2941 gttattatcctgcattttacgtctttggaggaacagctccctagtggcttcctccgtctg 3001 caatgtcccttgcacagcccacacatgaaccatccttcccatgatgccgctcttctgtca 3061 tcccgctcctgctgaaacacctcccaggggctccacctgttcaggagctgaagcccatgc 3121 tttcccaccagcatgtcactcccagaccacctccctgccctgtcctccagcttcccctcg 3181 ctgtcctgctgtgtgaattcccaggttggcctggtggccatgtcgcctgcccccagcact 3241 cctctgtctctgctcttgcctgcacccttcctcctcctttgcctaggaggccttctcgca 3301 ttttctctagctgatcagaattttaccaaaattcagaacatcctccaattccacagtctc 3361 tgggagactttccctaagaggcgacttcctctccagccttctctctctggtcaggcccac 3421 tgcagagatggtggtgagcacatctgggaggctggtctccctccagctggaattgctgct 3481 ctctgagggagaggctgtggtggctgtctctgtccctcactgccttccaggagcaatttg 3541 cacatgtaacatagatttatgtaatgctttatgtttaaaaacattccccaattatcttat 3601 ttaatttttgcaattattctaattttatatatagagaaagtgacctattttttaaaaaaa 3661 tcacactctaagttctattgaacctaggacttgagcctccatttctggcttctagtctgg 3721 tgttctgagtacttgatttcaggtcaataacggtcccccctcactccacactggcacgtt 3781 tgtgagaagaaatgacattttgctaggaagtgaccgagtctaggaatgcttttattcaag 3841 acaccaaattccaaacttctaaatgttggaattttcaaaaattgtgtttagattttatga 3901 aaaactcttctactttcatctattctttccctagaggcaaacatttcttaaaatgtttca 3961 ttttcattaaaaatgaaagccaaatttatatgccaccgattgcaggacacaagcacagtt 4021 ttaagagttgtatgaacatggagaggacttttggtttttatatttctcgtatttaatatg 4081 ggtgaacaccaacttttatttggaataataattttcctcctaaacaaaaacacattgagt 4141 ttaagtctctgactcttgcctttccacctgctttctcctgggcccgctttgcctgcttga 4201 aggaacagtgctgttctggagctgctgttccaacagacagggcctagctttcatttgaca 4261 cacagactacagccagaagcccatggagcagggatgtcacgtcttgaaaagcctattaga 4321 tgttttacaaatttaattttgcagattattttagtctgtcatccagaaaatgtgtcagca 4381 tgcatagtgctaagaaagcaagccaatttggaaacttaggttagtgacaaaattggccag 4441 agagtgggggtgatgatgaccaagaattacaagtagaatggcagctggaatttaaggagg 4501 gacaagaatcaatggataagcgtgggtggaggaagatccaaacagaaaagtgcaaagtta 4561 ttccccatcttccaagggttgaattctggaggaagaagacacattcctagttccccgtga 4621 acttcctttgacttattgtccccactaaaacaaaacaaaaaacttttaatgccttccaca 4681 ttaattagattttcttgcagtttttttatggcatttttttaaagatgccctaagtgttga 4741 agaagagtttgcaaatgcaacaaaatatttaattaccggttgttaaaactggtttagcac 4801 aatttatattttccctctcttgcctttcttatttgcaataaaaggtattgagccattttt 4861 taaatgacatttttgataaattatgtttgtactagttgatgaaggagttttttttaacct 4921 gtttatataattttgcagcagaagccaaattttttgtatattaaagcaccaaattcatgt 4981 acagcatgcatcacggatcaatagactgtacttattttccaataaaattttcaaactttg 5041 tactgttaaaaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant6 (SEQIDNO:109) 1 ctgatgccctggagtcgccaactcaattcgcgggtcgcagccaggctccatgggggtagt 61 agagccaggtcgtagtggctaggtagacgcaccctctgaagatggtgactccctcctgag 121 aagctggaccccttggtaaaagacaaggccttctccaagaagaatatgaaagtgttactc 181 agacttatttgtttcatagctctactgatttcttctctggaggctgataaatgcaaggaa 241 cgtgaagaaaaaataattttagtgtcatctgcaaatgaaattgatgttcgtccctgtcct 301 cttaacccaaatgaacacaaaggcactataacttggtataaagatgacagcaagacacct 361 gtatctacagaacaagcctccaggattcatcaacacaaagagaaactttggtttgttcct 421 gctaaggtggaggattcaggacattactattgcgtggtaagaaattcatcttactgcctc 481 agaattaaaataagtgcaaaatttgtggagaatgagcctaacttatgttataatgcacaa 541 gccatatttaagcagaaactacccgttgcaggagacggaggacttgtgtgcccttatatg 601 gagttttttaaaaatgaaaataatgagttacctaaattacagtggtataaggattgcaaa 661 cctctacttcttgacaatatacactttagtggagtcaaagataggctcatcgtgatgaat 721 gtggctgaaaagcatagagggaactatacttgtcatgcatcctacacatacttgggcaag 781 caatatcctattacccgggtaatagaatttattactctagaggaaaacaaacccacaagg 841 cctgtgattgtgagcccagctaatgagacaatggaagtagacttgggatcccagatacaa 901 ttgatctgtaatgtcaccggccagttgagtgacattgcttactggaagtggaatgggtca 961 gtaattgatgaagatgacccagtgctaggggaagactattacagtgtggaaaatcctgca 1021 aacaaaagaaggagtaccctcatcacagtgcttaatatatcggaaattgaaagtagattt 1081 tataaacatccatttacctgttttgccaagaatacacatggtatagatgcagcatatatc 1141 cagttaatatatccagtcactaatttccagaagcacatgattggtatatgtgtcacgttg 1201 acagtcataattgtgtgttctgttttcatctataaaatcttcaagattgacattgtgctt 1261 tggtacagggattcctgctatgattttctcccaataaaagcttcagatggaaagacctat 1321 gacgcatatatactgtatccaaagactgttggggaagggtctacctctgactgtgatatt 1381 tttgtgtttaaagtcttgcctgaggtcttggaaaaacagtgtggatataagctgttcatt 1441 tatggaagggatgactacgttggggaagacattgttgaggtcattaatgaaaacgtaaag 1501 aaaagcagaagactgattatcattttagtcagagaaacatcaggcttcagctggctgggt 1561 ggttcatctgaagagcaaatagccatgtataatgctcttgttcaggatggaattaaagtt 1621 gtcctgcttgagctggagaaaatccaagactatgagaaaatgccagaatcgattaaattc 1681 attaagcagaaacatggggctatccgctggtcaggggactttacacagggaccacagtct 1741 gcaaagacaaggttctggaagaatgtcaggtaccacatgccagtccagcgacggtcacct 1801 tcatctaaacaccagttactgtcaccagccactaaggagaaactgcaaagagaggctcac 1861 gtgcctctcgggtagcatggagaagttgccaagagttctttaggtgcctcctgtcttatg 1921 gcgttgcaggccaggttatgcctcatgctgacttgcagagttcatggaatgtaactatat 1981 catcctttatccctgaggtcacctggaatcagattattaagggaataagccatgacgtca 2041 atagcagcccagggcacttcagagtagagggcttgggaagatcttttaaaaaggcagtag 2101 gcccggtgtggtggctcacgcctataatcccagcactttgggaggctgaagtgggtggat 2161 caccagaggtcaggagttcgagaccagcccagccaacatggcaaaaccccatctctacta 2221 aaaatacaaaaatgagctaggcatggtggcacacgcctgtaatcccagctacacctgagg 2281 ctgaggcaggagaattgcttgaaccggggagacggaggttgcagtgagccgagtttgggc 2341 cactgcactctagcctggcaacagagcaagactccgtctcaaaaaaagggcaataaatgc 2401 cctctctgaatgtttgaactgccaagaaaaggcatggagacagcgaactagaagaaaggg 2461 caagaaggaaatagccaccgtctacagatggcttagttaagtcatccacagcccaagggc 2521 ggggctatgccttgtctggggaccctgtagagtcactgaccctggagcggctctcctgag 2581 aggtgctgcaggcaaagtgagactgacacctcactgaggaagggagacatattcttggag 2641 aactttccatctgcttgtattttccatacacatccccagccagaagttagtgtccgaaga 2701 ccgaattttattttacagagcttgaaaactcacttcaatgaacaaagggattctccagga 2761 ttccaaagttttgaagtcatcttagctttccacaggagggagagaacttaaaaaagcaac 2821 agtagcagggaattgatccacttcttaatgctttcctccctggcatgaccatcctgtcct 2881 ttgttattatcctgcattttacgtctttggaggaacagctccctagtggcttcctccgtc 2941 tgcaatgtcccttgcacagcccacacatgaaccatccttcccatgatgccgctcttctgt 3001 catcccgctcctgctgaaacacctcccaggggctccacctgttcaggagctgaagcccat 3061 gctttcccaccagcatgtcactcccagaccacctccctgccctgtcctccagcttcccct 3121 cgctgtcctgctgtgtgaattcccaggttggcctggtggccatgtcgcctgcccccagca 3181 ctcctctgtctctgctcttgcctgcacccttcctcctcctttgcctaggaggccttctcg 3241 cattttctctagctgatcagaattttaccaaaattcagaacatcctccaattccacagtc 3301 tctgggagactttccctaagaggcgacttcctctccagccttctctctctggtcaggccc 3361 actgcagagatggtggtgagcacatctgggaggctggtctccctccagctggaattgctg 3421 ctctctgagggagaggctgtggtggctgtctctgtccctcactgccttccaggagcaatt 3481 tgcacatgtaacatagatttatgtaatgctttatgtttaaaaacattccccaattatctt 3541 atttaatttttgcaattattctaattttatatatagagaaagtgacctattttttaaaaa 3601 aatcacactctaagttctattgaacctaggacttgagcctccatttctggcttctagtct 3661 ggtgttctgagtacttgatttcaggtcaataacggtcccccctcactccacactggcacg 3721 tttgtgagaagaaatgacattttgctaggaagtgaccgagtctaggaatgcttttattca 3781 agacaccaaattccaaacttctaaatgttggaattttcaaaaattgtgtttagattttat 3841 gaaaaactcttctactttcatctattctttccctagaggcaaacatttcttaaaatgttt 3901 cattttcattaaaaatgaaagccaaatttatatgccaccgattgcaggacacaagcacag 3961 ttttaagagttgtatgaacatggagaggacttttggtttttatatttctcgtatttaata 4021 tgggtgaacaccaacttttatttggaataataattttcctcctaaacaaaaacacattga 4081 gtttaagtctctgactcttgcctttccacctgctttctcctgggcccgctttgcctgctt 4141 gaaggaacagtgctgttctggagctgctgttccaacagacagggcctagctttcatttga 4201 cacacagactacagccagaagcccatggagcagggatgtcacgtcttgaaaagcctatta 4261 gatgttttacaaatttaattttgcagattattttagtctgtcatccagaaaatgtgtcag 4321 catgcatagtgctaagaaagcaagccaatttggaaacttaggttagtgacaaaattggcc 4381 agagagtgggggtgatgatgaccaagaattacaagtagaatggcagctggaatttaagga 4441 gggacaagaatcaatggataagcgtgggtggaggaagatccaaacagaaaagtgcaaagt 4501 tattccccatcttccaagggttgaattctggaggaagaagacacattcctagttccccgt 4561 gaacttcctttgacttattgtccccactaaaacaaaacaaaaaacttttaatgccttcca 4621 cattaattagattttcttgcagtttttttatggcatttttttaaagatgccctaagtgtt 4681 gaagaagagtttgcaaatgcaacaaaatatttaattaccggttgttaaaactggtttagc 4741 acaatttatattttccctctcttgcctttcttatttgcaataaaaggtattgagccattt 4801 tttaaatgacatttttgataaattatgtttgtactagttgatgaaggagttttttttaac 4861 ctgtttatataattttgcagcagaagccaaattttttgtatattaaagcaccaaattcat 4921 gtacagcatgcatcacggatcaatagactgtacttattttccaataaaattttcaaactt 4981 tgtactgttaaaaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant7 (SEQIDNO:110) 1 gtagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggtaaaag 61 acaaggccttctccaagataaatgcaaggaacgtgaagaaaaaataattttagtgtcatc 121 tgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaacacaaaggcactat 181 aacttggtataaagatgacagcaagacacctgtatctacagaacaagcctccaggattca 241 tcaacacaaagagaaactttggtttgttcctgctaaggtggaggattcaggacattacta 301 ttgcgtggtaagaaattcatcttactgcctcagaattaaaataagtgcaaaatttgtgga 361 gaatgagcctaacttatgttataatgcacaagccatatttaagcagaaactacccgttgc 421 aggagacggaggacttgtgtgcccttatatggagttttttaaaaatgaaaataatgagtt 481 acctaaattacagtggtataaggattgcaaacctctacttcttgacaatatacactttag 541 tggagtcaaagataggctcatcgtgatgaatgtggctgaaaagcatagagggaactatac 601 ttgtcatgcatcctacacatacttgggcaagcaatatcctattacccgggtaatagaatt 661 tattactctagaggaaaacaaacccacaaggcctgtgattgtgagcccagctaatgagac 721 aatggaagtagacttgggatcccagatacaattgatctgtaatgtcaccggccagttgag 781 tgacattgcttactggaagtggaatgggtcagtaattgatgaagatgacccagtgctagg 841 ggaagactattacagtgtggaaaatcctgcaaacaaaagaaggagtaccctcatcacagt 901 gcttaatatatcggaaattgaaagtagattttataaacatccatttacctgttttgccaa 961 gaatacacatggtatagatgcagcatatatccagttaatatatccagtcactaatttcca 1021 gaagcacatgattggtatatgtgtcacgttgacagtcataattgtgtgttctgttttcat 1081 ctataaaatcttcaagattgacattgtgctttggtacagggattcctgctatgattttct 1141 cccaataaaagcttcagatggaaagacctatgacgcatatatactgtatccaaagactgt 1201 tggggaagggtctacctctgactgtgatatttttgtgtttaaagtcttgcctgaggtctt 1261 ggaaaaacagtgtggatataagctgttcatttatggaagggatgactacgttggggaaga 1321 cattgttgaggtcattaatgaaaacgtaaagaaaagcagaagactgattatcattttagt 1381 cagagaaacatcaggcttcagctggctgggtggttcatctgaagagcaaatagccatgta 1441 taatgctcttgttcaggatggaattaaagttgtcctgcttgagctggagaaaatccaaga 1501 ctatgagaaaatgccagaatcgattaaattcattaagcagaaacatggggctatccgctg 1561 gtcaggggactttacacagggaccacagtctgcaaagacaaggttctggaagaatgtcag 1621 gtaccacatgccagtccagcgacggtcaccttcatctaaacaccagttactgtcaccagc 1681 cactaaggagaaactgcaaagagaggctcacgtgcctctcgggtagcatggagaagttgc 1741 caagagttctttaggtgcctcctgtcttatggcgttgcaggccaggttatgcctcatgct 1801 gacttgcagagttcatggaatgtaactatatcatcctttatccctgaggtcacctggaat 1861 cagattattaagggaataagccatgacgtcaatagcagcccagggcacttcagagtagag 1921 ggcttgggaagatcttttaaaaaggcagtaggcccggtgtggtggctcacgcctataatc 1981 ccagcactttgggaggctgaagtgggtggatcaccagaggtcaggagttcgagaccagcc 2041 cagccaacatggcaaaaccccatctctactaaaaatacaaaaatgagctaggcatggtgg 2101 cacacgcctgtaatcccagctacacctgaggctgaggcaggagaattgcttgaaccgggg 2161 agacggaggttgcagtgagccgagtttgggccactgcactctagcctggcaacagagcaa 2221 gactccgtctcaaaaaaagggcaataaatgccctctctgaatgtttgaactgccaagaaa 2281 aggcatggagacagcgaactagaagaaagggcaagaaggaaatagccaccgtctacagat 2341 ggcttagttaagtcatccacagcccaagggcggggctatgccttgtctggggaccctgta 2401 gagtcactgaccctggagcggctctcctgagaggtgctgcaggcaaagtgagactgacac 2461 ctcactgaggaagggagacatattcttggagaactttccatctgcttgtattttccatac 2521 acatccccagccagaagttagtgtccgaagaccgaattttattttacagagcttgaaaac 2581 tcacttcaatgaacaaagggattctccaggattccaaagttttgaagtcatcttagcttt 2641 ccacaggagggagagaacttaaaaaagcaacagtagcagggaattgatccacttcttaat 2701 gctttcctccctggcatgaccatcctgtcctttgttattatcctgcattttacgtctttg 2761 gaggaacagctccctagtggcttcctccgtctgcaatgtcccttgcacagcccacacatg 2821 aaccatccttcccatgatgccgctcttctgtcatcccgctcctgctgaaacacctcccag 2881 gggctccacctgttcaggagctgaagcccatgctttcccaccagcatgtcactcccagac 2941 cacctccctgccctgtcctccagcttcccctcgctgtcctgctgtgtgaattcccaggtt 3001 ggcctggtggccatgtcgcctgcccccagcactcctctgtctctgctcttgcctgcaccc 3061 ttcctcctcctttgcctaggaggccttctcgcattttctctagctgatcagaattttacc 3121 aaaattcagaacatcctccaattccacagtctctgggagactttccctaagaggcgactt 3181 cctctccagccttctctctctggtcaggcccactgcagagatggtggtgagcacatctgg 3241 gaggctggtctccctccagctggaattgctgctctctgagggagaggctgtggtggctgt 3301 ctctgtccctcactgccttccaggagcaatttgcacatgtaacatagatttatgtaatgc 3361 tttatgtttaaaaacattccccaattatcttatttaatttttgcaattattctaatttta 3421 tatatagagaaagtgacctattttttaaaaaaatcacactctaagttctattgaacctag 3481 gacttgagcctccatttctggcttctagtctggtgttctgagtacttgatttcaggtcaa 3541 taacggtcccccctcactccacactggcacgtttgtgagaagaaatgacattttgctagg 3601 aagtgaccgagtctaggaatgcttttattcaagacaccaaattccaaacttctaaatgtt 3661 ggaattttcaaaaattgtgtttagattttatgaaaaactcttctactttcatctattctt 3721 tccctagaggcaaacatttcttaaaatgtttcattttcattaaaaatgaaagccaaattt 3781 atatgccaccgattgcaggacacaagcacagttttaagagttgtatgaacatggagagga 3841 cttttggtttttatatttctcgtatttaatatgggtgaacaccaacttttatttggaata 3901 ataattttcctcctaaacaaaaacacattgagtttaagtctctgactcttgcctttccac 3961 ctgctttctcctgggcccgctttgcctgcttgaaggaacagtgctgttctggagctgctg 4021 ttccaacagacagggcctagctttcatttgacacacagactacagccagaagcccatgga 4081 gcagggatgtcacgtcttgaaaagcctattagatgttttacaaatttaattttgcagatt 4141 attttagtctgtcatccagaaaatgtgtcagcatgcatagtgctaagaaagcaagccaat 4201 ttggaaacttaggttagtgacaaaattggccagagagtgggggtgatgatgaccaagaat 4261 tacaagtagaatggcagctggaatttaaggagggacaagaatcaatggataagcgtgggt 4321 ggaggaagatccaaacagaaaagtgcaaagttattccccatcttccaagggttgaattct 4381 ggaggaagaagacacattcctagttccccgtgaacttcctttgacttattgtccccacta 4441 aaacaaaacaaaaaacttttaatgccttccacattaattagattttcttgcagttttttt 4501 atggcatttttttaaagatgccctaagtgttgaagaagagtttgcaaatgcaacaaaata 4561 tttaattaccggttgttaaaactggtttagcacaatttatattttccctctcttgccttt 4621 cttatttgcaataaaaggtattgagccattttttaaatgacatttttgataaattatgtt 4681 tgtactagttgatgaaggagttttttttaacctgtttatataattttgcagcagaagcca 4741 aattttttgtatattaaagcaccaaattcatgtacagcatgcatcacggatcaatagact 4801 gtacttattttccaataaaattttcaaactttgtactgttaaaaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant8 (SEQIDNO:111) 1 gtagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggtaaaag 61 acaaggccttctccaagaagaatatgaaagtgttactcagacttatttgtttcatagctc 121 tactgatttcttctctggaggctgataaatgcaaggaacgtgaagaaaaaataattttag 181 tgtcatctgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaacacaaag 241 gcactataacttggtataaagatgacagcaagacacctgtatctacagaacaagcctcca 301 ggattcatcaacacaaagagaaactttggtttgttcctgctaaggtggaggattcaggac 361 attactattgcgtggtaaggattgcaaacctctacttcttgacaatatacactttagtgg 421 agtcaaagataggctcatcgtgatgaatgtggctgaaaagcatagagggaactatacttg 481 tcatgcatcctacacatacttgggcaagcaatatcctattacccgggtaatagaatttat 541 tactctagaggaaaacaaacccacaaggcctgtgattgtgagcccagctaatgagacaat 601 ggaagtagacttgggatcccagatacaattgatctgtaatgtcaccggccagttgagtga 661 cattgcttactggaagtggaatgggtcagtaattgatgaagatgacccagtgctagggga 721 agactattacagtgtggaaaatcctgcaaacaaaagaaggagtaccctcatcacagtgct 781 taatatatcggaaattgaaagtagattttataaacatccatttacctgttttgccaagaa 841 tacacatggtatagatgcagcatatatccagttaatatatccagtcactaatttccagaa 901 gcacatgattggtatatgtgtcacgttgacagtcataattgtgtgttctgttttcatcta 961 taaaatcttcaagattgacattgtgctttggtacagggattcctgctatgattttctccc 1021 aataaaagcttcagatggaaagacctatgacgcatatatactgtatccaaagactgttgg 1081 ggaagggtctacctctgactgtgatatttttgtgtttaaagtcttgcctgaggtcttgga 1141 aaaacagtgtggatataagctgttcatttatggaagggatgactacgttggggaagacat 1201 tgttgaggtcattaatgaaaacgtaaagaaaagcagaagactgattatcattttagtcag 1261 agaaacatcaggcttcagctggctgggtggttcatctgaagagcaaatagccatgtataa 1321 tgctcttgttcaggatggaattaaagttgtcctgcttgagctggagaaaatccaagacta 1381 tgagaaaatgccagaatcgattaaattcattaagcagaaacatggggctatccgctggtc 1441 aggggactttacacagggaccacagtctgcaaagacaaggttctggaagaatgtcaggta 1501 ccacatgccagtccagcgacggtcaccttcatctaaacaccagttactgtcaccagccac 1561 taaggagaaactgcaaagagaggctcacgtgcctctcgggtagcatggagaagttgccaa 1621 gagttctttaggtgcctcctgtcttatggcgttgcaggccaggttatgcctcatgctgac 1681 ttgcagagttcatggaatgtaactatatcatcctttatccctgaggtcacctggaatcag 1741 attattaagggaataagccatgacgtcaatagcagcccagggcacttcagagtagagggc 1801 ttgggaagatcttttaaaaaggcagtaggcccggtgtggtggctcacgcctataatccca 1861 gcactttgggaggctgaagtgggtggatcaccagaggtcaggagttcgagaccagcccag 1921 ccaacatggcaaaaccccatctctactaaaaatacaaaaatgagctaggcatggtggcac 1981 acgcctgtaatcccagctacacctgaggctgaggcaggagaattgcttgaaccggggaga 2041 cggaggttgcagtgagccgagtttgggccactgcactctagcctggcaacagagcaagac 2101 tccgtctcaaaaaaagggcaataaatgccctctctgaatgtttgaactgccaagaaaagg 2161 catggagacagcgaactagaagaaagggcaagaaggaaatagccaccgtctacagatggc 2221 ttagttaagtcatccacagcccaagggcggggctatgccttgtctggggaccctgtagag 2281 tcactgaccctggagcggctctcctgagaggtgctgcaggcaaagtgagactgacacctc 2341 actgaggaagggagacatattcttggagaactttccatctgcttgtattttccatacaca 2401 tccccagccagaagttagtgtccgaagaccgaattttattttacagagcttgaaaactca 2461 cttcaatgaacaaagggattctccaggattccaaagttttgaagtcatcttagctttcca 2521 caggagggagagaacttaaaaaagcaacagtagcagggaattgatccacttcttaatgct 2581 ttcctccctggcatgaccatcctgtcctttgttattatcctgcattttacgtctttggag 2641 gaacagctccctagtggcttcctccgtctgcaatgtcccttgcacagcccacacatgaac 2701 catccttcccatgatgccgctcttctgtcatcccgctcctgctgaaacacctcccagggg 2761 ctccacctgttcaggagctgaagcccatgctttcccaccagcatgtcactcccagaccac 2821 ctccctgccctgtcctccagcttcccctcgctgtcctgctgtgtgaattcccaggttggc 2881 ctggtggccatgtcgcctgcccccagcactcctctgtctctgctcttgcctgcacccttc 2941 ctcctcctttgcctaggaggccttctcgcattttctctagctgatcagaattttaccaaa 3001 attcagaacatcctccaattccacagtctctgggagactttccctaagaggcgacttcct 3061 ctccagccttctctctctggtcaggcccactgcagagatggtggtgagcacatctgggag 3121 gctggtctccctccagctggaattgctgctctctgagggagaggctgtggtggctgtctc 3181 tgtccctcactgccttccaggagcaatttgcacatgtaacatagatttatgtaatgcttt 3241 atgtttaaaaacattccccaattatcttatttaatttttgcaattattctaattttatat 3301 atagagaaagtgacctattttttaaaaaaatcacactctaagttctattgaacctaggac 3361 ttgagcctccatttctggcttctagtctggtgttctgagtacttgatttcaggtcaataa 3421 cggtcccccctcactccacactggcacgtttgtgagaagaaatgacattttgctaggaag 3481 tgaccgagtctaggaatgcttttattcaagacaccaaattccaaacttctaaatgttgga 3541 attttcaaaaattgtgtttagattttatgaaaaactcttctactttcatctattctttcc 3601 ctagaggcaaacatttcttaaaatgtttcattttcattaaaaatgaaagccaaatttata 3661 tgccaccgattgcaggacacaagcacagttttaagagttgtatgaacatggagaggactt 3721 ttggtttttatatttctcgtatttaatatgggtgaacaccaacttttatttggaataata 3781 attttcctcctaaacaaaaacacattgagtttaagtctctgactcttgcctttccacctg 3841 ctttctcctgggcccgctttgcctgcttgaaggaacagtgctgttctggagctgctgttc 3901 caacagacagggcctagctttcatttgacacacagactacagccagaagcccatggagca 3961 gggatgtcacgtcttgaaaagcctattagatgttttacaaatttaattttgcagattatt 4021 ttagtctgtcatccagaaaatgtgtcagcatgcatagtgctaagaaagcaagccaatttg 4081 gaaacttaggttagtgacaaaattggccagagagtgggggtgatgatgaccaagaattac 4141 aagtagaatggcagctggaatttaaggagggacaagaatcaatggataagcgtgggtgga 4201 ggaagatccaaacagaaaagtgcaaagttattccccatcttccaagggttgaattctgga 4261 ggaagaagacacattcctagttccccgtgaacttcctttgacttattgtccccactaaaa 4321 caaaacaaaaaacttttaatgccttccacattaattagattttcttgcagtttttttatg 4381 gcatttttttaaagatgccctaagtgttgaagaagagtttgcaaatgcaacaaaatattt 4441 aattaccggttgttaaaactggtttagcacaatttatattttccctctcttgcctactt 4501 atttgcaataaaaggtattgagccattttttaaatgacatttttgataaattatgtttgt 4561 actagttgatgaaggagttttttttaacctgtttatataattttgcagcagaagccaaat 4621 tttttgtatattaaagcaccaaattcatgtacagcatgcatcacggatcaatagactgta 4681 cttattttccaataaaattttcaaactttgtactgttaaaaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant9 (SEQIDNO:112) 1 gtagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggtaaaag 61 acaaggccttctccaagaagaatatgaaagtgttactcagacttatttgtttcatagctc 121 tactgatttcttctctggaggctgataaatgcaaggaacgtgaagaaaaaataattttag 181 tgtcatctgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaacacaaag 241 gcactataacttggtataaagatgacagcaagacacctgtatctacagaacaagcctcca 301 ggattcatcaacacaaagagaaactttggtttgttcctgctaaggtggaggattcaggac 361 attactattgcgtggtaagaaattcatcttactgcctcagaattaaaataagtgcaaaat 421 ttgtggagaatgagcctaacttatgttataatgcacaagccatatttaagcagaaactac 481 ccgttgcaggagacggaggacttgtgtgcccttatatggagttttttaaaaatgaaaata 541 atgagttacctaaattacagtggtataagaggaaaacaaacccacaaggcctgtgattgt 601 gagcccagctaatgagacaatggaagtagacttgggatcccagatacaattgatctgtaa 661 tgtcaccggccagttgagtgacattgcttactggaagtggaatgggtcagtaattgatga 721 agatgacccagtgctaggggaagactattacagtgtggaaaatcctgcaaacaaaagaag 781 gagtaccctcatcacagtgcttaatatatcggaaattgaaagtagattttataaacatcc 841 atttacctgttttgccaagaatacacatggtatagatgcagcatatatccagttaatata 901 tccagtcactaatttccagaagcacatgattggtatatgtgtcacgttgacagtcataat 961 tgtgtgttctgttttcatctataaaatcttcaagattgacattgtgctttggtacaggga 1021 ttcctgctatgattttctcccaataaaagcttcagatggaaagacctatgacgcatatat 1081 actgtatccaaagactgttggggaagggtctacctctgactgtgatatttttgtgtttaa 1141 agtcttgcctgaggtcttggaaaaacagtgtggatataagctgttcatttatggaaggga 1201 tgactacgttggggaagacattgttgaggtcattaatgaaaacgtaaagaaaagcagaag 1261 actgattatcattttagtcagagaaacatcaggcttcagctggctgggtggttcatctga 1321 agagcaaatagccatgtataatgctcttgttcaggatggaattaaagttgtcctgcttga 1381 gctggagaaaatccaagactatgagaaaatgccagaatcgattaaattcattaagcagaa 1441 acatggggctatccgctggtcaggggactttacacagggaccacagtctgcaaagacaag 1501 gttctggaagaatgtcaggtaccacatgccagtccagcgacggtcaccttcatctaaaca 1561 ccagttactgtcaccagccactaaggagaaactgcaaagagaggctcacgtgcctctcgg 1621 gtagcatggagaagttgccaagagttctttaggtgcctcctgtcttatggcgttgcaggc 1681 caggttatgcctcatgctgacttgcagagttcatggaatgtaactatatcatccatatc 1741 cctgaggtcacctggaatcagattattaagggaataagccatgacgtcaatagcagccca 1801 gggcacttcagagtagagggcttgggaagatcttttaaaaaggcagtaggcccggtgtgg 1861 tggctcacgcctataatcccagcactttgggaggctgaagtgggtggatcaccagaggtc 1921 aggagttcgagaccagcccagccaacatggcaaaaccccatctctactaaaaatacaaaa 1981 atgagctaggcatggtggcacacgcctgtaatcccagctacacctgaggctgaggcagga 2041 gaattgcttgaaccggggagacggaggttgcagtgagccgagtttgggccactgcactct 2101 agcctggcaacagagcaagactccgtctcaaaaaaagggcaataaatgccctctctgaat 2161 gtttgaactgccaagaaaaggcatggagacagcgaactagaagaaagggcaagaaggaaa 2221 tagccaccgtctacagatggcttagttaagtcatccacagcccaagggcggggctatgcc 2281 ttgtctggggaccctgtagagtcactgaccctggagcggctctcctgagaggtgctgcag 2341 gcaaagtgagactgacacctcactgaggaagggagacatattcttggagaactttccatc 2401 tgcttgtattttccatacacatccccagccagaagttagtgtccgaagaccgaattttat 2461 tttacagagcttgaaaactcacttcaatgaacaaagggattctccaggattccaaagttt 2521 tgaagtcatcttagctttccacaggagggagagaacttaaaaaagcaacagtagcaggga 2581 attgatccacttcttaatgctttcctccctggcatgaccatcctgtcctttgttattatc 2641 ctgcattttacgtctttggaggaacagctccctagtggcttcctccgtctgcaatgtccc 2701 ttgcacagcccacacatgaaccatccttcccatgatgccgctcttctgtcatcccgctcc 2761 tgctgaaacacctcccaggggctccacctgttcaggagctgaagcccatgctttcccacc 2821 agcatgtcactcccagaccacctccctgccctgtcctccagcttcccctcgctgtcctgc 2881 tgtgtgaattcccaggttggcctggtggccatgtcgcctgcccccagcactcctctgtct 2941 ctgctcttgcctgcacccttcctcctcctttgcctaggaggccttctcgcattttctcta 3001 gctgatcagaattttaccaaaattcagaacatcctccaattccacagtctctgggagact 3061 ttccctaagaggcgacttcctctccagccttctctctctggtcaggcccactgcagagat 3121 ggtggtgagcacatctgggaggctggtctccctccagctggaattgctgctctctgaggg 3181 agaggctgtggtggctgtctctgtccctcactgccttccaggagcaatttgcacatgtaa 3241 catagatttatgtaatgctttatgtttaaaaacattccccaattatcttatttaattttt 3301 gcaattattctaattttatatatagagaaagtgacctattttttaaaaaaatcacactct 3361 aagttctattgaacctaggacttgagcctccatttctggcttctagtctggtgttctgag 3421 tacttgatttcaggtcaataacggtcccccctcactccacactggcacgtttgtgagaag 3481 aaatgacattttgctaggaagtgaccgagtctaggaatgcttttattcaagacaccaaat 3541 tccaaacttctaaatgttggaattttcaaaaattgtgtttagattttatgaaaaactctt 3601 ctactttcatctattctttccctagaggcaaacatttcttaaaatgtttcattttcatta 3661 aaaatgaaagccaaatttatatgccaccgattgcaggacacaagcacagttttaagagtt 3721 gtatgaacatggagaggacttttggtttttatatttctcgtatttaatatgggtgaacac 3781 caacttttatttggaataataattttcctcctaaacaaaaacacattgagtttaagtctc 3841 tgactcttgcctttccacctgctttctcctgggcccgctttgcctgcttgaaggaacagt 3901 gctgttctggagctgctgttccaacagacagggcctagctttcatttgacacacagacta 3961 cagccagaagcccatggagcagggatgtcacgtcttgaaaagcctattagatgttttaca 4021 aatttaattttgcagattattttagtctgtcatccagaaaatgtgtcagcatgcatagtg 4081 ctaagaaagcaagccaatttggaaacttaggttagtgacaaaattggccagagagtgggg 4141 gtgatgatgaccaagaattacaagtagaatggcagctggaatttaaggagggacaagaat 4201 caatggataagcgtgggtggaggaagatccaaacagaaaagtgcaaagttattccccatc 4261 ttccaagggttgaattctggaggaagaagacacattcctagttccccgtgaacttccttt 4321 gacttattgtccccactaaaacaaaacaaaaaacttttaatgccttccacattaattaga 4381 ttttcttgcagtttttttatggcatttttttaaagatgccctaagtgttgaagaagagtt 4441 tgcaaatgcaacaaaatatttaattaccggttgttaaaactggtttagcacaatttatat 4501 tttccctctcttgcctttcttatttgcaataaaaggtattgagccattttttaaatgaca 4561 tttttgataaattatgtttgtactagttgatgaaggagttttttttaacctgtttatata 4621 attttgcagcagaagccaaattttttgtatattaaagcaccaaattcatgtacagcatgc 4681 atcacggatcaatagactgtacttattttccaataaaattttcaaactttgtactgttaa 4741 aaaaaaaaaaaaaaaa HumanIL-1R1mRNAVariant10 (SEQIDNO:113) 1 attaaagccctaagaggctgtgacacagccatctccaaaaccccactttctccttccttt 61 gagcctccgtaccagctggggcgtccggcaagatgtgagttgtcactctgctgcggcaca 121 gacctgaattaacaactctagctagggctgacttcaaaaagcactttcgttttttaataa 181 ccaacatcagctcagcaggcttcatttgggaaaagaaaccttgtcggattaccccgacat 241 tctccacctcctgggaggccagccattcccaaatgccccaaggatgaagaacggagacgg 301 tagacgcaccctctgaagatggtgactccctcctgagaagctggaccccttggtaaaaga 361 caaggccttctccaagaagaatatgaaagtgttactcagacttatttgtttcatagctct 421 actgatttcttctctggaggctgataaatgcaaggaacgtgaagaaaaaataattttagt 481 gtcatctgcaaatgaaattgatgttcgtccctgtcctcttaacccaaatgaacacaaagg 541 cactataacttggtataaagatgacagcaagacacctgtatctacagaacaagcctccag 601 gattcatcaacacaaagagaaactttggtttgttcctgctaaggtggaggattcaggaca 661 ttactattgcgtggtaagaaattcatcttactgcctcagaattaaaataagtgcaaaatt 721 tgtggagaatgagcctaacttatgttataatgcacaagccatatttaagcagaaactacc 781 cgttgcaggagacggaggacttgtgtgcccttatatggagttttttaaaaatgaaaataa 841 tgagttacctaaattacagtggtataaggtaattttattttaaatatgacatttcacttt 901 tccagaaaataaaatagttccctggacaatagaaaaaaaaaaaaaaaaaa HumanIL1RAPmRNAVariant1 (SEQIDNO:114) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaaggcatcgtcatg 181 tgatcatcacctaagaactagaacatcagcaggccctagaagcctcactcttgcccctcc 241 ctttaatatctcaaaggatgacacttctgtggtgtgtagtgagtctctacttttatggaa 301 tcctgcaaagtgatgcctcagaacgctgcgatgactggggactagacaccatgaggcaaa 361 tccaagtgtttgaagatgagccagctcgcatcaagtgcccactctttgaacacttcttga 421 aattcaactacagcacagcccattcagctggccttactctgatctggtattggactaggc 481 aggaccgggaccttgaggagccaattaacttccgcctccccgagaaccgcattagtaagg 541 agaaagatgtgctgtggttccggcccactctcctcaatgacactggcaactatacctgca 601 tgttaaggaacactacatattgcagcaaagttgcatttcccttggaagttgttcaaaaag 661 acagctgtttcaattcccccatgaaactcccagtgcataaactgtatatagaatatggca 721 ttcagaggatcacttgtccaaatgtagatggatattttccttccagtgtcaaaccgacta 781 tcacttggtatatgggctgttataaaatacagaattttaataatgtaatacccgaaggta 841 tgaacttgagtttcctcattgccttaatttcaaataatggaaattacacatgtgttgtta 901 catatccagaaaatggacgtacgtttcatctcaccaggactctgactgtaaaggtagtag 961 gctctccaaaaaatgcagtgccccctgtgatccattcacctaatgatcatgtggtctatg 1021 agaaagaaccaggagaggagctactcattccctgtacggtctattttagttttctgatgg 1081 attctcgcaatgaggtttggtggaccattgatggaaaaaaacctgatgacatcactattg 1141 atgtcaccattaacgaaagtataagtcatagtagaacagaagatgaaacaagaactcaga 1201 ttttgagcatcaagaaagttacctctgaggatctcaagcgcagctatgtctgtcatgcta 1261 gaagtgccaaaggcgaagttgccaaagcagccaaggtgaagcagaaagtgccagctccaa 1321 gatacacagtggaactggcttgtggttttggagccacagtcctgctagtggtgattctca 1381 ttgttgtttaccatgtttactggctagagatggtcctattttaccgggctcattttggaa 1441 cagatgaaaccattttagatggaaaagagtatgatatttatgtatcctatgcaaggaatg 1501 cggaagaagaagaatttgtattactgaccctccgtggagttttggagaatgaatttggat 1561 acaagctgtgcatctttgaccgagacagtctgcctgggggaattgtcacagatgagactt 1621 tgagcttcattcagaaaagcagacgcctcctggttgttctaagccccaactacgtgctcc 1681 agggaacccaagccctcctggagctcaaggctggcctagaaaatatggcctctcggggca 1741 acatcaacgtcattttagtacagtacaaagctgtgaaggaaacgaaggtgaaagagctga 1801 agagggctaagacggtgctcacggtcattaaatggaaaggggaaaaatccaagtatccac 1861 agggcaggttctggaagcagctgcaggtggccatgccagtgaagaaaagtcccaggcggt 1921 ctagcagtgatgagcagggcctctcgtattcatctttgaaaaatgtatgaaaggaataat 1981 gaaaagggtaaaaagaacaaggggtgctccaggaagaaagagtccccccagtcttcattc 2041 gcagtttatggtttcataggcaaaaataatggtctaagcctcccaatagggataaattta 2101 gggtgactgtgtggctgactattctgcttcctcaggcaacactaaagtttagaaagatat 2161 catcaacgttctgtcaccagtctctgatgccactatgttctttgcaggcaaagacttgtt 2221 caatgcgaatttccccttctacattgtctatccctgtttttatatgtctccattcttttt 2281 aaaatcttaacatatggagcagcctttcctatgaatttaaatatgcctttaaaataagtc 2341 actgttgacagggtcatgagtttccgagtatagttttctttttatcttatttttactcgt 2401 ccgttgaaaagataatcaaggcctacattttagctgaggataatgaacttttttcctcat 2461 tcggctgtataatacataaccacagcaagactgacatccacttaggatgatacaaagcag 2521 tgtaactgaaaatgtttcttttaattgatttaaaggacttgtcttctataccacccttgt 2581 cctcatctcaggtaatttatgaaatctatgtaaacttgaaaaatatttcttaatttttgt 2641 ttttgctccagtcaattcctgattatccacaggtcaacccacattttttcattccttctc 2701 cctatctgcttatatcgcattgctcatttagagtttgcaggaggctccatactaggttca 2761 gtctgaaagaaatctcctaatggtgctatagagagggaggtaacagaaagactcttttag 2821 ggcatttttctgactcatgaaaagagcacagaaaaggatgtttggcaatttgtcttttaa 2881 gtcttaaccttgctaatgtgaatactgggaaagtgattttttctcactcgtttttgttgc 2941 tccattgtaaagggcggaggtcagtcttagtggccttgagagttgcttttggcattaata 3001 ttctaagagaattaactgtatttcctgtcacctattcactagtgcaggaaatatacttgc 3061 tccaaataagtcagtatgagaagtcactgtcaatgaaagttgttttgtttgttttcagta 3121 atattttgctgtttttaagacttggaaaactaagtgcagagtttacagagtggtaaatat 3181 ctatgttacatgtagattatacatatatatacacacgtgtatatgagatatatatcttat 3241 atctccacaaacacaaattatatatatacatatccacacacatacattacatatatctgt 3301 gtatataaatccacatgcacatgaaatatatatatatatataatttgtgtgtgtgtatgt 3361 gtatgtatatgactttaaatagctatgggtacaatattaaaaaccactggaactcttgtc 3421 cagtttttaaattatgtttttactggaatgtttttgtgtcagtgttttctgtacatatta 3481 tttgttaattcacagctcacagagtgatagttgtcatagttcttgccttccctaagttta 3541 tataaataacttaagtattgctacagtttatctaggttgcagtggcatctgctgtgcaca 3601 gagcttccatggtcactgctaagcagtagccagccatcgggcattaattgatttcctact 3661 atattcccagcagacacatttagaaactaagctatgttaacctcagtgctcaactatttg 3721 aactgttgagtgataaaggaaacaaatataactgtaaatgaatcttggtatcctgtgaaa 3781 cagaataattcgtaatttaagaaagcccttatcccggtaacatgaatgttgatgaacaaa 3841 tgtaaaattatatcctatatttaagtacccataataaatcatttccctctataagtgtta 3901 ttgattattttaaattgaaaaaagtttcacttggatgaaaaaagtagaaaagtaggtcat 3961 tcttggatctacttttttttagccttattaatatttttccctattagaaaccacaattac 4021 tccctctattaacccttcacttactagaccagaaaagaacttattccagataagctttga 4081 atatcaattcttacataaactttaggcaaacagggaatagtctagtcaccaaaggaccat 4141 tctcttgccaatgctgcattccttttgcacttttggattccatatttatcccaaatgctg 4201 ttgggcacccctagaaataccttgatgttttttctatttatatgcctgcctttggtactt 4261 aattttacaaatgctgtaatataaagcatatcaagtttatgtgatacgtatcattgcaag 4321 agaatttgtttcaagatttttttttaatgttccagaagatggccaatagagaacattcaa 4381 gggaaatggggaaacataatttagagaacaagaacaaaccatgtctcaaatttttttaaa 4441 aaaaattaatggttttaaatatatgctatagggacgttccatgcccaggttaacaaagaa 4501 ctgtgatatatagagtgtctaattacaaaatcatatacgatttatttaattctcttctgt 4561 attgtaacttagatgattcccaaggactctaataaaaaatcacttcattgtatttggaaa 4621 caaaaacatcattcattaattacttattttctttccataggttttaatattttgagagtg 4681 tcttttttatttcattcatgaacttttgtatttttcatttttcatttgatttgtaaattt 4741 acttatgttaaaaataaaccatttattttcagctttgaattttaaaaaaaaaaaaaaaaa 4801 a HumanIL1RAPmRNAVariant2 (SEQIDNO:115) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaaggcatcgtcatg 181 tgatcatcacctaagaactagaacatcagcaggccctagaagcctcactcttgcccctcc 241 ctttaatatctcaaaggatgacacttctgtggtgtgtagtgagtctctacttttatggaa 301 tcctgcaaagtgatgcctcagaacgctgcgatgactggggactagacaccatgaggcaaa 361 tccaagtgtttgaagatgagccagctcgcatcaagtgcccactctttgaacacttcttga 421 aattcaactacagcacagcccattcagctggccttactctgatctggtattggactaggc 481 aggaccgggaccttgaggagccaattaacttccgcctccccgagaaccgcattagtaagg 541 agaaagatgtgctgtggttccggcccactctcctcaatgacactggcaactatacctgca 601 tgttaaggaacactacatattgcagcaaagttgcatttcccttggaagttgttcaaaaag 661 acagctgtttcaattcccccatgaaactcccagtgcataaactgtatatagaatatggca 721 ttcagaggatcacttgtccaaatgtagatggatattttccttccagtgtcaaaccgacta 781 tcacttggtatatgggctgttataaaatacagaattttaataatgtaatacccgaaggta 841 tgaacttgagtttcctcattgccttaatttcaaataatggaaattacacatgtgttgtta 901 catatccagaaaatggacgtacgtttcatctcaccaggactctgactgtaaaggtagtag 961 gctctccaaaaaatgcagtgccccctgtgatccattcacctaatgatcatgtggtctatg 1021 agaaagaaccaggagaggagctactcattccctgtacggtctattttagttttctgatgg 1081 attctcgcaatgaggtttggtggaccattgatggaaaaaaacctgatgacatcactattg 1141 atgtcaccattaacgaaagtataagtcatagtagaacagaagatgaaacaagaactcaga 1201 ttttgagcatcaagaaagttacctctgaggatctcaagcgcagctatgtctgtcatgcta 1261 gaagtgccaaaggcgaagttgccaaagcagccaaggtgaagcagaaaggtaatagatgcg 1321 gtcagtgatgaatctctcagctccaaattaacattgtggtgaataaggacaaaaggagag 1381 attgagaacaagagagctccagcacctagcccgacggcatctaacccatagtaatgaatc 1441 aaacttaaatgaaaaatatgaaagttttcatctatgtaagatactcaaaatattgtttct 1501 gatattgttagtaccgtaatgcccaaatgtagctaaaaaaatcgacgtgagtacagtgag 1561 acacaattttgtgtctgtacaattatgaaaaattaaaaacaaagaaaatattcaaagcta 1621 ccaaagatagaaaaaactggtagagccacatattgttggtgaattattaagaccctttta 1681 aaaatcattcatggtagacttcaagagtcataaaaaagattgcatcatctgacctaagac 1741 tttcggaatttttcctgaacaaataacagaaagggaattatataccttttaatattatta 1801 gaagcattatctgtagttgtaaaacattattaatagcagccatccaattgtatgcaacta 1861 attaaggtattgaatgtttattttccaaaaatgcataattataatattattttaaacact 1921 atgtatcaatatttaagcaggtttataatataccagcagccacaattgctaaaatgaaaa 1981 tcatttaaattatgattttaaatggtataaacatgatttctatgttgatagtactatatt 2041 attctacaataaatggaaattataaagccttcttgtcagaagtgctgctcctaaaaaaaa 2101 aaaaaaaaaaaaaa HumanIL1RAPmRNAVariant3 (SEQIDNO:116) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaagggctttgacct 181 gaagcttgaaattgagtttgggacaataatgtgtctcatggggaattgcatggactcctt 241 atcataagccaaatgctgaggtaaagctgcggaattgagtcgtcctccaagaagggagag 301 aaaatgatgtcttgtgacatttccagataactggcatcgtcatgtgatcatcacctaaga 361 actagaacatcagcaggccctagaagcctcactcttgcccctccctttaatatctcaaag 421 gatgacacttctgtggtgtgtagtgagtctctacttttatggaatcctgcaaagtgatgc 481 ctcagaacgctgcgatgactggggactagacaccatgaggcaaatccaagtgtttgaaga 541 tgagccagctcgcatcaagtgcccactctttgaacacttcttgaaattcaactacagcac 601 agcccattcagctggccttactctgatctggtattggactaggcaggaccgggaccttga 661 ggagccaattaacttccgcctccccgagaaccgcattagtaaggagaaagatgtgctgtg 721 gttccggcccactctcctcaatgacactggcaactatacctgcatgttaaggaacactac 781 atattgcagcaaagttgcatttcccttggaagttgttcaaaaagacagctgtttcaattc 841 ccccatgaaactcccagtgcataaactgtatatagaatatggcattcagaggatcacttg 901 tccaaatgtagatggatattttccttccagtgtcaaaccgactatcacttggtatatggg 961 ctgttataaaatacagaattttaataatgtaatacccgaaggtatgaacttgagtttcct 1021 cattgccttaatttcaaataatggaaattacacatgtgttgttacatatccagaaaatgg 1081 acgtacgtttcatctcaccaggactctgactgtaaaggtagtaggctctccaaaaaatgc 1141 agtgccccctgtgatccattcacctaatgatcatgtggtctatgagaaagaaccaggaga 1201 ggagctactcattccctgtacggtctattttagttttctgatggattctcgcaatgaggt 1261 ttggtggaccattgatggaaaaaaacctgatgacatcactattgatgtcaccattaacga 1321 aagtataagtcatagtagaacagaagatgaaacaagaactcagattttgagcatcaagaa 1381 agttacctctgaggatctcaagcgcagctatgtctgtcatgctagaagtgccaaaggcga 1441 agttgccaaagcagccaaggtgaagcagaaagtgccagctccaagatacacagtggaact 1501 ggcttgtggttttggagccacagtcctgctagtggtgattctcattgttgtttaccatgt 1561 ttactggctagagatggtcctattttaccgggctcattttggaacagatgaaaccatttt 1621 agatggaaaagagtatgatatttatgtatcctatgcaaggaatgcggaagaagaagaatt 1681 tgtattactgaccctccgtggagttttggagaatgaatttggatacaagctgtgcatctt 1741 tgaccgagacagtctgcctgggggaattgtcacagatgagactttgagcttcattcagaa 1801 aagcagacgcctcctggttgttctaagccccaactacgtgctccagggaacccaagccct 1861 cctggagctcaaggctggcctagaaaatatggcctctcggggcaacatcaacgtcatttt 1921 agtacagtacaaagctgtgaaggaaacgaaggtgaaagagctgaagagggctaagacggt 1981 gctcacggtcattaaatggaaaggggaaaaatccaagtatccacagggcaggttctggaa 2041 gcagctgcaggtggccatgccagtgaagaaaagtcccaggcggtctagcagtgatgagca 2101 gggcctctcgtattcatctttgaaaaatgtatgaaaggaataatgaaaagggtaaaaaga 2161 acaaggggtgctccaggaagaaagagtccccccagtcttcattcgcagtttatggtttca 2221 taggcaaaaataatggtctaagcctcccaatagggataaatttagggtgactgtgtggct 2281 gactattctgcttcctcaggcaacactaaagtttagaaagatatcatcaacgttctgtca 2341 ccagtctctgatgccactatgttctttgcaggcaaagacttgttcaatgcgaatttcccc 2401 ttctacattgtctatccctgtttttatatgtctccattctttttaaaatcttaacatatg 2461 gagcagcctttcctatgaatttaaatatgcctttaaaataagtcactgttgacagggtca 2521 tgagtttccgagtatagttttctttttatcttatttttactcgtccgttgaaaagataat 2581 caaggcctacattttagctgaggataatgaacttttttcctcattcggctgtataataca 2641 taaccacagcaagactgacatccacttaggatgatacaaagcagtgtaactgaaaatgtt 2701 tcttttaattgatttaaaggacttgtcttctataccacccttgtcctcatctcaggtaat 2761 ttatgaaatctatgtaaacttgaaaaatatttcttaatttttgtttttgctccagtcaat 2821 tcctgattatccacaggtcaacccacattttttcattccttctccctatctgcttatatc 2881 gcattgctcatttagagtttgcaggaggctccatactaggttcagtctgaaagaaatctc 2941 ctaatggtgctatagagagggaggtaacagaaagactcttttagggcatttttctgactc 3001 atgaaaagagcacagaaaaggatgtttggcaatttgtcttttaagtcttaaccttgctaa 3061 tgtgaatactgggaaagtgattttttctcactcgtttttgttgctccattgtaaagggcg 3121 gaggtcagtcttagtggccttgagagttgcttttggcattaatattctaagagaattaac 3181 tgtatttcctgtcacctattcactagtgcaggaaatatacttgctccaaataagtcagta 3241 tgagaagtcactgtcaatgaaagttgttttgtttgttttcagtaatattttgctgttttt 3301 aagacttggaaaactaagtgcagagtttacagagtggtaaatatctatgttacatgtaga 3361 ttatacatatatatacacacgtgtatatgagatatatatcttatatctccacaaacacaa 3421 attatatatatacatatccacacacatacattacatatatctgtgtatataaatccacat 3481 gcacatgaaatatatatatatatataatttgtgtgtgtgtatgtgtatgtatatgacttt 3541 aaatagctatgggtacaatattaaaaaccactggaactcttgtccagtttttaaattatg 3601 tttttactggaatgtttttgtgtcagtgttttctgtacatattatttgttaattcacagc 3661 tcacagagtgatagttgtcatagttcttgccttccctaagtttatataaataacttaagt 3721 attgctacagtttatctaggttgcagtggcatctgctgtgcacagagcttccatggtcac 3781 tgctaagcagtagccagccatcgggcattaattgatttcctactatattcccagcagaca 3841 catttagaaactaagctatgttaacctcagtgctcaactatttgaactgttgagtgataa 3901 aggaaacaaatataactgtaaatgaatcttggtatcctgtgaaacagaataattcgtaat 3961 ttaagaaagcccttatcccggtaacatgaatgttgatgaacaaatgtaaaattatatcct 4021 atatttaagtacccataataaatcatttccctctataagtgttattgattattttaaatt 4081 gaaaaaagtttcacttggatgaaaaaagtagaaaagtaggtcattcttggatctactttt 4141 ttttagccttattaatatttttccctattagaaaccacaattactccctctattaaccct 4201 tcacttactagaccagaaaagaacttattccagataagctttgaatatcaattcttacat 4261 aaactttaggcaaacagggaatagtctagtcaccaaaggaccattctcttgccaatgctg 4321 cattccttttgcacttttggattccatatttatcccaaatgctgttgggcacccctagaa 4381 ataccttgatgttttttctatttatatgcctgcctttggtacttaattttacaaatgctg 4441 taatataaagcatatcaagtttatgtgatacgtatcattgcaagagaatttgtttcaaga 4501 tttttttttaatgttccagaagatggccaatagagaacattcaagggaaatggggaaaca 4561 taatttagagaacaagaacaaaccatgtctcaaatttttttaaaaaaaattaatggtttt 4621 aaatatatgctatagggacgttccatgcccaggttaacaaagaactgtgatatatagagt 4681 gtctaattacaaaatcatatacgatttatttaattctcttctgtattgtaacttagatga 4741 ttcccaaggactctaataaaaaatcacttcattgtatttggaaacaaaaacatcattcat 4801 taattacttattttctttccataggttttaatattttgagagtgtcttttttatttcatt 4861 catgaacttttgtatttttcatttttcatttgatttgtaaatttacttatgttaaaaata 4921 aaccatttattttcagctttgaattttaaaaaaaaaaaaaaaaaa HumanIL1RAPmRNAVariant4 (SEQIDNO:117) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaaggatgacacttc 181 tgtggtgtgtagtgagtctctacttttatggaatcctgcaaagtgatgcctcagaacgct 241 gcgatgactggggactagacaccatgaggcaaatccaagtgtttgaagatgagccagctc 301 gcatcaagtgcccactctttgaacacttcttgaaattcaactacagcacagcccattcag 361 ctggccttactctgatctggtattggactaggcaggaccgggaccttgaggagccaatta 421 acttccgcctccccgagaaccgcattagtaaggagaaagatgtgctgtggttccggccca 481 ctctcctcaatgacactggcaactatacctgcatgttaaggaacactacatattgcagca 541 aagttgcatttcccttggaagttgttcaaaaagacagctgtttcaattcccccatgaaac 601 tcccagtgcataaactgtatatagaatatggcattcagaggatcacttgtccaaatgtag 661 atggatattttccttccagtgtcaaaccgactatcacttggtatatgggctgttataaaa 721 tacagaattttaataatgtaatacccgaaggtatgaacttgagtttcctcattgccttaa 781 tttcaaataatggaaattacacatgtgttgttacatatccagaaaatggacgtacgtttc 841 atctcaccaggactctgactgtaaaggtagtaggctctccaaaaaatgcagtgccccctg 901 tgatccattcacctaatgatcatgtggtctatgagaaagaaccaggagaggagctactca 961 ttccctgtacggtctattttagttttctgatggattctcgcaatgaggtttggtggacca 1021 ttgatggaaaaaaacctgatgacatcactattgatgtcaccattaacgaaagtataagtc 1081 atagtagaacagaagatgaaacaagaactcagattttgagcatcaagaaagttacctctg 1141 aggatctcaagcgcagctatgtctgtcatgctagaagtgccaaaggcgaagttgccaaag 1201 cagccaaggtgaagcagaaagtgccagctccaagatacacagtggaactggcttgtggtt 1261 ttggagccacagtcctgctagtggtgattctcattgttgtttaccatgtttactggctag 1321 agatggtcctattttaccgggctcattttggaacagatgaaaccattttagatggaaaag 1381 agtatgatatttatgtatcctatgcaaggaatgcggaagaagaagaatttgtattactga 1441 ccctccgtggagttttggagaatgaatttggatacaagctgtgcatctttgaccgagaca 1501 gtctgcctgggggaattgtcacagatgagactttgagcttcattcagaaaagcagacgcc 1561 tcctggttgttctaagccccaactacgtgctccagggaacccaagccctcctggagctca 1621 aggctggcctagaaaatatggcctctcggggcaacatcaacgtcattttagtacagtaca 1681 aagctgtgaaggaaacgaaggtgaaagagctgaagagggctaagacggtgctcacggtca 1741 ttaaatggaaaggggaaaaatccaagtatccacagggcaggttctggaagcagctgcagg 1801 tggccatgccagtgaagaaaagtcccaggcggtctagcagtgatgagcagggcctctcgt 1861 attcatctttgaaaaatgtatgaaaggaataatgaaaagggtaaaaagaacaaggggtgc 1921 tccaggaagaaagagtccccccagtcttcattcgcagtttatggtttcataggcaaaaat 1981 aatggtctaagcctcccaatagggataaatttagggtgactgtgtggctgactattctgc 2041 ttcctcaggcaacactaaagtttagaaagatatcatcaacgttctgtcaccagtctctga 2101 tgccactatgttctttgcaggcaaagacttgttcaatgcgaatttccccttctacattgt 2161 ctatccctgtttttatatgtctccattctttttaaaatcttaacatatggagcagccttt 2221 cctatgaatttaaatatgcctttaaaataagtcactgttgacagggtcatgagtttccga 2281 gtatagttttctttttatcttatttttactcgtccgttgaaaagataatcaaggcctaca 2341 ttttagctgaggataatgaacttttttcctcattcggctgtataatacataaccacagca 2401 agactgacatccacttaggatgatacaaagcagtgtaactgaaaatgtttcttttaattg 2461 atttaaaggacttgtcttctataccacccttgtcctcatctcaggtaatttatgaaatct 2521 atgtaaacttgaaaaatatttcttaatttttgtttttgctccagtcaattcctgattatc 2581 cacaggtcaacccacattttttcattccttctccctatctgcttatatcgcattgctcat 2641 ttagagtttgcaggaggctccatactaggttcagtctgaaagaaatctcctaatggtgct 2701 atagagagggaggtaacagaaagactcttttagggcatttttctgactcatgaaaagagc 2761 acagaaaaggatgtttggcaatttgtcttttaagtcttaaccttgctaatgtgaatactg 2821 ggaaagtgattttttctcactcgtttttgttgctccattgtaaagggcggaggtcagtct 2881 tagtggccttgagagttgcttttggcattaatattctaagagaattaactgtatttcctg 2941 tcacctattcactagtgcaggaaatatacttgctccaaataagtcagtatgagaagtcac 3001 tgtcaatgaaagttgttttgtttgttttcagtaatattttgctgtttttaagacttggaa 3061 aactaagtgcagagtttacagagtggtaaatatctatgttacatgtagattatacatata 3121 tatacacacgtgtatatgagatatatatcttatatctccacaaacacaaattatatatat 3181 acatatccacacacatacattacatatatctgtgtatataaatccacatgcacatgaaat 3241 atatatatatatataatttgtgtgtgtgtatgtgtatgtatatgactttaaatagctatg 3301 ggtacaatattaaaaaccactggaactcttgtccagtttttaaattatgtttttactgga 3361 atgtttttgtgtcagtgttttctgtacatattatttgttaattcacagctcacagagtga 3421 tagttgtcatagttcttgccttccctaagtttatataaataacttaagtattgctacagt 3481 ttatctaggttgcagtggcatctgctgtgcacagagcttccatggtcactgctaagcagt 3541 agccagccatcgggcattaattgatttcctactatattcccagcagacacatttagaaac 3601 taagctatgttaacctcagtgctcaactatttgaactgttgagtgataaaggaaacaaat 3661 ataactgtaaatgaatcttggtatcctgtgaaacagaataattcgtaatttaagaaagcc 3721 cttatcccggtaacatgaatgttgatgaacaaatgtaaaattatatcctatatttaagta 3781 cccataataaatcatttccctctataagtgttattgattattttaaattgaaaaaagttt 3841 cacttggatgaaaaaagtagaaaagtaggtcattcttggatctacttttttttagcctta 3901 ttaatatttttccctattagaaaccacaattactccctctattaacccttcacttactag 3961 accagaaaagaacttattccagataagctttgaatatcaattcttacataaactttaggc 4021 aaacagggaatagtctagtcaccaaaggaccattctcttgccaatgctgcattccttttg 4081 cacttttggattccatatttatcccaaatgctgttgggcacccctagaaataccttgatg 4141 ttttttctatttatatgcctgcctttggtacttaattttacaaatgctgtaatataaagc 4201 atatcaagtttatgtgatacgtatcattgcaagagaatttgtttcaagatttttttttaa 4261 tgttccagaagatggccaatagagaacattcaagggaaatggggaaacataatttagaga 4321 acaagaacaaaccatgtctcaaatttttttaaaaaaaattaatggttttaaatatatgct 4381 atagggacgttccatgcccaggttaacaaagaactgtgatatatagagtgtctaattaca 4441 aaatcatatacgatttatttaattctcttctgtattgtaacttagatgattcccaaggac 4501 tctaataaaaaatcacttcattgtatttggaaacaaaaacatcattcattaattacttat 4561 tttctttccataggttttaatattttgagagtgtcttttttatttcattcatgaactttt 4621 gtatttttcatttttcatttgatttgtaaatttacttatgttaaaaataaaccatttatt 4681 ttcagctttgaattttaaaaaaaaaaaaaaaaaa HumanIL1RAPmRNAVariant5 (SEQIDNO:118) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaaggatgacacttc 181 tgtggtgtgtagtgagtctctacttttatggaatcctgcaaagtgatgcctcagaacgct 241 gcgatgactggggactagacaccatgaggcaaatccaagtgtttgaagatgagccagctc 301 gcatcaagtgcccactctttgaacacttcttgaaattcaactacagcacagcccattcag 361 ctggccttactctgatctggtattggactaggcaggaccgggaccttgaggagccaatta 421 acttccgcctccccgagaaccgcattagtaaggagaaagatgtgctgtggttccggccca 481 ctctcctcaatgacactggcaactatacctgcatgttaaggaacactacatattgcagca 541 aagttgcatttcccttggaagttgttcaaaaagacagctgtttcaattcccccatgaaac 601 tcccagtgcataaactgtatatagaatatggcattcagaggatcacttgtccaaatgtag 661 atggatattttccttccagtgtcaaaccgactatcacttggtatatgggctgttataaaa 721 tacagaattttaataatgtaatacccgaaggtatgaacttgagtttcctcattgccttaa 781 tttcaaataatggaaattacacatgtgttgttacatatccagaaaatggacgtacgtttc 841 atctcaccaggactctgactgtaaaggtagtaggctctccaaaaaatgcagtgccccctg 901 tgatccattcacctaatgatcatgtggtctatgagaaagaaccaggagaggagctactca 961 ttccctgtacggtctattttagttttctgatggattctcgcaatgaggtttggtggacca 1021 ttgatggaaaaaaacctgatgacatcactattgatgtcaccattaacgaaagtataagtc 1081 atagtagaacagaagatgaaacaagaactcagattttgagcatcaagaaagttacctctg 1141 aggatctcaagcgcagctatgtctgtcatgctagaagtgccaaaggcgaagttgccaaag 1201 cagccaaggtgaagcagaaaggtaatagatgcggtcagtgatgaatctctcagctccaaa 1261 ttaacattgtggtgaataaggacaaaaggagagattgagaacaagagagctccagcacct 1321 agcccgacggcatctaacccatagtaatgaatcaaacttaaatgaaaaatatgaaagttt 1381 tcatctatgtaagatactcaaaatattgtttctgatattgttagtaccgtaatgcccaaa 1441 tgtagctaaaaaaatcgacgtgagtacagtgagacacaattttgtgtctgtacaattatg 1501 aaaaattaaaaacaaagaaaatattcaaagctaccaaagatagaaaaaactggtagagcc 1561 acatattgttggtgaattattaagacccttttaaaaatcattcatggtagacttcaagag 1621 tcataaaaaagattgcatcatctgacctaagactttcggaatttttcctgaacaaataac 1681 agaaagggaattatataccttttaatattattagaagcattatctgtagttgtaaaacat 1741 tattaatagcagccatccaattgtatgcaactaattaaggtattgaatgtttattttcca 1801 aaaatgcataattataatattattttaaacactatgtatcaatatttaagcaggtttata 1861 atataccagcagccacaattgctaaaatgaaaatcatttaaattatgattttaaatggta 1921 taaacatgatttctatgttgatagtactatattattctacaataaatggaaattataaag 1981 ccttcttgtcagaagtgctgctcctaaaaaaaaaaaaaaaaaaaaaa HumanIL1RAPmRNAVariant6 (SEQIDNO:119) 1 aaagggggaaaagaaagtgcggcggaaagtaagaggctcactggggaagactgccgggat 61 ccaggtctccggggtccgctttggccagaggcgcggaaggaagcagtgcccggcgacact 121 gcacccatcccggctgcttttgctgcgccctctcagcttcccaagaaaggcatcgtcatg 181 tgatcatcacctaagaactagaacatcagcaggccctagaagcctcactcttgcccctcc 241 ctttaatatctcaaaggatgacacttctgtggtgtgtagtgagtctctacttttatggaa 301 tcctgcaaagtgatgcctcagaacgctgcgatgactggggactagacaccatgaggcaaa 361 tccaagtgtttgaagatgagccagctcgcatcaagtgcccactctttgaacacttcttga 421 aattcaactacagcacagcccattcagctggccttactctgatctggtattggactaggc 481 aggaccgggaccttgaggagccaattaacttccgcctccccgagaaccgcattagtaagg 541 agaaagatgtgctgtggttccggcccactctcctcaatgacactggcaactatacctgca 601 tgttaaggaacactacatattgcagcaaagttgcatttcccttggaagttgttcaaaaag 661 acagctgtttcaattcccccatgaaactcccagtgcataaactgtatatagaatatggca 721 ttcagaggatcacttgtccaaatgtagatggatattttccttccagtgtcaaaccgacta 781 tcacttggtatatgggctgttataaaatacagaattttaataatgtaatacccgaaggta 841 tgaacttgagtttcctcattgccttaatttcaaataatggaaattacacatgtgttgtta 901 catatccagaaaatggacgtacgtttcatctcaccaggactctgactgtaaaggtagtag 961 gctctccaaaaaatgcagtgccccctgtgatccattcacctaatgatcatgtggtctatg 1021 agaaagaaccaggagaggagctactcattccctgtacggtctattttagttttctgatgg 1081 attctcgcaatgaggtttggtggaccattgatggaaaaaaacctgatgacatcactattg 1141 atgtcaccattaacgaaagtataagtcatagtagaacagaagatgaaacaagaactcaga 1201 ttttgagcatcaagaaagttacctctgaggatctcaagcgcagctatgtctgtcatgcta 1261 gaagtgccaaaggcgaagttgccaaagcagccaaggtgaagcagaaagtgccagctccaa 1321 gatacacagtggaactggcttgtggttttggagccacagtcctgctagtggtgattctca 1381 ttgttgtttaccatgtttactggctagagatggtcctattttaccgggctcattttggaa 1441 cagatgaaaccattttagatggaaaagagtatgatatttatgtatcctatgcaaggaatg 1501 cggaagaagaagaatttgtattactgaccctccgtggagttttggagaatgaatttggat 1561 acaagctgtgcatctttgaccgagacagtctgcctgggggaaatacagtggaagcagttt 1621 ttgatttcattcagagaagcagaaggatgattgttgttctgagccctgactatgtgacag 1681 aaaagagcatcagcatgctggagtttaaactgggtgtcatgtgccagaactccattgcca 1741 ccaagctcattgtggttgagtaccgtccccttgagcacccgcacccaggcattcttcagc 1801 tcaaagagtctgtgtcttttgtgagctggaagggagaaaagtccaaacattctggctcta 1861 aattctggaaagctttgcggttggctcttcccctgagaagtctgagtgccagttctggct 1921 ggaatgagagctgctcttcccagtctgacatcagtctggatcacgttcaaaggaggagaa 1981 gtcgtttgaaagagcccccagaacttcagagctcagagagggctgcaggtagccctccag 2041 ccccaggcacaatgtccaagcaccgagggaagtcctccgccacctgccgctgttgtgtca 2101 cctactgtgaaggagagaatcaccttaggaacaagagccgggcagagattcataaccagc 2161 cccagtgggagacacacctctgtaagcctgttccccaagagtcagaaactcaatggatac 2221 aaaatggcaccagattggaaccccctgctccccagatctcagcccttgctcttcatcatt 2281 tcacggacttatccaataacaacgacttttatatcctataattactgtgtgtggtgggtg 2341 gtggctactatctctaccaaccctctgtatgtcatgaacctgtgggaaaatctgacattt 2401 ttatcatctaatggactatcagatttctgtcccctttattgatttttaaaaactatttat 2461 ttctaggagacaaaagacctgaaggacctgaatccagaattattgcctctaaaggcctca 2521 gaagagcacactcttcttgggccctagaaggtcagtatgtgaaagttgcctaaagtctga 2581 tcctctatcttgtccaatggtttaaaactgagctaagaatttaaatgtgtttcttttcag 2641 tgagttgatcaacctcacattataagtcagtcaggtgtacttgggctatgatgcttacag 2701 ggtgtatgcattcccagggagcagcatggaaaggagctggttctggtggaagctgtagga 2761 cgaagctcaacagaaaacctacagcacatttttcctcaaagaaccaaacatacccaccca 2821 gggatacatggcgttctctgtctcactgtaaactagtgttctctaaactgcctaacattg 2881 ttagcatcaataaaattctatttttacgtcaaaaaaaaaaaaaaaaaaaa HumanIL-18RmRNAVariant1 (SEQIDNO:120) 1 tcaggaggcggagatcgctgcttctcacctactttctgaacttggcctccgcagtcgcga 61 cctggcgtgaaggaggagctgccgcccccgccccagcctcggggacgcctctctgaagag 121 aagccatttgaagcagaatccaaaccatgaattgtagagaattacccttgaccctttggg 181 tgcttatatctgtaagcactgcagaatcttgtacttcacgtccccacattactgtggttg 241 aaggggaacctttctatctgaaacattgctcgtgttcacttgcacatgagattgaaacaa 301 ccaccaaaagctggtacaaaagcagtggatcacaggaacatgtggagctgaacccaagga 361 gttcctcgagaattgctttgcatgattgtgttttggagttttggccagttgagttgaatg 421 acacaggatcttactttttccaaatgaaaaattatactcagaaatggaaattaaatgtca 481 tcagaagaaataaacacagctgtttcactgaaagacaagtaactagtaaaattgtggaag 541 ttaaaaaattttttcagataacctgtgaaaacagttactatcaaacactggtcaacagca 601 catcattgtataagaactgtaaaaagctactactggagaacaataaaaacccaacgataa 661 agaagaacgccgagtttgaagatcaggggtattactcctgcgtgcatttccttcatcata 721 atggaaaactatttaatatcaccaaaaccttcaatataacaatagtggaagatcgcagta 781 atatagttccggttcttcttggaccaaagcttaaccatgttgcagtggaattaggaaaaa 841 acgtaaggctcaactgctctgctttgctgaatgaagaggatgtaatttattggatgttcg 901 gggaagaaaatggatcggatcctaatatacatgaagagaaagaaatgagaattatgactc 961 cagaaggcaaatggcatgcttcaaaagtattgagaattgaaaatattggtgaaagcaatc 1021 taaatgttttatataattgcactgtggccagcacgggaggcacagacaccaaaagcttca 1081 tcttggtgagaaaagcagacatggctgatatcccaggccacgtcttcacaagaggaatga 1141 tcatagctgttttgatcttggtggcagtagtgtgcctagtgactgtgtgtgtcatttata 1201 gagttgacttggttctattttatagacatttaacgagaagagatgaaacattaacagatg 1261 gaaaaacatatgatgcttttgtgtcttacctaaaagaatgccgacctgaaaatggagagg 1321 agcacacctttgctgtggagattttgcccagggtgttggagaaacattttgggtataagt 1381 tatgcatatttgaaagggatgtagtgcctggaggagctgttgttgatgaaatccactcac 1441 tgatagagaaaagccgaagactaatcattgtcctaagtaaaagttatatgtctaatgagg 1501 tcaggtatgaacttgaaagtggactccatgaagcattggtggaaagaaaaattaaaataa 1561 tcttaattgaatttacacctgttactgacttcacattcttgccccaatcactaaagcttt 1621 tgaaatctcacagagttctgaagtggaaggccgataaatctctttcttataactcaaggt 1681 tctggaagaaccttctttacttaatgcctgcaaaaacagtcaagccaggtagagacgaac 1741 cggaagtcttgcctgttctttccgagtcttaatcttcagaaacagtgaacgccaaaaaga 1801 actcaagatattctggggactgagcatatgaacctgttcataacaaaggctgtgactcga 1861 aataattaactttgtcaaaatcctgctcacaatttgaagatgaaacttgtcattaggttg 1921 gcgggaatgagactaaagattgcgctgtgggctgtggtcacgtgctcccagaagacctgg 1981 aattcaaaagaaatggagctattctttttctccctctttcataactggatgcagctgctc 2041 atactcaatcccatattcagcaagtgtgaagctggacgtgatgcaaaataaccgatgccc 2101 tacaaaaagggcgcatctttaagagttttaatgccagtgcttaattcgaatgaggggatt 2161 ttaagtgtctgaagaggcattttctagggaccagtgggtgactgagtaactgaaatgctg 2221 ctttcactccctaacaccatggatctggttgtgcataggatgtgggaggaggggctggca 2281 gggccgccttcagaggctgcagggcctcagcctcaggatgcatttaatgtatcctggcca 2341 cagttgcagccaacggttcttgaaagctcggtaaggccctgcaacgcagagcctgcttat 2401 gtggatctatttatgggaacttcttaaaaggaccccagaatagctctttatctttcacaa 2461 gagacacaaattctaattgagttaattatctgggcctttcactttggatgctctgaaaca 2521 tttgttgattttgtgtgaatgtttatatcaaaatgtttgccaggttgtattagccattga 2581 atagcaaaaaactgatagttacttgcttgttttttaaaaattacatattaaaaatgccct 2641 tggcataaggcagcatggtgtggcagttaagagatgggctgtgcagcccatcctgagctc 2701 cagtcctgagtttgctacttacttctgtggcctctggaaccttatccaacctcttggtgc 2761 ttcagtttcctcatctgtgaaattagaatttataataattgcacctacctcccaggggta 2821 actaaatgaataaatataataaagtacttacagtggttcctgacacagactcagcactcc 2881 gtcagtgttgccatgactatttttattatcattattaatgattacttagatcaattattt 2941 agcagtggactaatggaagctacagagcagggaagggaagcagatctagggaggaaggca 3001 gttttgatttgaggaggtttgcacatgtagagaagcatactggagaagcatatccagagg 3061 gcgaaagatatctctccattgtgcatctgcctcttttgacgttggaagacacatgtctta 3121 ctccccaaagggagcccagcactgggagccttcttgatgatctcaaaaataatagctatt 3181 caagaaaatcaccaagtgactgtgaaaccgtcagttcggaaggctggttagaacatgtgg 3241 gagcaacatgaatgttctacaaaagtttaaagcagagattgtttcaaatgggtgtagtag 3301 atattactgaaaaccaaaaaagagtgagattgtcagtgtaagaatgtgatttaatgtttg 3361 tagtgcttacaattttgtgtaccaactggatgactaaaaagagtaaaataatttaattaa 3421 tagctcatattttatgtgtgaaaacatgttagtgaacatatataatcaaaatagatttca 3481 ttgctattgcatagtctctaatacatagaatgattttgcttttctcttttattatacttg 3541 ctttaaaatacttgaaatatattttgcattaaatgcatttcaagttaaatgtcttaaatg 3601 tatacattagatgtgtgttttaaaatgcataaaacacgttgaaatacattaatgaaccat 3661 t HumanIL-18RmRNAVariant2 (SEQIDNO:121) 1 tcaggaggcggagatcgctgcttctcacctactttctgaacttggcctccgcagtcgcga 61 cctggcgtgaaggaggagctgccgcccccgccccagcctcggggacgcctctctgaagag 121 aagccatttgaagcagaatccaaaccatgaattgtagagaattacccttgaccctttggg 181 tgcttatatctgtaagcactgcagaaattatactcagaaatggaaattaaatgtcatcag 241 aagaaataaacacagctgtttcactgaaagacaagtaactagtaaaattgtggaagttaa 301 aaaattttttcagataacctgtgaaaacagttactatcaaacactggtcaacagcacatc 361 attgtataagataggaccacctatttgcaggaaaacaagctcagggctccactgattcta 421 cattatgaactgtaaaaagctactactggagaacaataaaaacccaacgataaagaagaa 481 cgccgagtttgaagatcaggggtattactcctgcgtgcatttccttcatcataatggaaa 541 actatttaatatcaccaaaaccttcaatataacaatagtggaagatcgcagtaatatagt 601 tccggttcttcttggaccaaagcttaaccatgttgcagtggaattaggaaaaaacgtaag 661 gctcaactgctctgctttgctgaatgaagaggatgtaatttattggatgttcggggaaga 721 aaatggatcggatcctaatatacatgaagagaaagaaatgagaattatgactccagaagg 781 caaatggcatgcttcaaaagtattgagaattgaaaatattggtgaaagcaatctaaatgt 841 tttatataattgcactgtggccagcacgggaggcacagacaccaaaagcttcatcttggt 901 gagaaaagacatggctgatatcccaggccacgtcttcacaagaggaatgatcatagctgt 961 tttgatcttggtggcagtagtgtgcctagtgactgtgtgtgtcatttatagagttgactt 1021 ggttctattttatagacatttaacgagaagagatgaaacattaacagatggaaaaacata 1081 tgatgcttttgtgtcttacctaaaagaatgccgacctgaaaatggagaggagcacacctt 1141 tgctgtggagattttgcccagggtgttggagaaacattttgggtataagttatgcatatt 1201 tgaaagggatgtagtgcctggaggagctgttgttgatgaaatccactcactgatagagaa 1261 aagccgaagactaatcattgtcctaagtaaaagttatatgtctaatgaggtcaggtatga 1321 acttgaaagtggactccatgaagcattggtggaaagaaaaattaaaataatcttaattga 1381 atttacacctgttactgacttcacattcttgccccaatcactaaagcttttgaaatctca 1441 cagagttctgaagtggaaggccgataaatctctttcttataactcaaggttctggaagaa 1501 ccttctttacttaatgcctgcaaaaacagtcaagccaggtagagacgaaccggaagtctt 1561 gcctgttctttccgagtcttaatcttcagaaacagtgaacgccaaaaagaactcaagata 1621 ttctggggactgagcatatgaacctgttcataacaaaggctgtgactcgaaataattaac 1681 tttgtcaaaatcctgctcacaatttgaagatgaaacttgtcattaggttggcgggaatga 1741 gactaaagattgcgctgtgggctgtggtcacgtgctcccagaagacctggaattcaaaag 1801 aaatggagctattctttttctccctctttcataactggatgcagctgctcatactcaatc 1861 ccatattcagcaagtgtgaagctggacgtgatgcaaaataaccgatgccctacaaaaagg 1921 gcgcatctttaagagttttaatgccagtgcttaattcgaatgaggggattttaagtgtct 1981 gaagaggcattttctagggaccagtgggtgactgagtaactgaaatgctgctttcactcc 2041 ctaacaccatggatctggttgtgcataggatgtgggaggaggggctggcagggccgcctt 2101 cagaggctgcagggcctcagcctcaggatgcatttaatgtatcctggccacagttgcagc 2161 caacggttcttgaaagctcggtaaggccctgcaacgcagagcctgcttatgtggatctat 2221 ttatgggaacttcttaaaaggaccccagaatagctctttatctttcacaagagacacaaa 2281 ttctaattgagttaattatctgggcctttcactttggatgctctgaaacatttgttgatt 2341 ttgtgtgaatgtttatatcaaaatgtttgccaggttgtattagccattgaatagcaaaaa 2401 actgatagttacttgcttgttttttaaaaattacatattaaaaatgcccttggcataagg 2461 cagcatggtgtggcagttaagagatgggctgtgcagcccatcctgagctccagtcctgag 2521 tttgctacttacttctgtggcctctggaaccttatccaacctcttggtgcttcagtttcc 2581 tcatctgtgaaattagaatttataataattgcacctacctcccaggggtaactaaatgaa 2641 taaatataataaagtacttacagtggttcctgacacagactcagcactccgtcagtgttg 2701 ccatgactatttttattatcattattaatgattacttagatcaattatttagcagtggac 2761 taatggaagctacagagcagggaagggaagcagatctagggaggaaggcagttttgattt 2821 gaggaggtttgcacatgtagagaagcatactggagaagcatatccagagggcgaaagata 2881 tctctccattgtgcatctgcctcttttgacgttggaagacacatgtcttactccccaaag 2941 ggagcccagcactgggagccttcttgatgatctcaaaaataatagctattcaagaaaatc 3001 accaagtgactgtgaaaccgtcagttcggaaggctggttagaacatgtgggagcaacatg 3061 aatgttctacaaaagtttaaagcagagattgtttcaaatgggtgtagtagatattactga 3121 aaaccaaaaaagagtgagattgtcagtgtaagaatgtgatttaatgtttgtagtgcttac 3181 aattttgtgtaccaactggatgactaaaaagagtaaaataatttaattaatagctcatat 3241 tttatgtgtgaaaacatgttagtgaacatatataatcaaaatagatttcattgctattgc 3301 atagtctctaatacatagaatgattttgcttttctcttttattatacttgctttaaaata 3361 cttgaaatatattttgcattaaatgcatttcaagttaaatgtcttaaatgtatacattag 3421 atgtgtgttttaaaatgcataaaacacgttgaaatacattaatgaaccatt HumanIL-1RL2mRNA (SEQIDNO:122) 1 cccgcccacggtggcggggaaatacctaggcatggaagtggcatgacagggctcgtgtcc 61 ctgtcatattttccactctccacgaggtcctgcgcgcttcaatcctgcaggcagcccggt 121 ttggggatgtggtccttgctgctctgcgggttgtccatcgcccttccactgtctgtcaca 181 gcagatggatgcaaggacatttttatgaaaaatgagatactttcagcaagccagcctttt 241 gcttttaattgtacattccctcccataacatctggggaagtcagtgtaacatggtataaa 301 aattctagcaaaatcccagtgtccaaaatcatacagtctagaattcaccaggacgagact 361 tggattttgtttctccccatggaatggggggactcaggagtctaccaatgtgttataaag 421 ggtagagacagctgtcatagaatacatgtaaacctaactgtttttgaaaaacattggtgt 481 gacacttccataggtggtttaccaaatttatcagatgagtacaagcaaatattacatctt 541 ggaaaagatgatagtctcacatgtcatctgcacttcccgaagagttgtgttttgggtcca 601 ataaagtggtataaggactgtaacgagattaaaggggagcggttcactgttttggaaacc 661 aggcttttggtgagcaatgtctcggcagaggacagagggaactacgcgtgtcaagccata 721 ctgacacactcagggaagcagtacgaggttttaaatggcatcactgtgagcattacagaa 781 agagctggatatggaggaagtgtccctaaaatcatttatccaaaaaatcattcaattgaa 841 gtacagcttggtaccactctgattgtggactgcaatgtaacagacaccaaggataataca 901 aatctacgatgctggagagtcaataacactttggtggatgattactatgatgaatccaaa 961 cgaatcagagaaggggtggaaacccatgtctcttttcgggaacataatttgtacacagta 1021 aacatcaccttcttggaagtgaaaatggaagattatggccttcctttcatgtgccacgct 1081 ggagtgtccacagcatacattatattacagctcccagctccggattttcgagcttacttg 1141 ataggagggcttatcgccttggtggctgtggctgtgtctgttgtgtacatatacaacatt 1201 tttaagatcgacattgttctttggtatcgaagtgccttccattctacagagaccatagta 1261 gatgggaagctgtatgacgcctatgtcttataccccaagccccacaaggaaagccagagg 1321 catgccgtggatgccctggtgttgaatatcctgcccgaggtgttggagagacaatgtgga 1381 tataagttgtttatattcggcagagatgaattccctggacaagccgtggccaatgtcatc 1441 gatgaaaacgttaagctgtgcaggaggctgattgtcattgtggtccccgaatcgctgggc 1501 tttggcctgttgaagaacctgtcagaagaacaaatcgcggtctacagtgccctgatccag 1561 gacgggatgaaggttattctcattgagctggagaaaatcgaggactacacagtcatgcca 1621 gagtcaattcagtacatcaaacagaagcatggtgccatccggtggcatggggacttcacg 1681 gagcagtcacagtgtatgaagaccaagttttggaagacagtgagataccacatgccgccc 1741 agaaggtgtcggccgtttcctccggtccagctgctgcagcacacaccttgctaccgcacc 1801 gcaggcccagaactaggctcaagaagaaagaagtgtactctcacgactggctaagacttg 1861 ctggactgacacctatggctggaagatgacttgttttgctccatgtctcctcattcctac 1921 acctattttctgctgcaggatgaggctagggttagcattctaga HumanIL1RL1mRNAVariant1 (SEQIDNO:123) 1 aaagagaggctggctgttgtatttagtaaagctataaagctgtaagagaaattggctttc 61 tgagttgtgaaactgtgggcagaaagttgaggaagaaagaactcaagtacaacccaatga 121 ggttgagatataggctactcttcccaactcagtcttgaagagtatcaccaactgcctcat 181 gtgtggtgaccttcactgtcgtatgccagtgactcatctggagtaatctcaacaacgagt 241 taccaatacttgctcttgattgataaacagaatggggttttggatcttagcaattctcac 301 aattctcatgtattccacagcagcaaagtttagtaaacaatcatggggcctggaaaatga 361 ggctttaattgtaagatgtcctagacaaggaaaacctagttacaccgtggattggtatta 421 ctcacaaacaaacaaaagtattcccactcaggaaagaaatcgtgtgtttgcctcaggcca 481 acttctgaagtttctaccagctgcagttgctgattctggtatttatacctgtattgtcag 541 aagtcccacattcaataggactggatatgcgaatgtcaccatatataaaaaacaatcaga 601 ttgcaatgttccagattatttgatgtattcaacagtatctggatcagaaaaaaattccaa 661 aatttattgtcctaccattgacctctacaactggacagcacctcttgagtggtttaagaa 721 ttgtcaggctcttcaaggatcaaggtacagggcgcacaagtcatttttggtcattgataa 781 tgtgatgactgaggacgcaggtgattacacctgtaaatttatacacaatgaaaatggagc 841 caattatagtgtgacggcgaccaggtccttcacggtcaaggatgagcaaggcttttctct 901 gtttccagtaatcggagcccctgcacaaaatgaaataaaggaagtggaaattggaaaaaa 961 cgcaaacctaacttgctctgcttgttttggaaaaggcactcagttcttggctgccgtcct 1021 gtggcagcttaatggaacaaaaattacagactttggtgaaccaagaattcaacaagagga 1081 agggcaaaatcaaagtttcagcaatgggctggcttgtctagacatggttttaagaatagc 1141 tgacgtgaaggaagaggatttattgctgcagtacgactgtctggccctgaatttgcatgg 1201 cttgagaaggcacaccgtaagactaagtaggaaaaatccaattgatcatcatagcatcta 1261 ctgcataattgcagtatgtagtgtatttttaatgctaatcaatgtcctggttatcatcct 1321 aaaaatgttctggattgaggccactctgctctggagagacatagctaaaccttacaagac 1381 taggaatgatggaaagctctatgatgcttatgttgtctacccacggaactacaaatccag 1441 tacagatggggccagtcgtgtagagcactttgttcaccagattctgcctgatgttcttga 1501 aaataaatgtggctataccttatgcatttatgggagagatatgctacctggagaagatgt 1561 agtcactgcagtggaaaccaacatacgaaagagcaggcggcacattttcatcctgacccc 1621 tcagatcactcacaataaggagtttgcctacgagcaggaggttgccctgcactgtgccct 1681 catccagaacgacgccaaggtgatacttattgagatggaggctctgagcgagctggacat 1741 gctgcaggctgaggcgcttcaggactccctccagcatcttatgaaagtacaggggaccat 1801 caagtggagggaggaccacattgccaataaaaggtccctgaattctaaattctggaagca 1861 cgtgaggtaccaaatgcctgtgccaagcaaaattcccagaaaggcctctagtttgactcc 1921 cttggctgcccagaagcaatagtgcctgctgtgatgtgcaaaggcatctgagtttgaagc 1981 tttcctgacttctcctagctggcttatgcccctgcactgaagtgtgaggagcaggaatat 2041 taaagggattcaggcctc HumanIL1RL1mRNAVariant2 (SEQIDNO:124) 1 agtctatgaggagggacctacaaagactggaaactattcttagctccgtcactgactcca 61 agttcatcccctctgtctttcagtttggttgagatataggctactcttcccaactcagtc 121 ttgaagagtatcaccaactgcctcatgtgtggtgaccttcactgtcgtatgccagtgact 181 catctggagtaatctcaacaacgagttaccaatacttgctcttgattgataaacagaatg 241 gggttttggatcttagcaattctcacaattctcatgtattccacagcagcaaagtttagt 301 aaacaatcatggggcctggaaaatgaggctttaattgtaagatgtcctagacaaggaaaa 361 cctagttacaccgtggattggtattactcacaaacaaacaaaagtattcccactcaggaa 421 agaaatcgtgtgtttgcctcaggccaacttctgaagtttctaccagctgcagttgctgat 481 tctggtatttatacctgtattgtcagaagtcccacattcaataggactggatatgcgaat 541 gtcaccatatataaaaaacaatcagattgcaatgttccagattatttgatgtattcaaca 601 gtatctggatcagaaaaaaattccaaaatttattgtcctaccattgacctctacaactgg 661 acagcacctcttgagtggtttaagaattgtcaggctcttcaaggatcaaggtacagggcg 721 cacaagtcatttttggtcattgataatgtgatgactgaggacgcaggtgattacacctgt 781 aaatttatacacaatgaaaatggagccaattatagtgtgacggcgaccaggtccttcacg 841 gtcaaggatgagcaaggcttttctctgtttccagtaatcggagcccctgcacaaaatgaa 901 ataaaggaagtggaaattggaaaaaacgcaaacctaacttgctctgcttgttttggaaaa 961 ggcactcagttcttggctgccgtcctgtggcagcttaatggaacaaaaattacagacttt 1021 ggtgaaccaagaattcaacaagaggaagggcaaaatcaaagtttcagcaatgggctggct 1081 tgtctagacatggttttaagaatagctgacgtgaaggaagaggatttattgctgcagtac 1141 gactgtctggccctgaatttgcatggcttgagaaggcacaccgtaagactaagtaggaaa 1201 aatccaagtaaggagtgtttctgagactttgatcacctgaactttctctagcaagtgtaa 1261 gcagaatggagtgtggttccaagagatccatcaagacaatgggaatggcctgtgccataa 1321 aatgtgcttctcttcttcgggatgttgtttgctgtctgatctttgtagactgttcctgtt 1381 tgctgggagcttctctgctgcttaaattgttcgtcctcccccactccctcctatcgttgg 1441 tttgtctagaacactcagctgcttctttggtcatccttgttttctaactttatgaactcc 1501 ctctgtgtcactgtatgtgaaaggaaatgcaccaacaaccgtaaactgaacgtgttcttt 1561 tgtgctcttttataacttgcattacatgttgtaagcatggtccgttctatacctttttct 1621 ggtcataatgaacactcattttgttagcgagggtggtaaagtgaacaaaaaggggaagta 1681 tcaaactactgccatttcagtgagaaaatcctaggtgctactttataataagacatttgt 1741 taggccattcttgcattgatataaagaaatacctgagactgggtgatttatatgaaaaga 1801 ggtttaattggctcacagttctgcaggctgtatgggaagcatggcggcatctgcttctgg 1861 ggacacctcaggagctttactcatggcagaaggcaaagcaaaggcaggcacttcacacag 1921 taaaagcaggagcgagagagaggtgccacactgaaacagccagatctcatgagaagtcac 1981 tcactattgcaaggacagcatcaaagagatggtgctaaaccattcatgatgaactcaccc 2041 ccatgatccaatcacctcccaccaggctccacctcgaatactggggattaccattcagca 2101 tgagatttgggcaggaacacagacccaaaccataccacacacattatcattgttaaactt 2161 tgtaaagtatttaaggtacatggaacacacgggaagtctggtagctcagcccatttcttt 2221 attgcatctgttattcaccatgtaattcaggtaccacgtattccagggagcctttcttgg 2281 ccctcagtttgcagtatacacactttccaagtactcttgtagcatcctgtttgtatcata 2341 gcactggtcacattgccttacctaaatctgtttgacagtctgctcaacacgactgcaagc 2401 tccatgagggcagggacatcatctcttccatctttgggtccttagtgcaatacctggcag 2461 ctagccagtgctcagctaaatatttgttgactgaataaatgaatgcacaaccaaattatt 2521 gataccaaatgttttttttgtgtacatttctacttctctagctataagtcttaattatac 2581 aacaaaatactatttttatatttatgtttggtaaattcaataactttcctcatcatttgg 2641 aaagtcaaattgtttattgcttccctacagttttttctgaatctagcaggattttaatga 2701 tatcattataatttgacacaataaaaggacaacatgaaactgatgaatctttattgggtt 2761 aatttcagacactatataatcttttaaaaatgtaacattcttttttatatataaataatt 2821 ggtggcatcacaaatagccaaagcagggtggagagagtgatccttcctgggtgcaggcaa 2881 gaaggggatatgttttctacagagttttcaaaacagtgataaagctgtctacaagtcatt 2941 gtgctttttatcatcactatgcccagacaatgtgaaacatcagagatgaagtgctcttcc 3001 cacagaggtggactgatccttctccccactcccttggtgtgtctctgaatgcaatgttgt 3061 cttggaaaacagctttccaagcatttcactcctgagcacttgccagtttcctcacttgtt 3121 cttcacatatccaggcaaagacatcctgtttgctatatgaagcattgtatcccgtataaa 3181 aggaaggaaagagagaaatatatttttacactcatcactcctcaggggctgtacaatcat 3241 gtagaaattgtttaatgtgcctgtcaaatagccaaagagtgttaaaccctgagttcccac 3301 ccatgtgtgtggtatggttaggattcatccagatacacagagagaggcacaacaggagga 3361 gaaaggataggggtgtggggacagcgggcccccaatatggtgtaatcgtggcaggtctct 3421 gcctgaagtgctatgtggggtttttcttgttttaattttgactttaacccctgatttgta 3481 agtttttcataaaataaacagaatcataactcatgtagatggctataagtgccgtagtgt 3541 tctgtgggtctctggtgtctgccagtgataagtgtggcaccccaggaaggctgtggaccc 3601 catcaaggtgctatgtgagggccatgcttggggtggtggtgggcccagtagaccctgcag 3661 ccatccatccagcctgcccactcacactgcccttgtgtactcctgctttgctacgttatc 3721 attgatcaatgtccctggttacctatgtgtttgaattatcttcgtgttacaggtgtttaa 3781 tgattttgctccttctagcttatttgtatttcacctgtttttctttaaatcaacatggtt 3841 acactctgtttcagcaactgtataaattaaacacaaattattactactgctaaaaaaaaa 3901 aaaaaaaaa HumanIL1RL1mRNAVariant3 (SEQIDNO:125) 1 aaagagaggctggctgttgtatttagtaaagctataaagctgtaagagaaattggctttc 61 tgagttgtgaaactgtgggcagaaagttgaggaagaaagaactcaagtacaacccaatga 121 gggccaacttctgaagtttctaccagctgcagttgctgattctggtatttatacctgtat 181 tgtcagaagtcccacattcaataggactggatatgcgaatgtcaccatatataaaaaaca 241 atcagattgcaatgttccagattatttgatgtattcaacagtatctggatcagaaaaaaa 301 ttccaaaatttattgtcctaccattgacctctacaactggacagcacctcttgagtggtt 361 taagaattgtcaggctcttcaaggatcaaggtacagggcgcacaagtcatttttggtcat 421 tgataatgtgatgactgaggacgcaggtgattacacctgtaaatttatacacaatgaaaa 481 tggagccaattatagtgtgacggcgaccaggtccttcacggtcaaggatgagcaaggctt 541 ttctctgtttccagtaatcggagcccctgcacaaaatgaaataaaggaagtggaaattgg 601 aaaaaacgcaaacctaacttgctctgcttgttttggaaaaggcactcagttcttggctgc 661 cgtcctgtggcagcttaatggaacaaaaattacagactttggtgaaccaagaattcaaca 721 agaggaagggcaaaatcaaagtttcagcaatgggctggcttgtctagacatggttttaag 781 aatagctgacgtgaaggaagaggatttattgctgcagtacgactgtctggccctgaattt 841 gcatggcttgagaaggcacaccgtaagactaagtaggaaaaatccaagtaaggagtgttt 901 ctgagactttgatcacctgaactttctctagcaagtgtaagcagaatggagtgtggttcc 961 aagagatccatcaagacaatgggaatggcctgtgccataaaatgtgcttctcttcttcgg 1021 gatgttgtttgctgtctgatctttgtagactgttcctgtttgctgggagcttctctgctg 1081 cttaaattgttcgtcctcccccactccctcctatcgttggtttgtctagaacactcagct 1141 gcttctttggtcatccttgttttctaactttatgaactccctctgtgtcactgtatgtga 1201 aaggaaatgcaccaacaaccgtaaactgaacgtgttcttttgtgctcttttataacttgc 1261 attacatgttgtaagcatggtccgttctatacctttttctggtcataatgaacactcatt 1321 ttgttagcgagggtggtaaagtgaacaaaaaggggaagtatcaaactactgccatttcag 1381 tgagaaaatcctaggtgctactttataataagacatttgttaggccattcttgcattgat 1441 ataaagaaatacctgagactgggtgatttatatgaaaagaggtttaattggctcacagtt 1501 ctgcaggctgtatgggaagcatggcggcatctgcttctggggacacctcaggagctttac 1561 tcatggcagaaggcaaagcaaaggcaggcacttcacacagtaaaagcaggagcgagagag 1621 aggtgccacactgaaacagccagatctcatgagaagtcactcactattgcaaggacagca 1681 tcaaagagatggtgctaaaccattcatgatgaactcacccccatgatccaatcacctccc 1741 accaggctccacctcgaatactggggattaccattcagcatgagatttgggcaggaacac 1801 agacccaaaccataccacacacattatcattgttaaactttgtaaagtatttaaggtaca 1861 tggaacacacgggaagtctggtagctcagcccatttctttattgcatctgttattcacca 1921 tgtaattcaggtaccacgtattccagggagcctttcttggccctcagtttgcagtataca 1981 cactttccaagtactcttgtagcatcctgtttgtatcatagcactggtcacattgcctta 2041 cctaaatctgtttgacagtctgctcaacacgactgcaagctccatgagggcagggacatc 2101 atctcttccatctttgggtccttagtgcaatacctggcagctagccagtgctcagctaaa 2161 tatttgttgactgaataaatgaatgcacaaccaaattattgataccaaatgttttttttg 2221 tgtacatttctacttctctagctataagtcttaattatacaacaaaatactatttttata 2281 tttatgtttggtaaattcaataactttcctcatcatttggaaagtcaaattgtttattgc 2341 ttccctacagttttttctgaatctagcaggattttaatgatatcattataatttgacaca 2401 ataaaaggacaacatgaaactgatgaatctttattgggttaatttcagacactatataat 2461 cttttaaaaatgtaacattcttttttatatataaataattggtggcatcacaaatagcca 2521 aagcagggtggagagagtgatccttcctgggtgcaggcaagaaggggatatgttttctac 2581 agagttttcaaaacagtgataaagctgtctacaagtcattgtgctttttatcatcactat 2641 gcccagacaatgtgaaacatcagagatgaagtgctcttcccacagaggtggactgatcct 2701 tctccccactcccttggtgtgtctctgaatgcaatgttgtcttggaaaacagctttccaa 2761 gcatttcactcctgagcacttgccagtttcctcacttgttcttcacatatccaggcaaag 2821 acatcctgtttgctatatgaagcattgtatcccgtataaaaggaaggaaagagagaaata 2881 tatttttacactcatcactcctcaggggctgtacaatcatgtagaaattgtttaatgtgc 2941 ctgtcaaatagccaaagagtgttaaaccctgagttcccacccatgtgtgtggtatggtta 3001 ggattcatccagatacacagagagaggcacaacaggaggagaaaggataggggtgtgggg 3061 acagcgggcccccaatatggtgtaatcgtggcaggtctctgcctgaagtgctatgtgggg 3121 tttttcttgttttaattttgactttaacccctgatttgtaagtttttcataaaataaaca 3181 gaatcataactcatgtagatggctataagtgccgtagtgttctgtgggtctctggtgtct 3241 gccagtgataagtgtggcaccccaggaaggctgtggaccccatcaaggtgctatgtgagg 3301 gccatgcttggggtggtggtgggcccagtagaccctgcagccatccatccagcctgccca 3361 ctcacactgcccttgtgtactcctgctttgctacgttatcattgatcaatgtccctggtt 3421 acctatgtgtttgaattatcttcgtgttacaggtgtttaatgattttgctccttctagct 3481 tatttgtatttcacctgtttttctttaaatcaacatggttacactctgtttcagcaactg 3541 tataaattaaacacaaattattactactgctaaaaaaaaaaaaaaaaaa

[0624] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18Ra, IL-1RL2, or IL1RL1 protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0625] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein can be designed using the software available at the Integrated DNA Technologies website.

[0626] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0627] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0628] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0629] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, -units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0630] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein (e.g., specificity for an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA, e.g., specificity for any one of SEQ ID NOs: 62-102). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA can be designed based upon the nucleotide sequence of any of the IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL-1, IL-1, IL-18, IL-36, IL-360, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, a SMAD7 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0631] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of an IL-1, IL-1, I-18, IL,-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL-1, IL-1, IL-18, IL-36, IL-360, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0632] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Natl. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0633] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0634] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0635] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Nat. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Nat. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res., 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0636] Another means by which expression of an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0637] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0638] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0639] The siRNA molecules used to decrease expression of an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0640] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing an IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 62-102, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0641] As described herein, inhibitory nucleic acids preferentially bind (e.g., hybridize) to a nucleic acid encoding IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein to treat allergic diseases (e.g., asthma (Corren et al., N. Engl. J. Med. 365: 1088-1098, 2011)), radiation lung injury (Chung et al., Sci. Rep. 6: 39714, 2016), ulcerative colitis (Hua et al., Br. J. Clin. Pharmacol. 80:101-109, 2015), dermatitis (Guttman-Yassky et al., Exp. Opin. Biol. Ther. 13(4):1517, 2013), and chronic obstructive pulmonary disease (COPD) (Walsh et al. (2010) Curr. Opin. Investig Drugs. 11(11):1305-1312, 2010).

[0642] Exemplary IL-1 inhibitors that are antisense nucleic acids are described in Yilmaz-Elis et al., Mol. Ther. Nucleic Acids 2(1): e66, 2013; Lu et al., J. Immunol. 190(12): 6570-6578, 2013), small interfering RNA (siRNA) (e.g., Ma et al., Ann. Hepatol. 15(2): 260-270, 2016), or combinations thereof. In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1 protein can be administered to a subject (e.g., a human subject) in need thereof.

[0643] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0644] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0645] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0646] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-1R1, IL1RAP, IL-18R, IL-1RL2, or IL1RL1) with a T.sub.m of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive).

[0647] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0648] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0649] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0650] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0651] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0652] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0653] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0654] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0655] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0656] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0657] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0658] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0659] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0660] In some embodiments, the IL-1 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, and IL-33. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to one or both of IL-1R1 and IL1RAP. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to IL-18R. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to one or both of IL1RL1 and IL1RAP. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind to one or both of IL-1RL2 and IL-1RAP.

[0661] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv).sub.2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0662] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0663] In some embodiments, the IL-1 inhibitor is canakinumab (ACZ885, Ilaris (Dhimolea, MAbs 2(1): 3-13, 2010; Yokota et al., Clin. Exp. Rheumatol. 2016; Torene et al., Ann. Rheum. Dis. 76(1):303-309, 2017; Gram, Curr. Opin. Chem. Biol. 32:1-9, 2016; Kontzias et al., Semin. Arthritis Rheum 42(2):201-205, 2012). In some embodiments, the IL-1 inhibitor is anakinra (Kineret; Beynon et al., J. Clin. Rheumatol. 23(3):181-183, 2017; Stanam et al., Oncotarget 7(46):76087-76100, 2016; Nayki et al., J. Obstet Gynaecol. Res. 42(11):1525-1533, 2016; Greenhalgh et al., Dis. ModelMech. 5(6):823-833, 2012), or a variant thereof. In some embodiments, the IL-1 inhibitor is gevokizumab (XOMA 052; Knicklebein et al., Am. J. Ophthalmol. 172:104-110, 2016; Roubille et al., Atherosclerosis 236(2):277-285, 2014; Issafras et al., J. Pharmacol. Exp. Ther. 348(1):202-215, 2014; Handa et al., Obesity 21(2):306-309, 2013; Geiler et al., Curr. Opin. Mol. Ther. 12(6):755-769, 2010), LY2189102 (Bihorel et al., AAPSJ. 16(5):1009-1117, 2014; Sloan-Lancaster et al., Diabetes Care 36(8):2239-2246, 2013), MABp1 (Hickish et al., Lancey Oncol. 18(2):192-201, 2017; Timper et al., J. Diabetes Complications 29(7):955-960, 2015), CDP-484 (Braddock et al., Drug Discov. 3:330-339, 2004), or a variant thereof (Dinarello et al., Nat. Rev. Drug Discov. 11(8): 633-652, 2012).

[0664] Further teachings of IL-1 inhibitors that are antibodies or antigen-binding fragments thereof are described in U.S. Pat. Nos. 5,075,222; 7,446,175; 7,531,166; 7,744,865; 7,829,093; and 8,273,350; US 2016/0326243; US 2016/0194392, and US 2009/0191187, each of which is incorporated by reference in its entirety.

[0665] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0666] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 1105 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 1105 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9 M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 1105 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0667] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0668] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1(inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1(inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1(inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins or Soluble Receptors

[0669] In some embodiments, the IL-1 inhibitor is a fusion protein or a soluble receptor. For example, a fusion can include an extracellular domain of any one of IL-1R1, IL1RAP, IL-18Ra, IL-1RL2, and IL1RL1 fused to a partner amino acid sequence (e.g., a stabilizing domain, e.g., an IgG Fc region, e.g., a human IgG Fc region). In some embodiments, the IL-1 inhibitor is a soluble version of one or both of IL-1RL1 and IL1RAP. In some embodiments, the IL-1 inhibitor is a soluble version of IL-18R. In some embodiments, the IL-1 inhibitor is a soluble version of one or both of IL-1RL2 and IL-1RAP.

[0670] In some embodiments, the IL-1 inhibitor is a fusion protein comprising or consisting of rilonacept (IL-1 Trap, Arcalyst) (see, e.g., Kapur & Bonk, P T 34(3):138-141, 2009; Church et al., Biologics 2(4):733-742, 2008; McDermott, Drugs Today (Barc) 45(6):423-430, 2009). In some embodiments, the IL-1 inhibitor is a fusion protein that is chimeric (e.g., EBI-005 (Isunakinra) (Furfine et al., Invest. Ophthalmol. Vs. Sci. 53(14):2340-2340, 2012; Goldstein et al., Eye Contact Lens 41(3):145-155, 2015; Goldstein et al., Eye Contact Lens, 2016)).

[0671] In some embodiments, the IL-1 inhibitor is a soluble receptor that comprises or consists of sIL-1RI and/or sIL-1RII (Svenson et al., Eur. J. Immunol. 25(10): 2842-2850, 1995).

Endogenous IL-I Inhibitor Peptides

[0672] In some embodiments, the IL-1 inhibitor can be an endogenous ligand or an active fragment thereof, e.g., IL-1Ra or IL-36Ra. IL-1Ra is an endogenous soluble protein that decreases the ability of IL-1 and IL-1 to bind to their receptor (e.g., a complex of IL-1R1 and IL1RAP proteins). IL-36Ra is an endogenous soluble protein that decreases the ability of IL-36, IL-36, and IL-36 to bind to their receptor (e.g., a complex of IL-1RL2 and IL-1RAP proteins). Exemplary sequences for IL-1Ra and IL-36Ra are shown below.

TABLE-US-00013 HumanIL-1RamRNATranscript1 (SEQIDNO:126) 1atttctttataaaccacaactctgggcccgcaatggcagtccactgccttgctgcagtca 61cagaatggaaatctgcagaggcctccgcagtcacctaatcactctcctcctcttcctgtt 121ccattcagagacgatctgccgaccctctgggagaaaatccagcaagatgcaagccttcag 181aatctgggatgttaaccagaagaccttctatctgaggaacaaccaactagttgctggata 241cttgcaaggaccaaatgtcaatttagaagaaaagatagatgtggtacccattgagcctca 301tgctctgttcttgggaatccatggagggaagatgtgcctgtcctgtgtcaagtctggtga 361tgagaccagactccagctggaggcagttaacatcactgacctgagcgagaacagaaagca 421ggacaagcgcttcgccttcatccgctcagacagtggccccaccaccagttttgagtctgc 481cgcctgccccggttggttcctctgcacagcgatggaagctgaccagcccgtcagcctcac 541caatatgcctgacgaaggcgtcatggtcaccaaattctacttccaggaggacgagtagta 601ctgcccaggcctgcctgttcccattcttgcatggcaaggactgcagggactgccagtccc 661cctgccccagggctcccggctatgggggcactgaggaccagccattgaggggtggaccct 721cagaaggcgtcacaacaacctggtcacaggactctgcctcctcttcaactgaccagcctc 781catgctgcctccagaatggtctttctaatgtgtgaatcagagcacagcagcccctgcaca 841aagcccttccatgtcgcctctgcattcaggatcaaaccccgaccacctgcccaacctgct 901ctcctcttgccactgcctcttcctccctcattccaccttcccatgccctggatccatcag 961gccacttgatgacccccaaccaagtggctcccacaccctgttttacaaaaaagaaaagac 1021cagtccatgagggaggtttttaagggtttgtggaaaatgaaaattaggatttcatgattt 1081ttttttttcagtccccgtgaaggagagcccttcatttggagattatgttctttcggggag 1141aggctgaggacttaaaatattcctgcatttgtgaaatgatggtgaaagtaagtggtagct 1201tttcccttctttttcttctttttttgtgatgtcccaacttgtaaaaattaaaagttatgg 1261tactatgttagccccataattttttttttccttttaaaacacttccataatctggactcc 1321tctgtccaggcactgctgcccagcctccaagctccatctccactccagattttttacagc 1381tgcctgcagtactttacctcctatcagaagtttctcagctcccaaggctctgagcaaatg 1441tggctcctgggggttctttcttcctctgctgaaggaataaattgctccttgacattgtag 1501agcttctggcacttggagacttgtatgaaagatggctgtgcctctgcctgtctcccccac 1561cgggctgggagctctgcagagcaggaaacatgactcgtatatgtctcaggtccctgcagg 1621gccaagcacctagcctcgctcttggcaggtactcagcgaatgaatgctgtatatgttggg 1681tgcaaagttccctacttcctgtgacttcagctctgttttacaataaaatcttgaaaatgc 1741ctaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa HumanIL-1RamRNATranscript2 (SEQIDNO:127) 1gggcagctccaccctgggagggactgtggcccaggtactgcccgggtgctactttatggg 61cagcagctcagttgagttagagtctggaagacctcagaagacctcctgtcctatgaggcc 121ctccccatggctttagctgacttgtatgaagaaggaggtggaggaggaggagaaggtgaa 181gacaatgctgactcaaaggagacgatctgccgaccctctgggagaaaatccagcaagatg 241caagccttcagaatctgggatgttaaccagaagaccttctatctgaggaacaaccaacta 301gttgctggatacttgcaaggaccaaatgtcaatttagaagaaaagatagatgtggtaccc 361attgagcctcatgctctgttcttgggaatccatggagggaagatgtgcctgtcctgtgtc 421aagtctggtgatgagaccagactccagctggaggcagttaacatcactgacctgagcgag 481aacagaaagcaggacaagcgcttcgccttcatccgctcagacagtggccccaccaccagt 541tttgagtctgccgcctgccccggttggttcctctgcacagcgatggaagctgaccagccc 601gtcagcctcaccaatatgcctgacgaaggcgtcatggtcaccaaattctacttccaggag 661gacgagtagtactgcccaggcctgcctgttcccattcttgcatggcaaggactgcaggga 721ctgccagtccccctgccccagggctcccggctatgggggcactgaggaccagccattgag 781gggtggaccctcagaaggcgtcacaacaacctggtcacaggactctgcctcctcttcaac 841tgaccagcctccatgctgcctccagaatggtctttctaatgtgtgaatcagagcacagca 901gcccctgcacaaagcccttccatgtcgcctctgcattcaggatcaaaccccgaccacctg 961cccaacctgctctcctcttgccactgcctcttcctccctcattccaccttcccatgccct 1021ggatccatcaggccacttgatgacccccaaccaagtggctcccacaccctgttttacaaa 1081aaagaaaagaccagtccatgagggaggtttttaagggtttgtggaaaatgaaaattagga 1141tttcatgatttttttttttcagtccccgtgaaggagagcccttcatttggagattatgtt 1201ctttcggggagaggctgaggacttaaaatattcctgcatttgtgaaatgatggtgaaagt 1261aagtggtagcttttcccttctttttcttctttttttgtgatgtcccaacttgtaaaaatt 1321aaaagttatggtactatgttagccccataattttttttttccttttaaaacacttccata 1381atctggactcctctgtccaggcactgctgcccagcctccaagctccatctccactccaga 1441ttttttacagctgcctgcagtactttacctcctatcagaagtttctcagctcccaaggct 1501ctgagcaaatgtggctcctgggggttctttcttcctctgctgaaggaataaattgctcct 1561tgacattgtagagcttctggcacttggagacttgtatgaaagatggctgtgcctctgcct 1621gtctcccccaccgggctgggagctctgcagagcaggaaacatgactcgtatatgtctcag 1681gtccctgcagggccaagcacctagcctcgctcttggcaggtactcagcgaatgaatgctg 1741tatatgttgggtgcaaagttccctacttcctgtgacttcagctctgttttacaataaaat 1801cttgaaaatgcctaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 1861aaaaa HumanIL-1RamRNATranscript3 (SEQIDNO:128) 1gggcagctccaccctgggagggactgtggcccaggtactgcccgggtgctactttatggg 61cagcagctcagttgagttagagtctggaagacctcagaagacctcctgtcctatgaggcc 121ctccccatggctttagagacgatctgccgaccctctgggagaaaatccagcaagatgcaa 181gccttcagaatctgggatgttaaccagaagaccttctatctgaggaacaaccaactagtt 241gctggatacttgcaaggaccaaatgtcaatttagaagaaaagatagatgtggtacccatt 301gagcctcatgctctgttcttgggaatccatggagggaagatgtgcctgtcctgtgtcaag 361tctggtgatgagaccagactccagctggaggcagttaacatcactgacctgagcgagaac 421agaaagcaggacaagcgcttcgccttcatccgctcagacagtggccccaccaccagtttt 481gagtctgccgcctgccccggttggttcctctgcacagcgatggaagctgaccagcccgtc 541agcctcaccaatatgcctgacgaaggcgtcatggtcaccaaattctacttccaggaggac 601gagtagtactgcccaggcctgcctgttcccattcttgcatggcaaggactgcagggactg 661ccagtccccctgccccagggctcccggctatgggggcactgaggaccagccattgagggg 721tggaccctcagaaggcgtcacaacaacctggtcacaggactctgcctcctcttcaactga 781ccagcctccatgctgcctccagaatggtctttctaatgtgtgaatcagagcacagcagcc 841cctgcacaaagcccttccatgtcgcctctgcattcaggatcaaaccccgaccacctgccc 901aacctgctctcctcttgccactgcctcttcctccctcattccaccttcccatgccctgga 961tccatcaggccacttgatgacccccaaccaagtggctcccacaccctgttttacaaaaaa 1021gaaaagaccagtccatgagggaggtttttaagggtttgtggaaaatgaaaattaggattt 1081catgatttttttttttcagtccccgtgaaggagagcccttcatttggagattatgttctt 1141tcggggagaggctgaggacttaaaatattcctgcatttgtgaaatgatggtgaaagtaag 1201tggtagcttttcccttctttttcttctttttttgtgatgtcccaacttgtaaaaattaaa 1261agttatggtactatgttagccccataattttttttttccttttaaaacacttccataatc 1321tggactcctctgtccaggcactgctgcccagcctccaagctccatctccactccagattt 1381tttacagctgcctgcagtactttacctcctatcagaagtttctcagctcccaaggctctg 1441agcaaatgtggctcctgggggttctttcttcctctgctgaaggaataaattgctccttga 1501cattgtagagcttctggcacttggagacttgtatgaaagatggctgtgcctctgcctgtc 1561tcccccaccgggctgggagctctgcagagcaggaaacatgactcgtatatgtctcaggtc 1621cctgcagggccaagcacctagcctcgctcttggcaggtactcagcgaatgaatgctgtat 1681atgttgggtgcaaagttccctacttcctgtgacttcagctctgttttacaataaaatctt 1741gaaaatgcctaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 1801aa HumanIL-1RamRNATranscript4 (SEQIDNO:129) 1gggcagctccaccctgggagggactgtggcccaggtactgcccgggtgctactttatggg 61cagcagctcagttgagttagagtctggaagacctcagaagacctcctgtcctatgaggcc 121ctccccatggctttagggggattataaaactaatcatcaaagccaagaaggcaagagcaa 181gcatgtaccgctgaaaacacaagataactgcataagtaatgactttcagtgcagattcat 241agctaacccataaactgctggggcaaaaatcatcttggaaggctctgaacctcagaaagg 301attcacaagacgatctgccgaccctctgggagaaaatccagcaagatgcaagccttcaga 361atctgggatgttaaccagaagaccttctatctgaggaacaaccaactagttgctggatac 421ttgcaaggaccaaatgtcaatttagaagaaaagatagatgtggtacccattgagcctcat 481gctctgttcttgggaatccatggagggaagatgtgcctgtcctgtgtcaagtctggtgat 541gagaccagactccagctggaggcagttaacatcactgacctgagcgagaacagaaagcag 601gacaagcgcttcgccttcatccgctcagacagtggccccaccaccagttttgagtctgcc 661gcctgccccggttggttcctctgcacagcgatggaagctgaccagcccgtcagcctcacc 721aatatgcctgacgaaggcgtcatggtcaccaaattctacttccaggaggacgagtagtac 781tgcccaggcctgcctgttcccattcttgcatggcaaggactgcagggactgccagtcccc 841ctgccccagggctcccggctatgggggcactgaggaccagccattgaggggtggaccctc 901agaaggcgtcacaacaacctggtcacaggactctgcctcctcttcaactgaccagcctcc 961atgctgcctccagaatggtctttctaatgtgtgaatcagagcacagcagcccctgcacaa 1021agcccttccatgtcgcctctgcattcaggatcaaaccccgaccacctgcccaacctgctc 1081tcctcttgccactgcctcttcctccctcattccaccttcccatgccctggatccatcagg 1141ccacttgatgacccccaaccaagtggctcccacaccctgttttacaaaaaagaaaagacc 1201agtccatgagggaggtttttaagggtttgtggaaaatgaaaattaggatttcatgatttt 1261tttttttcagtccccgtgaaggagagcccttcatttggagattatgttctttcggggaga 1321ggctgaggacttaaaatattcctgcatttgtgaaatgatggtgaaagtaagtggtagctt 1381ttcccttctttttcttctttttttgtgatgtcccaacttgtaaaaattaaaagttatggt 1441actatgttagccccataattttttttttccttttaaaacacttccataatctggactcct 1501ctgtccaggcactgctgcccagcctccaagctccatctccactccagattttttacagct 1561gcctgcagtactttacctcctatcagaagtttctcagctcccaaggctctgagcaaatgt 1621ggctcctgggggttctttcttcctctgctgaaggaataaattgctccttgacattgtaga 1681gcttctggcacttggagacttgtatgaaagatggctgtgcctctgcctgtctcccccacc 1741gggctgggagctctgcagagcaggaaacatgactcgtatatgtctcaggtccctgcaggg 1801ccaagcacctagcctcgctcttggcaggtactcagcgaatgaatgctgtatatgttgggt 1861gcaaagttccctacttcctgtgacttcagctctgttttacaataaaatcttgaaaatgcc 1921taaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa HumanIL-36RamRNAVariant1 (SEQIDNO:130) 1cgctgggaatcctgctcctcctcaggtcctggcagtttcagggcccctccctaggcctta 61cttaaaaggctgaggcatccttggaggaacaggcagactccacagctcccgccaggagaa 121aggaacattctgaggggagtctacaccctgtggagctcaagatggtcctgagtggggcgc 181tgtgcttccgaatgaaggactcggcattgaaggtgctttatctgcataataaccagcttc 241tagctggagggctgcatgcagggaaggtcattaaaggtgaagagatcagcgtggtcccca 301atcggtggctggatgccagcctgtcccccgtcatcctgggtgtccagggtggaagccagt 361gcctgtcatgtggggtggggcaggagccgactctaacactagagccagtgaacatcatgg 421agctctatcttggtgccaaggaatccaagagcttcaccttctaccggcgggacatggggc 481tcacctccagcttcgagtcggctgcctacccgggctggttcctgtgcacggtgcctgaag 541ccgatcagcctgtcagactcacccagcttcccgagaatggtggctggaatgcccccatca 601cagacttctacttccagcagtgtgactagggcaacgtgccccccagaactccctgggcag 661agccagctcgggtgaggggtgagtggaggagacccatggcggacaatcactctctctgct 721ctcaggacccccacgtctgacttagtgggcacctgaccactttgtcttctggttcccagt 781ttggataaattctgagatttggagctcagtccacggtcctcccccactggatggtgctac 841tgctgtggaatcttgtaaaaaccatgtggggtaaactgggaataacatgaaaagatttct 901gtggaggtggggtgggggagtggtgggaatcattcctgcttaatggtaactgaccagtgt 961taccctgagccccgcaggccaacccatccccagttgagccttatagggtcagtagctctc 1021cacatgaagacctgtcactcaccactatgcaggagagggaggtggtcatagagtcaggga 1081tctatggcccttggcccagccccacctccttccctttaatcctgccactgtcatatgcta 1141cctttcctatctcttccctcatcatcttgttgtgggcatgaggaggtgctgatgtcagaa 1201gaaatggctcgagctcagaagataaaagataagtagggtatgctgatcctcttttaaaaa 1261cccaagatacaatcaaaatcccagatgctggtctctattcccatgaaaaagtgctcatga 1321catattgagaagacctacttacaaagtggcatatattgcaatttattttaattaaaagat 1381acctatttatatatttctttatagaaaaaagtctggaagagtttacttcaattgtagcaa 1441tgtcagggtggtggcagtataggtgatttttcttttaattctgttaatttacctgtattt 1501cctaatttttctacaatgaagatgaattccttgtataaaaataagaaaagaaattaatct 1561tgaggtaagcagagtagacatcatctctgattgtcctcagcctccacttccccagagtaa 1621attcaaattgaatcgagctctgctgctctggttggttgtagtagtgatcaggaaacagat 1681ctcagcaaagccactgaggaggaggctgtgctgagtttgtgtggctggaatctctgggta 1741aggaacttaaagaacaaaaatcatctggtaattctttcctagaaggatcacagcccctgg 1801gattccaaggcattggatccagtctctaagaaggctgctgtactggttgaattgtgtccc 1861cctcaaattcacatccttcttggaatctcagtctgtgagtttatttggagataaggtctc 1921tgcagatgtagttagttaagacaaggtcatgctggatgaaggtagacctaaattcaatat 1981gactggtttccttgtatgaaaaggagaggacacagagacagaggagatgcggggaagact 2041atgtaaagatgaaggcagagatcggagttttgcagccacaagctaagaaacaccaaggat 2101tgtggcaaccatcagaagcttggaagaggcaaagaagaattcttccctagaggctttaga 2161gggataacggctctgctgaaaccttaatctcagacttccagcctcctgaacgaagaaaga 2221ataaatttcggctgttttaagccaccaaggataattggttacagcagctctaggaaacta 2281atacagctgctaaaatgatccctgtctcctcgtgtttacattctgtgtgtgtcccctccc 2341acaatgtaccaaagttgtctttgtgaccaatagaatatggcagaagtgatggcatgccac 2401ttccaagattaggttataaaagacactgcagcttctacttgagccctctctctctgccac 2461ccaccgcccccaatctatcttggctcactcgctctgggggaagctagctgccatgctatg 2521agcaggcctataaagagacttacgtggtaaaaaatgaagtctcctgcccacagccacatt 2581agtgaacctagaagcagagactctgtgagataatcgatgtttgttgttttaagttgctca 2641gttttggtctaacttgttatgcagcaatagataaataatatgcagagaaagagaaaaaaa 2701aaaaaaaaaaaaaaaaa HumanIL-36RamRNAVariant2 (SEQIDNO:131) 1ggagagtcccacctctaacatctcctgtaggcctggcaatggcaggcaggaaagacagag 61gaaggaaggagggagaagggaaggagtgaaggaaggagtgaaaaaggggagtctacaccc 121tgtggagctcaagatggtcctgagtggggcgctgtgcttccgaatgaaggactcggcatt 181gaaggtgctttatctgcataataaccagcttctagctggagggctgcatgcagggaaggt 241cattaaaggtgaagagatcagcgtggtccccaatcggtggctggatgccagcctgtcccc 301cgtcatcctgggtgtccagggtggaagccagtgcctgtcatgtggggtggggcaggagcc 361gactctaacactagagccagtgaacatcatggagctctatcttggtgccaaggaatccaa 421gagcttcaccttctaccggcgggacatggggctcacctccagcttcgagtcggctgccta 481cccgggctggttcctgtgcacggtgcctgaagccgatcagcctgtcagactcacccagct 541tcccgagaatggtggctggaatgcccccatcacagacttctacttccagcagtgtgacta 601gggcaacgtgccccccagaactccctgggcagagccagctcgggtgaggggtgagtggag 661gagacccatggcggacaatcactctctctgctctcaggacccccacgtctgacttagtgg 721gcacctgaccactttgtcttctggttcccagtttggataaattctgagatttggagctca 781gtccacggtcctcccccactggatggtgctactgctgtggaatcttgtaaaaaccatgtg 841gggtaaactgggaataacatgaaaagatttctgtggaggtggggtgggggagtggtggga 901atcattcctgcttaatggtaactgaccagtgttaccctgagccccgcaggccaacccatc 961cccagttgagccttatagggtcagtagctctccacatgaagacctgtcactcaccactat 1021gcaggagagggaggtggtcatagagtcagggatctatggcccttggcccagccccacctc 1081cttccctttaatcctgccactgtcatatgctacctttcctatctcttccctcatcatctt 1141gttgtgggcatgaggaggtgctgatgtcagaagaaatggctcgagctcagaagataaaag 1201ataagtagggtatgctgatcctcttttaaaaacccaagatacaatcaaaatcccagatgc 1261tggtctctattcccatgaaaaagtgctcatgacatattgagaagacctacttacaaagtg 1321gcatatattgcaatttattttaattaaaagatacctatttatatatttctttatagaaaa 1381aagtctggaagagtttacttcaattgtagcaatgtcagggtggtggcagtataggtgatt 1441tttcttttaattctgttaatttacctgtatttcctaatttttctacaatgaagatgaatt 1501ccttgtataaaaataagaaaagaaattaatcttgaggtaagcagagtagacatcatctct 1561gattgtcctcagcctccacttccccagagtaaattcaaattgaatcgagctctgctgctc 1621tggttggttgtagtagtgatcaggaaacagatctcagcaaagccactgaggaggaggctg 1681tgctgagtttgtgtggctggaatctctgggtaaggaacttaaagaacaaaaatcatctgg 1741taattctttcctagaaggatcacagcccctgggattccaaggcattggatccagtctcta 1801agaaggctgctgtactggttgaattgtgtccccctcaaattcacatccttcttggaatct 1861cagtctgtgagtttatttggagataaggtctctgcagatgtagttagttaagacaaggtc 1921atgctggatgaaggtagacctaaattcaatatgactggtttccttgtatgaaaaggagag 1981gacacagagacagaggagatgcggggaagactatgtaaagatgaaggcagagatcggagt 2041tttgcagccacaagctaagaaacaccaaggattgtggcaaccatcagaagcttggaagag 2101gcaaagaagaattcttccctagaggctttagagggataacggctctgctgaaaccttaat 2161ctcagacttccagcctcctgaacgaagaaagaataaatttcggctgttttaagccaccaa 2221ggataattggttacagcagctctaggaaactaatacagctgctaaaatgatccctgtctc 2281ctcgtgtttacattctgtgtgtgtcccctcccacaatgtaccaaagttgtctttgtgacc 2341aatagaatatggcagaagtgatggcatgccacttccaagattaggttataaaagacactg 2401cagcttctacttgagccctctctctctgccacccaccgcccccaatctatcttggctcac 2461tcgctctgggggaagctagctgccatgctatgagcaggcctataaagagacttacgtggt 2521aaaaaatgaagtctcctgcccacagccacattagtgaacctagaagcagagactctgtga 2581gataatcgatgtttgttgttttaagttgctcagttttggtctaacttgttatgcagcaat 2641agataaataatatgcagagaaagagaaaaaaaaaaaaaaaaaaaaaaaa

IL-13 Inhibitors

[0673] The term IL-13 inhibitor refers to an agent which decreases IL-13 expression and/or decreases the binding of IL-13 to an IL-13 receptor. In some embodiments, the IL-13 inhibitor decreases the ability of IL-13 to bind an IL-13 receptor (e.g., a complex including IL-4R and IL-13R1, or a complex including IL-13R1 and IL-13R2).

[0674] In some embodiments, the IL-13 inhibitor targets the IL-4R subunit. In some embodiments, the IL-13 inhibitor targets the IL-13R1. In some embodiments, the IL-13 inhibitor targets IL-13R2. In some embodiments, the IL-13 inhibitor targets an IL-13 receptor including IL-4R and IL-13R1. In some embodiments, the IL-13 inhibitor targets an IL-13 receptor including IL-13R1 and IL-13R2. In some embodiments, the IL-13 inhibitor targets IL-13.

[0675] In some embodiments, an IL-13 inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, or a fusion protein. In some embodiments, the inhibitory nucleic acid can be an antisense nucleic acid, a ribozyme, a small interfering RNA, a small hairpin RNA, or a microRNA. Examples of aspects of these different inhibitory nucleic acids are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of an IL-13, IL-13R1, IL-13R2, or IL-4Ra mRNA in a mammalian cell can be synthesized in vitro.

[0676] Inhibitory nucleic acids that can decrease the expression of IL-13, IL-13R1, IL-13R2, or IL-4Ra mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 132-138).

TABLE-US-00014 HumanIL-13mRNA (SEQIDNO:132) 1 aagccacccagcctatgcatccgctcctcaatcctctcctgttggcactgggcctcatgg 61 cgcttttgttgaccacggtcattgctctcacttgccttggcggctttgcctccccaggcc 121 ctgtgcctccctctacagccctcagggagctcattgaggagctggtcaacatcacccaga 181 accagaaggctccgctctgcaatggcagcatggtatggagcatcaacctgacagctggca 241 tgtactgtgcagccctggaatccctgatcaacgtgtcaggctgcagtgccatcgagaaga 301 cccagaggatgctgagcggattctgcccgcacaaggtctcagctgggcagttttccagct 361 tgcatgtccgagacaccaaaatcgaggtggcccagtttgtaaaggacctgctcttacatt 421 taaagaaactttttcgcgagggacagttcaactgaaacttcgaaagcatcattatttgca 481 gagacaggacctgactattgaagttgcagattcatttttctttctgatgtcaaaaatgtc 541 ttgggtaggcgggaaggagggttagggaggggtaaaattccttagcttagacctcagcct 601 gtgctgcccgtcttcagcctagccgacctcagccttccccttgcccagggctcagcctgg 661 tgggcctcctctgtccagggccctgagctcggtggacccagggatgacatgtccctacac 721 ccctcccctgccctagagcacactgtagcattacagtgggtgccccccttgccagacatg 781 tggtgggacagggacccacttcacacacaggcaactgaggcagacagcagctcaggcaca 841 cttcttcttggtcttatttattattgtgtgttatttaaatgagtgtgtttgtcaccgttg 901 gggattggggaagactgtggctgctagcacttggagccaagggttcagagactcagggcc 961 ccagcactaaagcagtggacaccaggagtccctggtaataagtactgtgtacagaattct 1021 gctacctcactggggtcctggggcctcggagcctcatccgaggcagggtcaggagagggg 1081 cagaacagccgctcctgtctgccagccagcagccagctctcagccaacgagtaatttatt 1141 gtttttccttgtatttaaatattaaatatgttagcaaagagttaatatatagaagggtac 1201 cttgaacactgggggaggggacattgaacaagttgtttcattgactatcaaactgaagcc 1261 agaaataaagttggtgacagat HumanIL-13R1mRNA (SEQIDNO:133) 1 tgccaaggctccagcccggccgggctccgaggcgagaggctgcatggagtggccggcgcg 61 gctctgcgggctgtgggcgctgctgctctgcgccggcggcgggggcgggggcgggggcgc 121 cgcgcctacggaaactcagccacctgtgacaaatttgagtgtctctgttgaaaacctctg 181 cacagtaatatggacatggaatccacccgagggagccagctcaaattgtagtctatggta 241 ttttagtcattttggcgacaaacaagataagaaaatagctccggaaactcgtcgttcaat 301 agaagtacccctgaatgagaggatttgtctgcaagtggggtcccagtgtagcaccaatga 361 gagtgagaagcctagcattttggttgaaaaatgcatctcacccccagaaggtgatcctga 421 gtctgctgtgactgagcttcaatgcatttggcacaacctgagctacatgaagtgttcttg 481 gctccctggaaggaataccagtcccgacactaactatactctctactattggcacagaag 541 cctggaaaaaattcatcaatgtgaaaacatctttagagaaggccaatactttggttgttc 601 ctttgatctgaccaaagtgaaggattccagttttgaacaacacagtgtccaaataatggt 661 caaggataatgcaggaaaaattaaaccatccttcaatatagtgcctttaacttcccgtgt 721 gaaacctgatcctccacatattaaaaacctctccttccacaatgatgacctatatgtgca 781 atgggagaatccacagaattttattagcagatgcctattttatgaagtagaagtcaataa 841 cagccaaactgagacacataatgttttctacgtccaagaggctaaatgtgagaatccaga 901 atttgagagaaatgtggagaatacatcttgtttcatggtccctggtgttcttcctgatac 961 tttgaacacagtcagaataagagtcaaaacaaataagttatgctatgaggatgacaaact 1021 ctggagtaattggagccaagaaatgagtataggtaagaagcgcaattccacactctacat 1081 aaccatgttactcattgttccagtcatcgtcgcaggtgcaatcatagtactcctgcttta 1141 cctaaaaaggctcaagattattatattccctccaattcctgatcctggcaagatttttaa 1201 agaaatgtttggagaccagaatgatgatactctgcactggaagaagtacgacatctatga 1261 gaagcaaaccaaggaggaaaccgactctgtagtgctgatagaaaacctgaagaaagcctc 1321 tcagtgatggagataatttatttttaccttcactgtgaccttgagaagattcttcccatt 1381 ctccatttgttatctgggaacttattaaatggaaactgaaactactgcaccatttaaaaa 1441 caggcagctcataagagccacaggtctttatgttgagtcgcgcaccgaaaaactaaaaat 1501 aatgggcgctttggagaagagtgtggagtcattctcattgaattataaaagccagcaggc 1561 ttcaaactaggggacaaagcaaaaagtgatgatagtggtggagttaatcttatcaagagt 1621 tgtgacaacttcctgagggatctatacttgctttgtgttctttgtgtcaacatgaacaaa 1681 ttttatttgtaggggaactcatttggggtgcaaatgctaatgtcaaacttgagtcacaaa 1741 gaacatgtagaaaacaaaatggataaaatctgatatgtattgtttgggatcctattgaac 1801 catgtttgtggctattaaaactcttttaacagtctgggctgggtccggtggctcacgcct 1861 gtaatcccagcaatttgggagtccgaggcgggcggatcactcgaggtcaggagttccaga 1921 ccagcctgaccaaaatggtgaaacctcctctctactaaaactacaaaaattaactgggtg 1981 tggtggcgcgtgcctgtaatcccagctactcgggaagctgaggcaggtgaattgtttgaa 2041 cctgggaggtggaggttgcagtgagcagagatcacaccactgcactctagcctgggtgac 2101 agagcaagactctgtctaaaaaacaaaacaaaacaaaacaaaacaaaaaaacctcttaat 2161 attctggagtcatcattcccttcgacagcattttcctctgctttgaaagccccagaaatc 2221 agtgttggccatgatgacaactacagaaaaaccagaggcagcttctttgccaagaccttt 2281 caaagccattttaggctgttaggggcagtggaggtagaatgactccttgggtattagagt 2341 ttcaaccatgaagtctctaacaatgtattttcttcacctctgctactcaagtagcattta 2401 ctgtgtctttggtttgtgctaggcccccgggtgtgaagcacagaccccttccaggggttt 2461 acagtctatttgagactcctcagttcttgccacttttttttttaatctccaccagtcatt 2521 tttcagaccttttaactcctcaattccaacactgatttccccttttgcattctccctcct 2581 tcccttccttgtagccttttgactttcattggaaattaggatgtaaatctgctcaggaga 2641 cctggaggagcagaggataattagcatctcaggttaagtgtgagtaatctgagaaacaat 2701 gactaattcttgcatattttgtaacttccatgtgagggttttcagcattgatatttgtgc 2761 attttctaaacagagatgaggtggtatcttcacgtagaacattggtattcgcttgagaaa 2821 aaaagaatagttgaacctatttctctttctttacaagatgggtccaggattcctcttttc 2881 tctgccataaatgattaattaaatagcttttgtgtcttacattggtagccagccagccaa 2941 ggctctgtttatgcttttggggggcatatattgggttccattctcacctatccacacaac 3001 atatccgtatatatcccctctactcttacttcccccaaatttaaagaagtatgggaaatg 3061 agaggcatttcccccaccccatttctctcctcacacacagactcatattactggtaggaa 3121 cttgagaactttatttccaagttgttcaaacatttaccaatcatattaatacaatgatgc 3181 tatttgcaattcctgctcctaggggaggggagataagaaaccctcactctctacaggttt 3241 gggtacaagtggcaacctgcttccatggccgtgtagaagcatggtgccctggcttctctg 3301 aggaagctggggttcatgacaatggcagatgtaaagttattcttgaagtcagattgaggc 3361 tgggagacagccgtagtagatgttctactttgttctgctgttctctagaaagaatatttg 3421 gttttcctgtataggaatgagattaattcctttccaggtattttataattctgggaagca 3481 aaacccatgcctccccctagccatttttactgttatcctatttagatggccatgaagagg 3541 atgctgtgaaattcccaacaaacattgatgctgacagtcatgcagtctgggagtggggaa 3601 gtgatcttttgttcccatcctcttcttttagcagtaaaatagctgagggaaaagggaggg 3661 aaaaggaagttatgggaatacctgtggtggttgtgatccctaggtcttgggagctcttgg 3721 aggtgtctgtatcagtggatttcccatcccctgtgggaaattagtaggctcatttactgt 3781 tttaggtctagcctatgtggattttttcctaacatacctaagcaaacccagtgtcaggat 3841 ggtaattcttattctttcgttcagttaagtttttcccttcatctgggcactgaagggata 3901 tgtgaaacaatgttaacatttttggtagtcttcaaccagggattgtttctgtttaacttc 3961 ttataggaaagcttgagtaaaataaatattgtctttttgtatgtca HumanIL-13R2mRNA (SEQIDNO:134) 1 gtaagaacactctcgtgagtctaacggtcttccggatgaaggctatttgaagtcgccata 61 acctggtcagaagtgtgcctgtcggcggggagagaggcaatatcaaggttttaaatctcg 121 gagaaatggctttcgtttgcttggctatcggatgcttatataccifictgataagcacaa 181 catttggctgtacttcatcttcagacaccgagataaaagttaaccctcctcaggattttg 241 agatagtggatcccggatacttaggttatctctatttgcaatggcaacccccactgtctc 301 tggatcattttaaggaatgcacagtggaatatgaactaaaataccgaaacattggtagtg 361 aaacatggaagaccatcattactaagaatctacattacaaagatgggtttgatcttaaca 421 agggcattgaagcgaagatacacacgcttttaccatggcaatgcacaaatggatcagaag 481 ttcaaagttcctgggcagaaactacttattggatatcaccacaaggaattccagaaacta 541 aagttcaggatatggattgcgtatattacaattggcaatatttactctgttcttggaaac 601 ctggcataggtgtacttcttgataccaattacaacttgttttactggtatgagggcttgg 661 atcatgcattacagtgtgttgattacatcaaggctgatggacaaaatataggatgcagat 721 ttccctatttggaggcatcagactataaagatttctatatttgtgttaatggatcatcag 781 agaacaagcctatcagatccagttatttcacttttcagcttcaaaatatagttaaacctt 841 tgccgccagtctatcttacttttactcgggagagttcatgtgaaattaagctgaaatgga 901 gcatacctttgggacctattccagcaaggtgttttgattatgaaattgagatcagagaag 961 atgatactaccttggtgactgctacagttgaaaatgaaacatacaccttgaaaacaacaa 1021 atgaaacccgacaattatgctttgtagtaagaagcaaagtgaatatttattgctcagatg 1081 acggaatttggagtgagtggagtgataaacaatgctgggaaggtgaagacctatcgaaga 1141 aaactttgctacgtttctggctaccatttggtttcatcttaatattagttatatttgtaa 1201 ccggtctgcttttgcgtaagccaaacacctacccaaaaatgattccagaatttttctgtg 1261 atacatgaagactttccatatcaagagacatggtattgactcaacagtttccagtcatgg 1321 ccaaatgttcaatatgagtctcaataaactgaatttttcttgcgaatgttgaaaaa HumanIL-4RmRNATranscriptVariant1 (SEQIDNO:135) 1 gggtctccgcgcccaggaaagccccgcgcggcgcgggccagggaagggccacccaggggt 61 cccccacttcccgcttgggcgcccggacggcgaatggagcaggggcgcgcagataattaa 121 agatttacacacagctggaagaaatcatagagaagccgggcgtggtggctcatgcctata 181 atcccagcacttttggaggctgaggcgggcagatcacttgagatcaggagttcgagacca 241 gcctggtgccttggcatctcccaatggggtggctttgctctgggctcctgttccctgtga 301 gctgcctggtcctgctgcaggtggcaagctctgggaacatgaaggtcttgcaggagccca 361 cctgcgtctccgactacatgagcatctctacttgcgagtggaagatgaatggtcccacca 421 attgcagcaccgagctccgcctgttgtaccagctggtttttctgctctccgaagcccaca 481 cgtgtatccctgagaacaacggaggcgcggggtgcgtgtgccacctgctcatggatgacg 541 tggtcagtgcggataactatacactggacctgtgggctgggcagcagctgctgtggaagg 601 gctccttcaagcccagcgagcatgtgaaacccagggccccaggaaacctgacagttcaca 661 ccaatgtctccgacactctgctgctgacctggagcaacccgtatccccctgacaattacc 721 tgtataatcatctcacctatgcagtcaacatttggagtgaaaacgacccggcagatttca 781 gaatctataacgtgacctacctagaaccctccctccgcatcgcagccagcaccctgaagt 841 ctgggatttcctacagggcacgggtgagggcctgggctcagtgctataacaccacctgga 901 gtgagtggagccccagcaccaagtggcacaactcctacagggagcccttcgagcagcacc 961 tcctgctgggcgtcagcgtttcctgcattgtcatcctggccgtctgcctgttgtgctatg 1021 tcagcatcaccaagattaagaaagaatggtgggatcagattcccaacccagcccgcagcc 1081 gcctcgtggctataataatccaggatgctcaggggtcacagtgggagaagcggtcccgag 1141 gccaggaaccagccaagtgcccacactggaagaattgtcttaccaagctcttgccctgtt 1201 ttctggagcacaacatgaaaagggatgaagatcctcacaaggctgccaaagagatgcctt 1261 tccagggctctggaaaatcagcatggtgcccagtggagatcagcaagacagtcctctggc 1321 cagagagcatcagcgtggtgcgatgtgtggagttgtttgaggccccggtggagtgtgagg 1381 aggaggaggaggtagaggaagaaaaagggagcttctgtgcatcgcctgagagcagcaggg 1441 atgacttccaggagggaagggagggcattgtggcccggctaacagagagcctgttcctgg 1501 acctgctcggagaggagaatgggggcttttgccagcaggacatgggggagtcatgccttc 1561 ttccaccttcgggaagtacgagtgctcacatgccctgggatgagttcccaagtgcagggc 1621 ccaaggaggcacctccctggggcaaggagcagcctctccacctggagccaagtcctcctg 1681 ccagcccgacccagagtccagacaacctgacttgcacagagacgcccctcgtcatcgcag 1741 gcaaccctgcttaccgcagcttcagcaactccctgagccagtcaccgtgtcccagagagc 1801 tgggtccagacccactgctggccagacacctggaggaagtagaacccgagatgccctgtg 1861 tcccccagctctctgagccaaccactgtgccccaacctgagccagaaacctgggagcaga 1921 tcctccgccgaaatgtcctccagcatggggcagctgcagcccccgtctcggcccccacca 1981 gtggctatcaggagtttgtacatgcggtggagcagggtggcacccaggccagtgcggtgg 2041 tgggcttgggtcccccaggagaggctggttacaaggccttctcaagcctgcttgccagca 2101 gtgctgtgtccccagagaaatgtgggtttggggctagcagtggggaagaggggtataagc 2161 ctttccaagacctcattcctggctgccctggggaccctgccccagtccctgtccccttgt 2221 tcacctttggactggacagggagccacctcgcagtccgcagagctcacatctcccaagca 2281 gctccccagagcacctgggtctggagccgggggaaaaggtagaggacatgccaaagcccc 2341 cacttccccaggagcaggccacagacccccttgtggacagcctgggcagtggcattgtct 2401 actcagcccttacctgccacctgtgcggccacctgaaacagtgtcatggccaggaggatg 2461 gtggccagacccctgtcatggccagtccttgctgtggctgctgctgtggagacaggtcct 2521 cgccccctacaacccccctgagggccccagacccctctccaggtggggttccactggagg 2581 ccagtctgtgtccggcctccctggcaccctcgggcatctcagagaagagtaaatcctcat 2641 catccttccatcctgcccctggcaatgctcagagctcaagccagacccccaaaatcgtga 2701 actttgtctccgtgggacccacatacatgagggtctcttaggtgcatgtcctcttgttgc 2761 tgagtctgcagatgaggactagggcttatccatgcctgggaaatgccacctcctggaagg 2821 cagccaggctggcagatttccaaaagacttgaagaaccatggtatgaaggtgattggccc 2881 cactgacgttggcctaacactgggctgcagagactggaccccgcccagcattgggctggg 2941 ctcgccacatcccatgagagtagagggcactgggtcgccgtgccccacggcaggcccctg 3001 caggaaaactgaggcccttgggcacctcgacttgtgaacgagttgttggctgctccctcc 3061 acagcttctgcagcagactgtccctgttgtaactgcccaaggcatgttttgcccaccaga 3121 tcatggcccacgtggaggcccacctgcctctgtctcactgaactagaagccgagcctaga 3181 aactaacacagccatcaagggaatgacttgggcggccttgggaaatcgatgagaaattga 3241 acttcagggagggtggtcattgcctagaggtgctcattcatttaacagagcttccttagg 3301 ttgatgctggaggcagaatcccggctgtcaaggggtgttcagttaaggggagcaacagag 3361 gacatgaaaaattgctatgactaaagcagggacaatttgctgccaaacacccatgcccag 3421 ctgtatggctgggggctcctcgtatgcatggaacccccagaataaatatgctcagccacc 3481 ctgtgggccgggcaatccagacagcaggcataaggcaccagttaccctgcatgttggccc 3541 agacctcaggtgctagggaaggcgggaaccttgggttgagtaatgctcgtctgtgtgttt 3601 tagtttcatcacctgttatctgtgtttgctgaggagagtggaacagaaggggtggagttt 3661 tgtataaataaagtttctttgtctctttaaaaaaaaaaaaaaaaaaaaaa HumanIL-4RmRNATranscriptVariant3 (SEQIDNO:136) 1 gggtctccgcgcccaggaaagccccgcgcggcgcgggccagggaagggccacccaggggt 61 cccccacttcccgcttgggcgcccggacggcgaatggagcaggggcgcgcaggtgccttg 121 gcatctcccaatggggtggctttgctctgggctcctgttccctgtgagctgcctggtcct 181 gctgcaggtggcaagctctgggaacatgaaggtcttgcaggagcccacctgcgtctccga 241 ctacatgagcatctctacttgcgagtggaagatgaatggtcccaccaattgcagcaccga 301 gctccgcctgttgtaccagctggtttttctgctctccgaagcccacacgtgtatccctga 361 gaacaacggaggcgcggggtgcgtgtgccacctgctcatggatgacgtggtcagtgcgga 421 taactatacactggacctgtgggctgggcagcagctgctgtggaagggctccttcaagcc 481 cagcgagcatgtgaaacccagggccccaggaaacctgacagttcacaccaatgtctccga 541 cactctgctgctgacctggagcaacccgtatccccctgacaattacctgtataatcatct 601 cacctatgcagtcaacatttggagtgaaaacgacccggcagatttcagaatctataacgt 661 gacctacctagaaccctccctccgcatcgcagccagcaccctgaagtctgggatttccta 721 cagggcacgggtgagggcctgggctcagtgctataacaccacctggagtgagtggagccc 781 cagcaccaagtggcacaactcctacagggagcccttcgagcagcacctcctgctgggcgt 841 cagcgtttcctgcattgtcatcctggccgtctgcctgttgtgctatgtcagcatcaccaa 901 gattaagaaagaatggtgggatcagattcccaacccagcccgcagccgcctcgtggctat 961 aataatccaggatgctcaggggtcacagtgggagaagcggtcccgaggccaggaaccagc 1021 caagtgcccacactggaagaattgtcttaccaagctcttgccctgttttctggagcacaa 1081 catgaaaagggatgaagatcctcacaaggctgccaaagagatgcctttccagggctctgg 1141 aaaatcagcatggtgcccagtggagatcagcaagacagtcctctggccagagagcatcag 1201 cgtggtgcgatgtgtggagttgtttgaggccccggtggagtgtgaggaggaggaggaggt 1261 agaggaagaaaaagggagcttctgtgcatcgcctgagagcagcagggatgacttccagga 1321 gggaagggagggcattgtggcccggctaacagagagcctgttcctggacctgctcggaga 1381 ggagaatgggggcttttgccagcaggacatgggggagtcatgccttcttccaccttcggg 1441 aagtacgagtgctcacatgccctgggatgagttcccaagtgcagggcccaaggaggcacc 1501 tccctggggcaaggagcagcctctccacctggagccaagtcctcctgccagcccgaccca 1561 gagtccagacaacctgacttgcacagagacgcccctcgtcatcgcaggcaaccctgctta 1621 ccgcagcttcagcaactccctgagccagtcaccgtgtcccagagagctgggtccagaccc 1681 actgctggccagacacctggaggaagtagaacccgagatgccctgtgtcccccagctctc 1741 tgagccaaccactgtgccccaacctgagccagaaacctgggagcagatcctccgccgaaa 1801 tgtcctccagcatggggcagctgcagcccccgtctcggcccccaccagtggctatcagga 1861 gtttgtacatgcggtggagcagggtggcacccaggccagtgcggtggtgggcttgggtcc 1921 cccaggagaggctggttacaaggccttctcaagcctgcttgccagcagtgctgtgtcccc 1981 agagaaatgtgggtttggggctagcagtggggaagaggggtataagcctttccaagacct 2041 cattcctggctgccctggggaccctgccccagtccctgtccccttgttcacctttggact 2101 ggacagggagccacctcgcagtccgcagagctcacatctcccaagcagctccccagagca 2161 cctgggtctggagccgggggaaaaggtagaggacatgccaaagcccccacttccccagga 2221 gcaggccacagacccccttgtggacagcctgggcagtggcattgtctactcagcccttac 2281 ctgccacctgtgcggccacctgaaacagtgtcatggccaggaggatggtggccagacccc 2341 tgtcatggccagtccttgctgtggctgctgctgtggagacaggtcctcgccccctacaac 2401 ccccctgagggccccagacccctctccaggtggggttccactggaggccagtctgtgtcc 2461 ggcctccctggcaccctcgggcatctcagagaagagtaaatcctcatcatccttccatcc 2521 tgcccctggcaatgctcagagctcaagccagacccccaaaatcgtgaactttgtctccgt 2581 gggacccacatacatgagggtctcttaggtgcatgtcctcttgttgctgagtctgcagat 2641 gaggactagggcttatccatgcctgggaaatgccacctcctggaaggcagccaggctggc 2701 agatttccaaaagacttgaagaaccatggtatgaaggtgattggccccactgacgttggc 2761 ctaacactgggctgcagagactggaccccgcccagcattgggctgggctcgccacatccc 2821 atgagagtagagggcactgggtcgccgtgccccacggcaggcccctgcaggaaaactgag 2881 gcccttgggcacctcgacttgtgaacgagttgttggctgctccctccacagcttctgcag 2941 cagactgtccctgttgtaactgcccaaggcatgttttgcccaccagatcatggcccacgt 3001 ggaggcccacctgcctctgtctcactgaactagaagccgagcctagaaactaacacagcc 3061 atcaagggaatgacttgggcggccttgggaaatcgatgagaaattgaacttcagggaggg 3121 tggtcattgcctagaggtgctcattcatttaacagagcttccttaggttgatgctggagg 3181 cagaatcccggctgtcaaggggtgttcagttaaggggagcaacagaggacatgaaaaatt 3241 gctatgactaaagcagggacaatttgctgccaaacacccatgcccagctgtatggctggg 3301 ggctcctcgtatgcatggaacccccagaataaatatgctcagccaccctgtgggccgggc 3361 aatccagacagcaggcataaggcaccagttaccctgcatgttggcccagacctcaggtgc 3421 tagggaaggcgggaaccttgggttgagtaatgctcgtctgtgtgttttagtttcatcacc 3481 tgttatctgtgtttgctgaggagagtggaacagaaggggtggagttttgtataaataaag 3541 tttctttgtctctttaaaaaaaaaaaaaaaaaaaaaa HumanIL-4RmRNATranscriptVariant4 (SEQIDNO:137) 1 gggtctccgcgcccaggaaagccccgcgcggcgcgggccagggaagggccacccaggggt 61 cccccacttcccgcttgggcgcccggacggcgaatggagcaggggcgcgcaggtgccttg 121 gcatctcccaatggggtggctttgctctgggctcctgttccctgtgagctgcctggtcct 181 gctgcaggtggcaagctctggactcttcaggatgccgtgtggagaaaggaagagggtgga 241 agccaggaggtctggagggaggtctggagtggaggagatgagaggctccggatccctctg 301 ggaggtagatttgaggacagattggaattgaggtgaaagacagagaaagagaagtggcca 361 ggatgactccaagatttctgacctaaactactgggaaggacgcggttgtcatttctgaaa 421 tgcagaaggatgccagaagagaagggaacatgaaggtcttgcaggagcccacctgcgtct 481 ccgactacatgagcatctctacttgcgagtggaagatgaatggtcccaccaattgcagca 541 ccgagctccgcctgttgtaccagctggtttttctgctctccgaagcccacacgtgtatcc 601 ctgagaacaacggaggcgcggggtgcgtgtgccacctgctcatggatgacgtggtcagtg 661 cggataactatacactggacctgtgggctgggcagcagctgctgtggaagggctccttca 721 agcccagcgagcatgtgaaacccagggccccaggaaacctgacagttcacaccaatgtct 781 ccgacactctgctgctgacctggagcaacccgtatccccctgacaattacctgtataatc 841 atctcacctatgcagtcaacatttggagtgaaaacgacccggcagatttcagaatctata 901 acgtgacctacctagaaccctccctccgcatcgcagccagcaccctgaagtctgggattt 961 cctacagggcacgggtgagggcctgggctcagtgctataacaccacctggagtgagtgga 1021 gccccagcaccaagtggcacaactcctacagggagcccttcgagcagcacctcctgctgg 1081 gcgtcagcgtttcctgcattgtcatcctggccgtctgcctgttgtgctatgtcagcatca 1141 ccaagattaagaaagaatggtgggatcagattcccaacccagcccgcagccgcctcgtgg 1201 ctataataatccaggatgctcaggggtcacagtgggagaagcggtcccgaggccaggaac 1261 cagccaagtgcccacactggaagaattgtcttaccaagctcttgccctgttttctggagc 1321 acaacatgaaaagggatgaagatcctcacaaggctgccaaagagatgcctttccagggct 1381 ctggaaaatcagcatggtgcccagtggagatcagcaagacagtcctctggccagagagca 1441 tcagcgtggtgcgatgtgtggagttgtttgaggccccggtggagtgtgaggaggaggagg 1501 aggtagaggaagaaaaagggagcttctgtgcatcgcctgagagcagcagggatgacttcc 1561 aggagggaagggagggcattgtggcccggctaacagagagcctgttcctggacctgctcg 1621 gagaggagaatgggggcttttgccagcaggacatgggggagtcatgccttcttccacctt 1681 cgggaagtacgagtgctcacatgccctgggatgagttcccaagtgcagggcccaaggagg 1741 cacctccctggggcaaggagcagcctctccacctggagccaagtcctcctgccagcccga 1801 cccagagtccagacaacctgacttgcacagagacgcccctcgtcatcgcaggcaaccctg 1861 cttaccgcagcttcagcaactccctgagccagtcaccgtgtcccagagagctgggtccag 1921 acccactgctggccagacacctggaggaagtagaacccgagatgccctgtgtcccccagc 1981 tctctgagccaaccactgtgccccaacctgagccagaaacctgggagcagatcctccgcc 2041 gaaatgtcctccagcatggggcagctgcagcccccgtctcggcccccaccagtggctatc 2101 aggagtttgtacatgcggtggagcagggtggcacccaggccagtgcggtggtgggcttgg 2161 gtcccccaggagaggctggttacaaggccttctcaagcctgcttgccagcagtgctgtgt 2221 ccccagagaaatgtgggtttggggctagcagtggggaagaggggtataagcctttccaag 2281 acctcattcctggctgccctggggaccctgccccagtccctgtccccttgttcacctttg 2341 gactggacagggagccacctcgcagtccgcagagctcacatctcccaagcagctccccag 2401 agcacctgggtctggagccgggggaaaaggtagaggacatgccaaagcccccacttcccc 2461 aggagcaggccacagacccccttgtggacagcctgggcagtggcattgtctactcagccc 2521 ttacctgccacctgtgcggccacctgaaacagtgtcatggccaggaggatggtggccaga 2581 cccctgtcatggccagtccttgctgtggctgctgctgtggagacaggtcctcgcccccta 2641 caacccccctgagggccccagacccctctccaggtggggttccactggaggccagtctgt 2701 gtccggcctccctggcaccctcgggcatctcagagaagagtaaatcctcatcatccttcc 2761 atcctgcccctggcaatgctcagagctcaagccagacccccaaaatcgtgaactttgtct 2821 ccgtgggacccacatacatgagggtctcttaggtgcatgtcctcttgttgctgagtctgc 2881 agatgaggactagggcttatccatgcctgggaaatgccacctcctggaaggcagccaggc 2941 tggcagatttccaaaagacttgaagaaccatggtatgaaggtgattggccccactgacgt 3001 tggcctaacactgggctgcagagactggaccccgcccagcattgggctgggctcgccaca 3061 tcccatgagagtagagggcactgggtcgccgtgccccacggcaggcccctgcaggaaaac 3121 tgaggcccttgggcacctcgacttgtgaacgagttgttggctgctccctccacagcttct 3181 gcagcagactgtccctgttgtaactgcccaaggcatgttttgcccaccagatcatggccc 3241 acgtggaggcccacctgcctctgtctcactgaactagaagccgagcctagaaactaacac 3301 agccatcaagggaatgacttgggcggccttgggaaatcgatgagaaattgaacttcaggg 3361 agggtggtcattgcctagaggtgctcattcatttaacagagcttccttaggttgatgctg 3421 gaggcagaatcccggctgtcaaggggtgttcagttaaggggagcaacagaggacatgaaa 3481 aattgctatgactaaagcagggacaatttgctgccaaacacccatgcccagctgtatggc 3541 tgggggctcctcgtatgcatggaacccccagaataaatatgctcagccaccctgtgggcc 3601 gggcaatccagacagcaggcataaggcaccagttaccctgcatgttggcccagacctcag 3661 gtgctagggaaggcgggaaccttgggttgagtaatgctcgtctgtgtgttttagtttcat 3721 cacctgttatctgtgtttgctgaggagagtggaacagaaggggtggagttttgtataaat 3781 aaagtttctttgtctctttaaaaaaaaaaaaaaaaaaaaaa HumanIL-4RmRNATranscriptVariant5 (SEQIDNO:138) 1 gggtctccgcgcccaggaaagccccgcgcggcgcgggccagggaagggccacccaggggt 61 cccccacttcccgcttgggcgcccggacggcgaatggagcaggggcgcgcaggtgccttg 121 gcatctcccaatggggtggctttgctctgggctcctgttccctgtgagctgcctggtcct 181 gctgcaggtggcaagctctgggaacatgaaggtcttgcaggagcccacctgcgtctccga 241 ctacatgagcatctctacttgcgagtggaagatgaatggtcccaccaattgcagcaccga 301 gctccgcctgttgtaccagctggtttttctgctctccgaagcccacacgtgtatccctga 361 gaacaacggaggcgcggggtgcgtgtgccacctgctcatggatgacgtggtcagtgcgga 421 taactatacactggacctgtgggctgggcagcagctgctgtggaagggctccttcaagcc 481 cagcgagcatgtgaaacccagggccccaggaaacctgacagttcacaccaatgtctccga 541 cactctgctgctgacctggagcaacccgtatccccctgacaattacctgtataatcatct 601 cacctatgcagtcaacatttggagtgaaaacgacccggcagataatctataacgtgacct 661 acctagaaccctccctccgcatcgcagccagcaccctgaagtctgggatttcctacaggg 721 cacgggtgagggcctgggctcagtgctataacaccacctggagtgagtggagccccagca 781 ccaagtggcacaactcctacagggagcccttcgagcagcacctcctgctgggcgtcagcg 841 tttcctgcattgtcatcctggccgtctgcctgttgtgctatgtcagcatcaccaagatta 901 agaaagaatggtgggatcagattcccaacccagcccgcagccgcctcgtggctataataa 961 tccaggatgctcaggggtcacagtgggagaagcggtcccgaggccaggaaccagccaagt 1021 gcccacactggaagaattgtcttaccaagctcttgccctgttttctggagcacaacatga 1081 aaagggatgaagatcctcacaaggctgccaaagagatgcctttccagggctctggaaaat 1141 cagcatggtgcccagtggagatcagcaagacagtcctctggccagagagcatcagcgtgg 1201 tgcgatgtgtggagttgtttgaggccccggtggagtgtgaggaggaggaggaggtagagg 1261 aagaaaaagggagcttctgtgcatcgcctgagagcagcagggatgacttccaggagggaa 1321 gggagggcattgtggcccggctaacagagagcctgttcctggacctgctcggagaggaga 1381 atgggggcttttgccagcaggacatgggggagtcatgccttcttccaccttcgggaagta 1441 cgagtgctcacatgccctgggatgagttcccaagtgcagggcccaaggaggcacctccct 1501 ggggcaaggagcagcctctccacctggagccaagtcctcctgccagcccgacccagagtc 1561 cagacaacctgacttgcacagagacgcccctcgtcatcgcaggcaaccctgcttaccgca 1621 gcttcagcaactccctgagccagtcaccgtgtcccagagagctgggtccagacccactgc 1681 tggccagacacctggaggaagtagaacccgagatgccctgtgtcccccagctctctgagc 1741 caaccactgtgccccaacctgagccagaaacctgggagcagatcctccgccgaaatgtcc 1801 tccagcatggggcagctgcagcccccgtctcggcccccaccagtggctatcaggagtttg 1861 tacatgcggtggagcagggtggcacccaggccagtgcggtggtgggcttgggtcccccag 1921 gagaggctggttacaaggccttctcaagcctgcttgccagcagtgctgtgtccccagaga 1981 aatgtgggtttggggctagcagtggggaagaggggtataagcctttccaagacctcattc 2041 ctggctgccctggggaccctgccccagtccctgtccccttgttcacctttggactggaca 2101 gggagccacctcgcagtccgcagagctcacatctcccaagcagctccccagagcacctgg 2161 gtctggagccgggggaaaaggtagaggacatgccaaagcccccacttccccaggagcagg 2221 ccacagacccccttgtggacagcctgggcagtggcattgtctactcagcccttacctgcc 2281 acctgtgcggccacctgaaacagtgtcatggccaggaggatggtggccagacccctgtca 2341 tggccagtccttgctgtggctgctgctgtggagacaggtcctcgccccctacaacccccc 2401 tgagggccccagacccctctccaggtggggttccactggaggccagtctgtgtccggcct 2461 ccctggcaccctcgggcatctcagagaagagtaaatcctcatcatccttccatcctgccc 2521 ctggcaatgctcagagctcaagccagacccccaaaatcgtgaactttgtctccgtgggac 2581 ccacatacatgagggtctcttaggtgcatgtcctcttgttgctgagtctgcagatgagga 2641 ctagggcttatccatgcctgggaaatgccacctcctggaaggcagccaggctggcagatt 2701 tccaaaagacttgaagaaccatggtatgaaggtgattggccccactgacgttggcctaac 2761 actgggctgcagagactggaccccgcccagcattgggctgggctcgccacatcccatgag 2821 agtagagggcactgggtcgccgtgccccacggcaggcccctgcaggaaaactgaggccct 2881 tgggcacctcgacttgtgaacgagttgttggctgctccctccacagcttctgcagcagac 2941 tgtccctgttgtaactgcccaaggcatgttttgcccaccagatcatggcccacgtggagg 3001 cccacctgcctctgtctcactgaactagaagccgagcctagaaactaacacagccatcaa 3061 gggaatgacttgggcggccttgggaaatcgatgagaaattgaacttcagggagggtggtc 3121 attgcctagaggtgctcattcatttaacagagcttccttaggttgatgctggaggcagaa 3181 tcccggctgtcaaggggtgttcagttaaggggagcaacagaggacatgaaaaattgctat 3241 gactaaagcagggacaatttgctgccaaacacccatgcccagctgtatggctgggggctc 3301 ctcgtatgcatggaacccccagaataaatatgctcagccaccctgtgggccgggcaatcc 3361 agacagcaggcataaggcaccagttaccctgcatgttggcccagacctcaggtgctaggg 3421 aaggcgggaaccttgggttgagtaatgctcgtctgtgtgttttagtttcatcacctgtta 3481 tctgtgtttgctgaggagagtggaacagaaggggtggagttttgtataaataaagtttct 3541 ttgtctctttaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa

[0677] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL-13, IL-13R1, IL-13R2, or IL-4R protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0678] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL-13, IL-13R1, IL-13R2, or IL-4R protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL-13, IL-13R1, IL-13R2, or IL-4R protein can be designed using the software available at the Integrated DNA Technologies website.

[0679] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0680] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0681] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL-13, IL-13R1, IL-13R2, or IL-4R protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0682] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 0-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0683] Non-limiting examples of IL-13 inhibitors that are antisense nucleic acids are described in Kim et al., J. Gene Med. 11(1): 26-37, 2009; and Mousavi et al., Iran J. Allergy Asthma Immunol. 2(3): 131-137, 2003.

[0684] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL-13, IL-13R1, IL-13R2, or IL-4R protein (e.g., specificity for an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA, e.g., specificity for any one of SEQ ID NOs: 109-115). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA can be designed based upon the nucleotide sequence of any of the IL-13, IL-13R1, IL-13R2, or IL-4R mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0685] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of an IL-13, IL-13R1, IL-13R2, or IL-4R polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL-13, IL-13R1, IL-13R2, or IL-4R polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0686] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Nat. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0687] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0688] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0689] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Nat. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Nat. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res., 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0690] Another means by which expression of an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL-13, IL-13R1, IL-13R2, or IL-4R polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0691] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0692] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0693] The siRNA molecules used to decrease expression of an IL-13, IL-13R1, IL-13R2, or IL-4Ra mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0694] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing an IL-13, IL-13R1, IL-13R2, or IL-4R mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 109-115, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0695] As described herein, inhibitory nucleic acids preferentially bind (e.g., hybridize) to a nucleic acid encoding IL-13, IL-13R1, IL-13R2, or IL-4R protein to treat allergic diseases (e.g., asthma (Corren et al., N. Engl. J Med. 365: 1088-1098, 2011)), radiation lung injury (Chung et al., Sci. Rep. 6: 39714, 2016), ulcerative colitis (Hua et al., Br. J. Clin. Pharmacol. 80:101-109, 2015), dermatitis (Guttman-Yassky et al., Exp. Opin. Biol. Ther. 13(4):1517, 2013), and chronic obstructive pulmonary disease (COPD) (Walsh et al. (2010) Curr. Opin. Investig Drugs. 11(11):1305-1312, 2010).

[0696] Non-limiting examples of short interfering RNA (siRNA) that are IL-13 inhibitors are described in Lively et al., J. Allergy Clin. Immunol. 121(1):88-94, 2008). Non-limiting examples of short hairpin RNA (shRNA) that are IL-13 inhibitors are described in Lee et al., Hum Gene Ther. 22(5):577-586, 2011, and Shilovskiy et al., Eur. Resp. J. 42:P523, 2013).

[0697] In some embodiments, an inhibitory nucleic acid can be a microRNA. Non-limiting examples of microRNAs that are IL-13 inhibitors are let-7 (Kumar et al., J. Allergy Clin. Immunol. 128(5):1077-1085, 2011).

[0698] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a IL-13, IL-13R1, IL-13R2, or IL-4R protein can be administered to a subject (e.g., a human subject) in need thereof.

[0699] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0700] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0701] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-13, IL-13R1, IL-13R2, or IL-4Ra) with a Tm of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0702] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-13, IL-13R1, IL-13R2, or IL-4Ra) with a Tm of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive).

[0703] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0704] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0705] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0706] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0707] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0708] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0709] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0710] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0711] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0712] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0713] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0714] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0715] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies

[0716] In some embodiments, the IL-13 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of IL-13, IL-13R1, IL-13R2, or IL-4R, or a combination thereof. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to IL-13. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to an IL-13 receptor (e.g., a complex including IL-4R and IL-13R1, or a complex including IL-13R1 and IL-13R2).

[0717] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv)2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0718] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab)2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0719] In some embodiments, the IL-13 inhibitor is a monoclonal antibody (Bagnasco et al., Int. Arch. Allergy Immunol. 170:122-131, 2016). In some embodiments, the IL-13 inhibitor is QAX576 (Novartis) or an antigen-binding fragment thereof (see, e.g., Kariyawasam et al., B92 New Treatment Approachesfor Asthma and Allergery San Diego, 2009; Rothenberg et al., J. Allergy Clin. Immunol. 135:500-507, 2015). In some embodiments, the IL-13 inhibitor is ABT-308 (Abbott) or an antigen-binding fragment thereof (see, e.g., Ying et al., American Thoracic Society 2010 International Conference, May 14-19, 2010, New Orleans; Abstract A6644). In some embodiments, the IL-13 inhibitor is CNTO-5825 (Centrocore) or an antigen-binding fragment thereof (see, e.g., van Hartingsveldt et al., British J. Clin. Pharmacol. 75:1289-1298, 2013). In some embodiments, the IL-13 inhibitor is dupilumab (REGN668/SAR231893) or an antigen-binding fragment thereof (see, e.g., Simpson et al., N. Eng. J. Med. 375:2335-2348, 2016; Thaci et al., Lancet 387:40-52, 2016). In some embodiments, the IL-13 inhibitor is AMG317 (Amgen) or an antigen-binding fragment thereof (Polosa et al., Drug Discovery Today 17:591-599, 2012; Holgate, British J. Clinical Pharmacol. 76:277-291, 2013). In some embodiments, the IL-13 inhibitor is an antibody that specifically binds to IL-13R1 (see, e.g., U.S. Pat. No. 7,807,158; WO 96/29417; WO 97/15663; and WO 03/080675).

[0720] In some embodiments, the IL-13 inhibitor is a humanized monoclonal antibody (e.g., lebrikizumab (TNX-650) (Thomson et al., Biologics 6:329-335, 2012; and Hanania et al., Thorax 70(8):748-756, 2015). In some embodiments, the IL-13 inhibitor is an anti-IL-13 antibody, e.g., GSK679586 or a variant thereof (Hodsman et al., Br. J Clin. Pharmacol. 75(1):118-128, 2013; and De Boever et al., J. Allergy Clin. Immunol. 133(4):989-996, 2014). In some embodiments, the IL-13 inhibitor is tralokinumab (CAT-354) or a variant thereof (Brightling et al., Lancet 3(9): 692-701, 2015; Walsh et al. (2010) Curr. Opin. Investig. Drugs 11(11):1305-1312, 2010; Piper et al., Euro. Resp. J. 41:330-338, 2013; May et al., Br. J. Pharmacol. 166(1): 177-193, 2012; Singh et al., BMC Pulm Med. 10:3, 2010; Blanchard et al., Clin. Exp. Allergy 35(8): 1096-1103, 2005). In some embodiments, the Il-13 inhibitor is anrukinzumab (IMA-638) (Hua et al., Br. J Clin. Pharmacol. 80: 101-109, 2015; Reinisch et al., Gut 64(6): 894-900, 2015; Gauvreau et al., Am. J. Respir. Crit. Care Med. 183(8):1007-1014, 2011; Bree et al., J. Allergy Clin. Immunol. 119(5):1251-1257, 2007). Further teachings of IL-13 inhibitors that are antibodies or antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,067,199; 7,910,708; 8,221,752; 8,388,965; 8,399,630; and 8,734,801; US 2014/0341913; US 2015/0259411; US 2016/0075777; US 2016/0130339, US 2011/0243928, and US 2014/0105897 each of which is incorporated by reference in its entirety.

[0721] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0722] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9 M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 1109 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0723] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0724] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1(inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1(inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1(inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins

[0725] In some embodiments, the IL-13 inhibitor is a fusion protein or a soluble antagonist. In some embodiments, the fusion protein comprises a soluble fragment of a receptor of IL-13 (e.g., a soluble fragment of a complex including IL-13R1 and IL-4Ra, a soluble fragment of a complex including IL-13R1 and IL-13R2, a soluble fragment of IL-13R1, a soluble fragment of IL-13R2, or soluble fragment of IL-4R). In some embodiments, the fusion protein comprises an extracellular domain of a receptor of IL-13 (e.g., a fusion protein including an extracellular domain of both IL-13R1 and IL-4Ra, a fusion protein including an extracellular domain of both IL-13R1 and IL-13R2, a fusion protein including an extracellular domain of IL-13R1, a fusion protein including an extracellular domain of IL-13R2, or a fusion protein including an extracellular domain of IL-4R).

[0726] In some embodiments, the fusion protein comprises or consists of sIL-13R2-Fc (see, e.g., Chiaramonte et al., J. Clin. Invest. 104(6):777-785, 1999; Kasaian et al., Am. J. Respir. Cell. Mol. Biol. 36(3):368-376, 2007; Miyahara et al., J. Allergy Clin. Immunol. 118(5):1110-1116, 2006; Rahaman et al., Cancer Res. 62(4):1103-1109, 2002; incorporated by reference herein). In some embodiments, the fusion protein comprises or consists of an IL-13 fusion cytotoxin (e.g., IL-13/diphtheria toxin fusion protein (Li et al., Protein Eng. 15(5):419-427, 2002), IL-13-PE38QQR (IL-13-PE) (Blease et al. (2001) J. Immunol. 167(11):6583-6592, 2001; and Husain et al., J. Neuro-Oncol. 65(1):37-48, 2003)).

IL-10 Receptor Agonists

[0727] The term IL-10 receptor agonist is any molecule that binds to and activates a receptor for IL-10 expressed on a mammalian cell or a nucleic acid that encodes any such molecule. A receptor for IL-10 can include, e.g., a complex of two IL-10 receptor-1 (IL-10R1) proteins and two IL-10 receptor 2 (IL-10R2) proteins. In some examples, an IL-10 receptor agonist is an antibody or an antigen-binding antibody fragment that specifically binds to and activates a receptor for IL-10 (e.g., a human receptor for IL-10). In some examples, an IL-10 receptor agonist is a recombinant IL-10 (e.g., human recombinant IL-10). In some examples, an IL-10 receptor agonist is a pegylated recombinant IL-10 (e.g., pegylated recombinant human IL-10). In some examples, an IL-10 receptor agonist is a fusion protein. In some examples, an IL-10 receptor agonist is an IL-10 peptide mimetic.

[0728] In some embodiments, any of the devices or compositions described herein can contain a recombinant mammalian cell (e.g., a recombinant human cell) that secretes an IL-10 receptor agonist (e.g., a recombinant IL-10, e.g., a recombinant human IL-10). In some embodiments, any of the devices or compositions described herein can contain a mammalian cell (e.g., a human cell) that secretes IL-10 (e.g., human IL-10).

[0729] Activation of an IL-10 receptor in a mammalian cell can be determined by detecting an increase in the activation of downstream signaling proteins in a mammalian cell contacted with an IL-10 receptor agonist. For example, activation of an IL-10 receptor in a mammalian cell can be detected by an increase in the phosphorylation and activity of JAK1 and TYK2, phosphorylation and subsequent nuclear translocation of STAT3, and/or increased transcription of BCLXL, Cyclin-D1, Cyclin-D2, Cyclin-D3, Cyclin-A, Pim1, c-Myc, or p19 (INK4D) (see, e.g., Hu et al., J Leukoc. Biol. 82(2):237-243, 2007; and Cavalcante et al., J. Periodontol. 83(7):926-935, 2012). Reagents for detecting these downstream events that indicate activation of an IL-10 receptor are available from, e.g., ThermoFisher Scientific.

IL-10 and IL-10 Receptor

[0730] Exemplary sequences of human IL-10 proteins and cDNA sequences are shown below.

TABLE-US-00015 PrecursorHumanIL-10Protein(withsignalsequenceinbold) (SEQIDNO:139) 1 mhssallcclvlltgvraspgqgtqsenscthfpgnlpnmlrdlrdafsrvktffqmkdq 61 ldnlllkeslledfkgylgcqalsemiqfyleevmpqaenqdpdikahvnslgenlktlr 121 lrlrrchrflpcenkskaveqvknafnklqekgiykamsefdifinyieaymtmkirn MatureHumanIL-10Protein (SEQIDNO:140) spgqgtqsenscthfpgnlpnmlrdlrdafsrvktffqmkdqldnlllkesl ledfkgylgcqalsemiqfyleevmpqaenqdpdikahvnslgenlktlr lrlachrflpcenkskaveqvknafnklqekgiykamsefdifinyiea ymtmkirn HumanIL-10cDNA (SEQIDNO:141) 1 acacatcaggggcttgctcttgcaaaaccaaaccacaagacagacttgcaaaagaaggca 61 tgcacagctcagcactgctctgttgcctggtcctcctgactggggtgagggccagcccag 121 gccagggcacccagtctgagaacagctgcacccacttcccaggcaacctgcctaacatgc 181 ttcgagatctccgagatgccttcagcagagtgaagactttctttcaaatgaaggatcagc 241 tggacaacttgttgttaaaggagtccttgctggaggactttaagggttacctgggttgcc 301 aagccttgtctgagatgatccagttttacctggaggaggtgatgccccaagctgagaacc 361 aagacccagacatcaaggcgcatgtgaactccctgggggagaacctgaagaccctcaggc 421 tgaggctacggcgctgtcatcgatttcttccctgtgaaaacaagagcaaggccgtggagc 481 aggtgaagaatgcctttaataagctccaagagaaaggcatctacaaagccatgagtgagt 541 ttgacatcttcatcaactacatagaagcctacatgacaatgaagatacgaaactgagaca 601 tcagggtggcgactctatagactctaggacataaattagaggtctccaaaatcggatctg 661 gggctctgggatagctgacccagccccttgagaaaccttattgtacctctcttatagaat 721 atttattacctctgatacctcaacccccatttctatttatttactgagcttctctgtgaa 781 cgatttagaaagaagcccaatattataatttttttcaatatttattattttcacctgttt 841 ttaagctgtttccatagggtgacacactatggtatttgagtgttttaagataaattataa 901 gttacataagggaggaaaaaaaatgttctttggggagccaacagaagcttccattccaag 961 cctgaccacgctttctagctgttgagctgttttccctgacctccctctaatttatcttgt 1021 ctctgggcttggggcttcctaactgctacaaatactcttaggaagagaaaccagggagcc 1081 cctttgatgattaattcaccttccagtgtctcggagggattcccctaacctcattcccca 1141 accacttcattcttgaaagctgtggccagcttgttatttataacaacctaaatttggttc 1201 taggccgggcgcggtggctcacgcctgtaatcccagcactttgggaggctgaggcgggtg 1261 gatcacttgaggtcaggagttcctaaccagcctggtcaacatggtgaaaccccgtctcta 1321 ctaaaaatacaaaaattagccgggcatggtggcgcgcacctgtaatcccagctacttggg 1381 aggctgaggcaagagaattgcttgaacccaggagatggaagttgcagtgagctgatatca 1441 tgcccctgtactccagcctgggtgacagagcaagactctgtctcaaaaaataaaaataaa 1501 aataaatttggttctaatagaactcagttttaactagaatttattcaattcctctgggaa 1561 tgttacattgtttgtctgtcttcatagcagattttaattttgaataaataaatgtatctt 1621 attcacatc

[0731] The protein and cDNA sequences of exemplary non-human homologues of IL-10 are shown below.

TABLE-US-00016 PrecursorMouseIL-10Protein(withsignalsequenceinbold) (SEQIDNO:142) 1 mpgsallcclllltgmrisrgqysrednncthfpvgqshmllelrtafsqvktffqtkdq 61 ldnilltdslmqdfkgylgcqalsemiqfylvevmpqaekhgpeikehlnslgeklktlr 121 mrlrrchrflpcenkskaveqvksdfnklqdqgvykamnefdifincieaymmikmks MouseIL-10cDNA (SEQIDNO:143) 1 acatttagagacttgctcttgcactaccaaagccacaaggcagccttgcagaaaagagag 61 ctccatcatgcctggctcagcactgctatgctgcctgctcttactgactggcatgaggat 121 cagcaggggccagtacagccgggaagacaataactgcacccacttcccagtcggccagag 181 ccacatgctcctagagctgcggactgccttcagccaggtgaagactttctttcaaacaaa 241 ggaccagctggacaacatactgctaaccgactccttaatgcaggactttaagggttactt 301 gggttgccaagccttatcggaaatgatccagttttacctggtagaagtgatgccccaggc 361 agagaagcatggcccagaaatcaaggagcatttgaattccctgggtgagaagctgaagac 421 cctcaggatgcggctgaggcgctgtcatcgatttctcccctgtgaaaataagagcaaggc 481 agtggagcaggtgaagagtgattttaataagctccaagaccaaggtgtctacaaggccat 541 gaatgaatttgacatcttcatcaactgcatagaagcatacatgatgatcaaaatgaaaag 601 ctaaaacacctgcagtgtgtattgagtctgctggactccaggacctagacagagctctct 661 aaatctgatccagggatcttagctaacggaaacaactccttggaaaacctcgtttgtacc 721 tctctccgaaatatttattacctctgatacctcagttcccattctatttattcactgagc 781 ttctctgtgaactatttagaaagaagcccaatattataattttacagtatttattatttt 841 taacctgtgtttaagctgtttccattggggacactttatagtatttaaagggagattata 901 ttatatgatgggaggggttcttccttgggaagcaattgaagcttctattctaaggctggc 961 cacacttgagagctgcagggccctttgctatggtgtcctttcaattgctctcatccctga 1021 gttcagagctcctaagagagttgtgaagaaactcatgggtcttgggaagagaaaccaggg 1081 agatcctttgatgatcattcctgcagcagctcagagggttcccctactgtcatcccccag 1141 ccgcttcatccctgaaaactgtggccagtttgttatttataaccacctaaaattagttct 1201 aatagaactcatttttaactagaagtaatgcaattcctctgggaatggtgtattgtttgt 1261 ctgcctttgtagcagactctaattttgaataaatggatcttattcg PrecursorRatIL-10Protein(withsignalsequenceinbold) (SEQIDNO:144) 1 mpgsallccllllagvktskghsirgdnncthfpvsqthmlrelraafsqvktffqkkdq 61 ldnilltdsllqdfkgylgcqalsemikfylvevmpqaenhgpeikehlnslgeklktlw 121 iqlrrchrflpcenkskaveqvkndfnklqdkgvykamnefdifincieayvtlkmkn RatIL-10cDNA (SEQIDNO:145) 1 catgcctggctcagcactgctatgttgcctgctcttactggctggagtgaagaccagcaa 61 aggccattccatccggggtgacaataactgcacccacttcccagtcagccagacccacat 121 gctccgagagctgagggctgccttcagtcaagtgaagactttctttcaaaagaaggacca 181 gctggacaacatactgctgacagattccttactgcaggactttaagggttacttgggttg 241 ccaagccttgtcagaaatgatcaagttttacctggtagaagtgatgccccaggcagagaa 301 ccatggcccagaaatcaaggagcatttgaattccctgggagagaagctgaagaccctctg 361 gatacagctgcgacgctgtcatcgatttctcccctgtgagaataaaagcaaggcagtgga 421 gcaggtgaagaatgattttaataagctccaagacaaaggtgtctacaaggccatgaatga 481 gtttgacatcttcatcaactgcatagaagcctacgtgacactcaaaatgaaaaattgaac 541 cacccggcatctactggactgcaggacataaatagagcttctaaatctgatccagagatc 601 ttagctaacgggagcaactccttggaaaacctcgtttgtacctctctccaaaatatttat 661 tacctctgatacctcagttccc PrecursorRabbitIL-10Protein (SEQIDNO:146) 1 mlssallcclvflggtgasrgqdtpaenscihfpgglphmlrelraafgrvktffqskdq 61 lnsmllteslledlkgylgcqalsemiqfylkdvmpqaenhspairehvnslgenlktlr 121 lrlrqchrflpcenkskaveqvksafsklqeegvykamsefdifinyietymtmkiks RabbitIL-10cDNA (SEQIDNO:147) 1 aaagcaaaccacaaggcggactcgtagaagcaggcagagttccaccatgctcagctcagc 61 tctgctatgttgcctggtcttcctgggtgggacaggggccagccgaggccaggacacccc 121 tgctgagaacagctgcattcactttccaggcggcctgccccacatgctccgcgagctccg 181 tgctgcctttggcagggtgaagactttctttcaatcgaaggatcagctgaacagcatgtt 241 gttaaccgagtccctgctggaggaccttaagggttacctgggatgccaagccttgtcgga 301 gatgatccagttttacctgaaggacgtgatgccgcaagctgagaaccacagtccagccat 361 cagggagcacgtgaactccctgggggaaaacctgaagaccctcaggctgaggctgcgaca 421 atgtcaccgatttctcccctgtgaaaacaagagcaaggcagtggagcaggtgaagagcgc 481 cttcagcaagctgcaagaggaaggcgtctacaaagccatgagtgagtttgacatcttcat 541 caactacatagaaacctacatgacaatgaagataaaaagctaaaagccccaggatggcaa 601 ctcggctagagtctaggacatcagttagggacctgcacaccctgggtcagctgacccagc 661 accttggaaagctgttgtacctctcaatatttattacctctgatacctcagctcccgatc 721 ctatttatttaccgagcttctctgtgaactctttagaaagaagcccactattataatttt 781 ttcagtatttattattttcacctgcatttaagctgtacccatggggtgatgccctgtggg 841 atttgagtgtcttaggagaaattataatttatgtgaaagggaaaatgtgccttggggagc 901 cgactgaggcttccattccttctgtgcctgaccacactttctaactcctaagccgagctc 961 cctcttaccctctggagcccggacctgggtctcgagtgttccagagactcctagcctctt 1021 aggaagagagaccggaagcccttgggtggtgaccttccggcagctcagagggaggctcct 1081 gacctcgat PrecursorMonkeyIL-10Protein(withsignalsequenceinbold) (SEQIDNO:148) 1 mhssallcclvlltgvraspgqgtqsensctrfpgnlphmlrdlrdafsrvktffqmkdq 61 ldnillkeslledfkgylgcqalsemiqfyleevmpqaenhdpdikehvnslgenlktlr 121 lrlrrchrflpcenkskaveqvknafsklqekgvykamsefdifinyieaymtmkiqn MonkeyIL-10cDNA (SEQIDNO:149) 1 agaaggcatgcacagctcagcactgctctgttgcctagtcctcctgactggggtgagggc 61 cagcccaggccagggcacccagtctgagaacagctgcacccgcttcccaggcaacctgcc 121 tcacatgcttcgagacctccgagatgccttcagcagagtgaagactttctttcaaatgaa 181 ggatcagctggacaacatattgttaaaggagtccttgctggaggactttaagggttacct 241 gggttgccaagccttgtctgagatgatccagttttacctggaggaggtgatgccccaagc 301 tgagaaccacgacccagacatcaaggagcatgtgaactccctgggggagaatctgaagac 361 cctcaggctgaggctgcggcgctgtcatcgatttcttccctgtgaaaacaagagcaaggc 421 cgtggagcaggtgaagaatgcctttagtaagctccaagagaaaggcgtctacaaagccat 481 gagtgagtdgacatcttcatcaactacatagaagcctacatgacaatgaagatacaaaa 541 ctgagacatcagggtggcgactctatagactctaggacataaattagaggtctccaaaat 601 cagatccagggttctgggatagctgacccagccccttgagaaa

[0732] Exemplary protein and cDNA sequences for human IL-10R-1 and human IL-10R-2 are shown below.

TABLE-US-00017 PrecursorHumanIL-10R-1Protein(withsignalsequenceinbold) (SEQIDNO:150) 1 mlpclvvllaallslrlgsdahgtelpsppsvwfeaeffhhilhwtpipnqsestcyeva 61 llrygieswnsisncsqtlsydltavtldlyhsngyrarvravdgsrhsnwtvtntrfsv 121 devtltvgsvnleihngfilgkiqlprpkmapandtyesifshfreyeiairkvpgnftf 181 thkkvkhenfslltsgevgefcvqvkpsvasrsnkgmwskeecisltrqyftvtnviiff 241 afvlllsgalayclalqlyvrrrkklpsvllfkkpspfifisqrpspetqdtihpldeea 301 flkvspelknldlhgstdsgfgstkpslqteepqfllpdphpqadrtlgnreppvlgdsc 361 ssgssnstdsgiclqepslspstgptweqqvgsnsrgqddsgidlvqnsegragdtqggs 421 alghhsppepevpgeedpaavafqgylrqtrcaeekatktgcleeespltdglgpkfgrc 481 lvdeaglhppalakgylkqdplemtlassgaptgqwnqpteewsllalsscsdlgisdws 541 fahdlaplgcvaapggllgsfnsdlvtlplisslqsse HumanIL-10R-1cDNA,transcriptvariant1 (SEQIDNO:151) 1 gtcagtcccagcccaagggtagctggaggcgcgcaggccggctccgctccggccccggac 61 gatgcggcgcgcccaggatgctgccgtgcctcgtagtgctgctggcggcgctcctcagcc 121 tccgtcttggctcagacgctcatgggacagagctgcccagccctccgtctgtgtggtttg 181 aagcagaatttttccaccacatcctccactggacacccatcccaaatcagtctgaaagta 241 cctgctatgaagtggcgctcctgaggtatggaatagagtcctggaactccatctccaact 301 gtagccagaccctgtcctatgaccttaccgcagtgaccttggacctgtaccacagcaatg 361 gctaccgggccagagtgcgggctgtggacggcagccggcactccaactggaccgtcacca 421 acacccgcttctctgtggatgaagtgactctgacagttggcagtgtgaacctagagatcc 481 acaatggcttcatcctcgggaagattcagctacccaggcccaagatggcccccgcaaatg 541 acacatatgaaagcatcttcagtcacttccgagagtatgagattgccattcgcaaggtgc 601 cgggaaacttcacgttcacacacaagaaagtaaaacatgaaaacttcagcctcctaacct 661 ctggagaagtgggagagttctgtgtccaggtgaaaccatctgtcgcttcccgaagtaaca 721 aggggatgtggtctaaagaggagtgcatctccctcaccaggcagtatttcaccgtgacca 781 acgtcatcatcttctttgcctttgtcctgctgctctccggagccctcgcctactgcctgg 841 ccctccagctgtatgtgcggcgccgaaagaagctacccagtgtcctgctcttcaagaagc 901 ccagccccttcatcttcatcagccagcgtccctccccagagacccaagacaccatccacc 961 cgcttgatgaggaggcctttttgaaggtgtccccagagctgaagaacttggacctgcacg 1021 gcagcacagacagtggctttggcagcaccaagccatccctgcagactgaagagccccagt 1081 tcctcctccctgaccctcacccccaggctgacagaacgctgggaaacagggagccccctg 1141 tgctgggggacagctgcagtagtggcagcagcaatagcacagacagcgggatctgcctgc 1201 aggagcccagcctgagccccagcacagggcccacctgggagcaacaggtggggagcaaca 1261 gcaggggccaggatgacagtggcattgacttagttcaaaactctgagggccgggctgggg 1321 acacacagggtggctcggccttgggccaccacagtcccccggagcctgaggtgcctgggg 1381 aagaagacccagctgctgtggcattccagggttacctgaggcagaccagatgtgctgaag 1441 agaaggcaaccaagacaggctgcctggaggaagaatcgcccttgacagatggccttggcc 1501 ccaaattcgggagatgcctggttgatgaggcaggcttgcatccaccagccctggccaagg 1561 gctatttgaaacaggatcctctagaaatgactctggcttcctcaggggccccaacgggac 1621 agtggaaccagcccactgaggaatggtcactcctggccttgagcagctgcagtgacctgg 1681 gaatatctgactggagctttgcccatgaccttgcccctctaggctgtgtggcagccccag 1741 gtggtctcctgggcagctttaactcagacctggtcaccctgcccctcatctctagcctgc 1801 agtcaagtgagtgactcgggctgagaggctgcttttgattttagccatgcctgctcctct 1861 gcctggaccaggaggagggcccctggggcagaagttaggcacgaggcagtctgggcactt 1921 ttctgcaagtccactggggctggccccagccaggccctgcagggctggtcagggtgtctg 1981 gggcaggaggaggccaactcactgaactagtgcagggtatgtgggtggcactgacctgtt 2041 ctgttgactggggccctgcagactctggcagagctgagaagggcagggaccttctccctc 2101 ctaggaactctttcctgtatcataaaggattatttgctcaggggaaccatggggctttct 2161 ggagttgtggtgaggccaccaggctgaagtcagctcagacccagacctccctgcttaggc 2221 cactcgagcatcagagcttccagcaggaggaagggctgtaggaatggaagcttcagggcc 2281 ttgctgctggggtcatttttaggggaaaaaggaggatatgatggtcacatggggaacctc 2341 ccctcatcgggcctctggggcaggaagcttgtcactggaagatcttaaggtatatatttt 2401 ctggacactcaaacacatcataatggattcactgaggggagacaaagggagccgagaccc 2461 tggatggggcttccagctcagaacccatccctctggtgggtacctctggcacccatctgc 2521 aaatatctccctctctccaacaaatggagtagcatccccctggggcacttgctgaggcca 2581 agccactcacatcctcactttgctgccccaccatcttgctgacaacttccagagaagcca 2641 tggttttttgtattggtcataactcagccctttgggcggcctctgggcttgggcaccagc 2701 tcatgccagccccagagggtcagggttggaggcctgtgcttgtgtttgctgctaatgtcc 2761 agctacagacccagaggataagccactgggcactgggctggggtccctgccttgttggtg 2821 ttcagctgtgtgattttggactagccacttgtcagagggcctcaatctcccatctgtgaa 2881 ataaggactccacctttaggggaccctccatgtttgctgggtattagccaagctggtcct 2941 gggagaatgcagatactgtccgtggactaccaagctggcttgtttcttatgccagaggct 3001 aacagatccaatgggagtccatggtgtcatgccaagacagtatcagacacagccccagaa 3061 gggggcattatgggccctgcctccccataggccatttggactctgccttcaaacaaaggc 3121 agttcagtccacaggcatggaagctgtgaggggacaggcctgtgcgtgccatccagagtc 3181 atctcagccctgcctttctctggagcattctgaaaacagatattctggcccagggaatcc 3241 agccatgacccccacccctctgccaaagtactcttaggtgccagtctggtaactgaactc 3301 cctctggaggcaggcttgagggaggattcctcagggttcccttgaaagctttatttattt 3361 attttgttcatttatttattggagaggcagcattgcacagtgaaagaattctggatatct 3421 caggagccccgaaattctagctctgactttgctgtttccagtggtatgaccttggagaag 3481 tcacttatcctcttggagcctcagtttcctcatctgcagaataatgactgacttgtctaa 3541 ttcgtagggatgtgaggttctgctgaggaaatgggtatgaatgtgccttgaacacaaagc 3601 tctgtcaataagtgatacatgttttttattccaataaattgtcaagaccacaggaaaaaa 3661 aaaaaaaaaaaa HumanIL-10R-1cDNA,transcriptvariant2 (SEQIDNO:152) 1 gtcagtcccagcccaagggtagctggaggcgcgcaggccggctccgctccggccccggac 61 gatgcggcgcgcccaggatgctgccgtgcctcgtagtgctgctggcggcgctcctcagcc 121 tccgtcttggctcagacgctcatggctcacctgttgtggaagtggaagaggctgaaattg 181 acaggaactgacggattgggaaggatagagaagtatgcgcaaggccaaacccccaacccg 241 caaacctcatcatccacccacttctagatgagccggacagagctgcccagccctccgtct 301 gtgtggtttgaagcagaatttttccaccacatcctccactggacacccatcccaaatcag 361 tctgaaagtacctgctatgaagtggcgctcctgaggtatggaatagagtcctggaactcc 421 atctccaactgtagccagaccctgtcctatgaccttaccgcagtgaccttggacctgtac 481 cacagcaatggctaccgggccagagtgcgggctgtggacggcagccggcactccaactgg 541 accgtcaccaacacccgcttctctgtggatgaagtgactctgacagttggcagtgtgaac 601 ctagagatccacaatggcttcatcctcgggaagattcagctacccaggcccaagatggcc 661 cccgcaaatgacacatatgaaagcatcttcagtcacttccgagagtatgagattgccatt 721 cgcaaggtgccgggaaacttcacgttcacacacaagaaagtaaaacatgaaaacttcagc 781 ctcctaacctctggagaagtgggagagttctgtgtccaggtgaaaccatctgtcgcttcc 841 cgaagtaacaaggggatgtggtctaaagaggagtgcatctccctcaccaggcagtatttc 901 accgtgaccaacgtcatcatcttctttgcctttgtcctgctgctctccggagccctcgcc 961 tactgcctggccctccagctgtatgtgcggcgccgaaagaagctacccagtgtcctgctc 1021 ttcaagaagcccagccccttcatcttcatcagccagcgtccctccccagagacccaagac 1081 accatccacccgcttgatgaggaggcctttttgaaggtgtccccagagctgaagaacttg 1141 gacctgcacggcagcacagacagtggctttggcagcaccaagccatccctgcagactgaa 1201 gagccccagttcctcctccctgaccctcacccccaggctgacagaacgctgggaaacagg 1261 gagccccctgtgctgggggacagctgcagtagtggcagcagcaatagcacagacagcggg 1321 atctgcctgcaggagcccagcctgagccccagcacagggcccacctgggagcaacaggtg 1381 gggagcaacagcaggggccaggatgacagtggcattgacttagttcaaaactctgagggc 1441 cgggctggggacacacagggtggctcggccttgggccaccacagtcccccggagcctgag 1501 gtgcctggggaagaagacccagctgctgtggcattccagggttacctgaggcagaccaga 1561 tgtgctgaagagaaggcaaccaagacaggctgcctggaggaagaatcgcccttgacagat 1621 ggccttggccccaaattcgggagatgcctggttgatgaggcaggcttgcatccaccagcc 1681 ctggccaagggctatttgaaacaggatcctctagaaatgactctggcttcctcaggggcc 1741 ccaacgggacagtggaaccagcccactgaggaatggtcactcctggccttgagcagctgc 1801 agtgacctgggaatatctgactggagctttgcccatgaccttgcccctctaggctgtgtg 1861 gcagccccaggtggtctcctgggcagctttaactcagacctggtcaccctgcccctcatc 1921 tctagcctgcagtcaagtgagtgactcgggctgagaggctgcttttgattttagccatgc 1981 ctgctcctctgcctggaccaggaggagggcccctggggcagaagttaggcacgaggcagt 2041 ctgggcacttttctgcaagtccactggggctggccccagccaggccctgcagggctggtc 2101 agggtgtctggggcaggaggaggccaactcactgaactagtgcagggtatgtgggtggca 2161 ctgacctgttctgttgactggggccctgcagactctggcagagctgagaagggcagggac 2221 cttctccctcctaggaactctttcctgtatcataaaggattatttgctcaggggaaccat 2281 ggggctttctggagttgtggtgaggccaccaggctgaagtcagctcagacccagacctcc 2341 ctgcttaggccactcgagcatcagagcttccagcaggaggaagggctgtaggaatggaag 2401 cttcagggccttgctgctggggtcatttttaggggaaaaaggaggatatgatggtcacat 2461 ggggaacctcccctcatcgggcctctggggcaggaagcttgtcactggaagatcttaagg 2521 tatatattttctggacactcaaacacatcataatggattcactgaggggagacaaaggga 2581 gccgagaccctggatggggcttccagctcagaacccatccctctggtgggtacctctggc 2641 acccatctgcaaatatctccctctctccaacaaatggagtagcatccccctggggcactt 2701 gctgaggccaagccactcacatcctcactttgctgccccaccatcttgctgacaacttcc 2761 agagaagccatggttttttgtattggtcataactcagccctttgggcggcctctgggctt 2821 gggcaccagctcatgccagccccagagggtcagggttggaggcctgtgcttgtgtttgct 2881 gctaatgtccagctacagacccagaggataagccactgggcactgggctggggtccctgc 2941 cttgttggtgttcagctgtgtgattttggactagccacttgtcagagggcctcaatctcc 3001 catctgtgaaataaggactccacctttaggggaccctccatgtttgctgggtattagcca 3061 agctggtcctgggagaatgcagatactgtccgtggactaccaagctggcttgtttcttat 3121 gccagaggctaacagatccaatgggagtccatggtgtcatgccaagacagtatcagacac 3181 agccccagaagggggcattatgggccctgcctccccataggccatttggactctgccttc 3241 aaacaaaggcagttcagtccacaggcatggaagctgtgaggggacaggcctgtgcgtgcc 3301 atccagagtcatctcagccctgcctttctctggagcattctgaaaacagatattctggcc 3361 cagggaatccagccatgacccccacccctctgccaaagtactcttaggtgccagtctggt 3421 aactgaactccctctggaggcaggcttgagggaggattcctcagggttcccttgaaagct 3481 ttatttatttattttgttcatttatttattggagaggcagcattgcacagtgaaagaatt 3541 ctggatatctcaggagccccgaaattctagctctgactttgctgtttccagtggtatgac 3601 cttggagaagtcacttatcctcttggagcctcagtttcctcatctgcagaataatgactg 3661 acttgtctaattcgtagggatgtgaggttctgctgaggaaatgggtatgaatgtgccttg 3721 aacacaaagctctgtcaataagtgatacatgttttttattccaataaattgtcaagacca 3781 caggaaaaaaaaaaaaaaaaaa PrecursorHumanIL-10R-2Protein(withsignalsequenceinbold) (SEQIDNO:153) 1 mawslgswlggcllvsalgmvpppenvrmnsvnfknilqwespafakgnltftaqylsyr 61 ifqdkcmnttltecdfsslskygdhtlrvraefadehsdwvnitfcpvddtiigppgmqv 121 evladslhmrflapkieneyetwtmknvynswtynvqywkngtdekfqitpqydfevlrn 181 lepwttycvqvrgflpdrnkagewsepvceqtthdetvpswmvavilmasvfmvclallg 241 cfallwcvykktkyafsprnslpqhlkeflghphhntllffsfplsdendvfdklsviae 301 dsesgkqnpgdscslgtppgqgpqs HumanIL-10R-2cDNA (SEQIDNO:154) 1 cccgcccatctccgctggttcccggaagccgccgcggacaagctctcccgggcgcgggcg 61 ggggtcgtgtgcttggaggaagccgcggaacccccagcgtccgtccatggcgtggagcct 121 tgggagctggctgggtggctgcctgctggtgtcagcattgggaatggtaccacctcccga 181 aaatgtcagaatgaattctgttaatttcaagaacattctacagtgggagtcacctgcttt 241 tgccaaagggaacctgactttcacagctcagtacctaagttataggatattccaagataa 301 atgcatgaatactaccttgacggaatgtgatttctcaagtctttccaagtatggtgacca 361 caccttgagagtcagggctgaatttgcagatgagcattcagactgggtaaacatcacctt 421 ctgtcctgtggatgacaccattattggaccccctggaatgcaagtagaagtacttgctga 481 ttctttacatatgcgtttcttagcccctaaaattgagaatgaatacgaaacttggactat 541 gaagaatgtgtataactcatggacttataatgtgcaatactggaaaaacggtactgatga 601 aaagtttcaaattactccccagtatgactttgaggtcctcagaaacctggagccatggac 661 aacttattgtgttcaagttcgagggtttcttcctgatcggaacaaagctggggaatggag 721 tgagcctgtctgtgagcaaacaacccatgacgaaacggtcccctcctggatggtggccgt 781 catcctcatggcctcggtcttcatggtctgcctggcactcctcggctgcttcgccttgct 841 gtggtgcgtttacaagaagacaaagtacgccttctcccctaggaattctcttccacagca 901 cctgaaagagtttttgggccatcctcatcataacacacttctgtttttctcctttccatt 961 gtcggatgagaatgatgtttttgacaagctaagtgtcattgcagaagactctgagagcgg 1021 caagcagaatcctggtgacagctgcagcctcgggaccccgcctgggcaggggccccaaag 1081 ctaggctctgagaaggaaacacactcggctgggcacagtgacgtactccatctcacatct 1141 gcctcagtgagggatcagggcagcaaacaagggccaagaccatctgagccagccccacat 1201 ctagaactcccagaccctggacttagccaccagagagctacattttaaaggctgtcttgg 1261 caaaaatactccatttgggaactcactgccttataaaggctttcatgatgttttcagaag 1321 ttggccactgagagtgtaattttcagccttttatatcactaaaataagatcatgttttaa 1381 ttgtgagaaacagggccgagcacagtggctcacgcctgtaataccagcaccttagaggtc 1441 gaggcaggcggatcacttgaggtcaggagttcaagaccagcctggccaatatggtgaaac 1501 ccagtctctactaaaaatacaaaaattagctaggcatgatggcgcatgcctataatccca 1561 gctactcgagtgcctgaggcaggagaattgcatgaacccgggaggaggaggaggaggttg 1621 cagtgagccgagatagcggcactgcactccagcctgggtgacaaagtgagactccatctc 1681 aaaaaaaaaaaaaaaaaaaattgtgagaaacagaaatacttaaaatgaggaataagaatg 1741 gagatgttacatctggtagatgtaacattctaccagattatggatggactgatctgaaaa 1801 tcgacctcaactcaagggtggtcagctcaatgctacacagagcacggacttttggattct 1861 ttgcagtactttgaatttatttttctacctatatatgttttatatgctgctggtgctcca 1921 ttaaagttttactctgtgttgcactatatgtgttcatgataaaaaa

Recombinant IL-10

[0733] In some examples, an IL-10 receptor agonist is a recombinant IL-10 protein. In some examples, a recombinant IL-10 protein has an amino acid sequence that is identical to a human IL-10 protein (e.g., SEQ ID NO: 140). Non-limiting commercial sources of recombinant human IL-10 protein are available from Peprotech (Rocky Hill, N.J.), Novus Biologicals (Littleton, Colo.), Stemcell Technologies (Cambridge, Mass.), Millipore Sigma (Billerica, Mass.), and R&D Systems (Minneapolis, Minn.). In some examples, a recombinant human IL-10 protein can be Tenovil (Schering Corporation).

[0734] In some examples, a recombinant IL-10 protein is a functional fragment of human IL-10 protein (e.g., SEQ ID NO: 140). In some examples, a functional fragment of human IL-10 is a fragment of a human IL-10 protein (e.g., SEQ ID NO: 140) that is able to specifically bind to and activate a human receptor of IL-10. For example, a functional fragment of human IL-10 protein can have one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty amino acids from the N- and/or C-terminus of SEQ ID NO: 140.

[0735] In some examples, a recombinant human IL-10 includes a sequence at least 80% identical (e.g., at least 82% identical, at least 84% identical, at least 86% identical, at least 88% identical, at least 90% identical, at least 92% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 98% identical, or at least 99% identical) to SEQ ID NO: 140, and is able to specifically bind to and activate a human receptor of IL-10. Mutation of amino acids that are not conserved between different mammalian species less likely to have a negative effect on the activity of a recombinant IL-10 protein.

[0736] In some embodiments, the IL-10 receptor agonist is rhuIL-10 (Tenovil) or a variant thereof. See, e.g., McHutchison et al., J Interferon Cytokine Res. 1:1265-1270, 1999; Rosenblum et al., Regul. Toxicol. Pharmacol. 35:56-71, 2002; Schreiber et al., Gastroenterology 119(6):1461-1472, 2000; Maini et al., Arthritis Rheum. 40(Suppl):224, 1997.

[0737] Exemplary methods of making a recombinant human IL-10 are described in Pajkrt et al., J. Immunol. 158: 3971-3977, 1997). Additional exemplary methods of making recombinant IL-10 are described herein and are known in the art.

[0738] In some embodiments, a recombinant IL-10 is a pegylated recombinant IL-10 (e.g., pegylated recombinant human IL-10) (e.g., a 5 kDa N-terminally PEGylated form of IL-10; AM0010) (Infante et al., ASCO Meeting Abstracts 33(15_suppl):3017, 2015; Chan et al., PLoS One 11(6):e0156229, 2016; Mumm et al., Cancer Cell 20(6):781-796, 2011; Teng et al., Cancer Cell 20(6):691-693, 2011; U.S. Pat. Nos. 8,691,205; 8,865,652; 9,259,478; and 9,364,517; and U.S. Patent Application Publication Nos. 2008/0081031; 2009/0214471; 2011/0250163; 2011/0091419; 2014/0227223; 2015/0079031; 2015/0086505; 2016/0193352; 2016/0367689; 2016/0375101; and 2016/0166647).

[0739] In some embodiments, a recombinant IL-10 is a stabilized isoform of a recombinant IL-10. In some embodiments, the stabilized isoform of a recombinant IL-10 is a viral IL-10 protein (e.g., a human cytomegalovirus IL10 (e.g., cmv-IL10, LA-cmv-IL-10 (e.g., Lin et al., Virus Res. 131(2):213-223, 2008; Jenkins et al., J. Virol. 78(3):1440-1447, 2004; Kotenko et al., Proc. Natl. Acad. Sci. U.S.A. 97(4):1695-1700, 2000; Jones et al., Proc. Natl. Acad. Sci. U.S.A. 99(14):9404-9409, 2002) or a latency-associated viral IL-10 protein (e.g., Poole et al., J. Virol. 88(24):13947-13955, 2014).

[0740] In some embodiments, the recombinant IL-10 is a mammalian IL-10 homolog (see, e.g., WO 00/073457). In some embodiments, a mammalian IL-10 homolog is BCRF1, an EBV homolog of human IL-10, also known as viral IL-10, or a variant thereof (Liu et al., J. Immunol. 158(2):604-613, 1997).

Fusion Proteins

[0741] In some embodiments, the IL-10 receptor agonist is a fusion protein. In some embodiments, the fusion protein comprises the amino acid sequence of an IL-10 protein (or a functional fragment thereof) and a fusion partner (e.g., an Fc region (e.g., human IgG Fc) or human serum albumin). In some embodiments the fusion partner can be an antibody or an antigen-binding antibody fragment (e.g., an scFv) that targets IL-10 receptor agonist to an inflamed tissue. In some embodiments, the antibody or antigen-binding fragment that is a fusion partner can bind specifically, or preferentially, to inflamed gastrointestinal cells by, e.g., CD69. In some embodiments, an IL-10 receptor agonist that is a fusion protein can be, e.g., F8-IL-10, such as Dekavil (Philogen).

[0742] In some embodiments, the fusion protein is a L19-IL-10 fusion protein, a HyHEL10-IL-10 fusion protein, or a variant thereof. See, e.g., Trachsel et al., Arthritis Res. Ther. 9(1):R9, 2007, and Walmsley et al., Arthritis Rheum. 39: 495-503, 1996.

IL-10 Peptide Mimetic

[0743] In some embodiments, the IL-10 receptor agonist is an IL-10 peptide mimetic. Anon-limiting example of an IL-10 peptide mimetic is IT 9302 or a variant thereof (Osman et al., Surgery 124(3):584-92, 1998; Lopez et al., Immunobiology 216(10):1117-1126, 2011). Additional examples of IL-10 peptide mimetics are described in DeWitt, Nature Biotech. 17:214, 1999, and Reineke et al., Nature Biotech. 17:271-275, 1999.

Antibodies and Antigen-Binding Fragments

[0744] In some embodiments, the IL-10 receptor agonist is an antibody or an antigen-binding antibody fragment that binds to and activates an IL-10 receptor (e.g., a human IL-10 receptor). In some embodiments, the antibody or antigen-binding antibody fragment that specifically binds to an epitope on IL-10R-1 protein (e.g., human IL-10R-1 protein). In some embodiments, the antibody or antigen-binding antibody fragment that specifically binds to an epitope on IL-10R-2 protein (e.g., a human IL-10R-2 protein). In some embodiments, the antibody or the antigen-binding antibody fragment that specifically binds to an epitope on IL-10R-1 and IL-10R-2 proteins (e.g., human IL-10R-1 and human IL-10R-2 proteins).

[0745] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv).sub.2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv.sub.2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab).sub.2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0746] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0747] In some embodiments, the IL-10 receptor agonist is an antibody (e.g., F8-IL10 (also known as DEKAVIL) or a variant thereof (see, e.g., Schwager et al., Arthritis Res. Ther. 11(5):R142, 2009; Franz et al., Int. J. Cardiol. 195:311-322, 2015; Galeazzi et al., Isr. Med. Assoc. J. 16(10):666, 2014).

[0748] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0749] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9 M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7M, about 0.510.sup.7M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0750] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0751] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M-IS-1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1(inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1(inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1(inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Cells Producing a Recombinant IL-10

[0752] In some embodiments, any of the devices or compositions described herein can include a recombinant cell (e.g., a recombinant mammalian cell) that secretes a recombinant IL-10 (e.g., any of the recombinant IL-10 proteins described herein). In some embodiments, any of the devices or compositions described herein can include a cell (e.g., a mammalian cell) that secretes IL-10 (e.g., human IL-10). In some embodiments, the mammalian cell can be a mammalian cell obtained from the subject, and after introduction of a nucleic acid encoding the recombinant IL-10 (e.g., any of the recombinant IL-10 proteins described herein) into the cell obtained from the subject, the cell is incorporated into any of the compositions or devices described herein.

[0753] A recombinant cell can be generated by introducing a vector including a nucleic acid sequence encoding a recombinant IL-10 protein (e.g., any of the recombinant IL-10 proteins described herein). In some embodiments, the vector or the nucleic acid sequence encoding a recombinant IL-10 protein is integrated into a chromosome of the recombinant mammalian cell. In some embodiments, the vector or the nucleic acid sequence encoding a recombinant IL-10 protein is not integrated into a chromosome of the recombinant mammalian cell.

[0754] A vector can be a viral vector. Non-limiting examples of viral vectors include adenovirus vectors, herpes virus vectors, baculovirus vectors, and retroviral vectors. An expression vector can also be a plasmid or a cosmid. Additional examples of vectors are known in the art.

[0755] A vector can include a promoter sequence operably linked to the nucleic acid sequence encoding a recombinant IL-10 protein (e.g., any of the recombinant IL-10 proteins described herein). Non-limiting examples of promoter sequences that can be operably linked to the sequence (e.g., cDNA) encoding a recombinant IL-10 protein (e.g., any of the recombinant IL-10 proteins described herein) include: Simian Virus 40 (SV40) early promoter, ribosomal protein 21 (rpS21) promoter, hamster -actin promoter, cytomegalovirus (CMV) promoter (e.g., CMV immediate early promoter (see, e.g., Teschendorf et al., Anticancer Res. 22:3325-3330, 2002), ubiquitin C (UBC) promoter, elongation factor 1- (EF1A) promoter, phosphoenolpyruvate carboxykinase (PCK) promoter, IE2 promoter/enhancer region from mouse CMV (see, e.g., Chatellard et al., Biotechnol. Bioeng. 96:106-117, 2007), and chicken j-actin promoter. Additional non-limiting examples of human gene promoters that can be used in any of the vectors described herein are described in the Mammalian Promoter Database (Wistar Institute website at mrpombdb.wister.upenn.edu). Additional examples of mammalian promoter sequences that can be used in the expression vectors are known in the art.

[0756] Non-limiting examples of methods that can be used to introduce a vector or a nucleic acid into a cell (e.g., a mammalian cell) include lipofection, transfection, electroporation, microinjection, calcium phosphate transfection, dendrimer-based transfection, cationic polymer transfection, cell squeezing, sonoporation, optical transfection, impalection, hydrodynamic delivery, magnetofection, viral transduction (e.g., adenoviral and lentiviral transduction), and nanoparticle transfection. These and other methods of introducing a vector or a nucleic acid into a cell are well known in the art.

[0757] In some examples, the recombinant mammalian cell can be a Chinese Hamster Ovary (CHO) cell, a B cell, a CD8.sup.+ T cell, a dendritic cell, a keratinocyte or an epithelial cell. See, e.g., Mosser et al., Immunol. Rev. 226:205-218, 2009; Fillatreau et al., Nat. Rev. Immunol. 8:391-397, 2008; Ryan et al., Crit. Rev. Immunol. 27:15-32, 2007; Moore et al., Annu. Rev. Immunol. 19:683-765, 2001. In some embodiments, the recombinant mammalian cell can be a mesenchymal stem cell (e.g., Gupte et al., Biomed. J. 40(1):49-54, 2017).

[0758] Nucleic Acids and Vectors the Encode an IL-10 Receptor Agonist

[0759] In some examples, an IL-10 receptor agonist can be a nucleic acid (e.g., a vector) that includes a sequence encoding an IL-10 receptor agonist (e.g., any of the IL-10 proteins described herein). In some embodiments, the nucleic acid includes a sequence encoding IL-10 (e.g., human IL-10). In some embodiments, the nucleic acid includes a sequence encoding a recombinant IL-10 (e.g., a recombinant human IL-10). In some examples, the sequence encoding an IL-10 receptor agonist can be SEQ ID NO: 141. In some embodiments, the sequence encoding an IL-10 receptor agonist can include a sequence that is at least 80% (e.g., at least 82%, at least 84%, at least 86%, at least 88%, at least 90%, at least 92%, at least 94%, at least 95%, at least 96%, at least 98%, or at least 99%) identical to SEQ ID NO: 141.

[0760] The nucleic acid can be, e.g., a vector. In some embodiments, a vector can be a viral vector (e.g., an adenovirus vector, a herpes virus vector, a baculovirus vector, or a retrovirus vector). A vector can also be, e.g., a plasmid or a cosmid. Additional examples of vectors are known in the art.

[0761] A vector can include a promoter sequence operably linked to the sequence encoding an IL-10 receptor agonist (e.g., any of the recombinant IL-10 proteins described herein). Non-limiting examples of promoter sequences that can be operably linked to the sequence encoding an IL-10 receptor agonist (e.g., any of the recombinant IL-10 proteins described herein) include: Simian Virus 40 (SV40) early promoter, ribosomal protein 21 (rpS21) promoter, hamster -actin promoter, cytomegalovirus (CMV) promoter (e.g., CMV immediate early promoter (see, e.g., Teschendorf et al., Anticancer Res. 22:3325-3330, 2002), ubiquitin C (UBC) promoter, elongation factor 1- (EF1A) promoter, phosphoenolpyruvate carboxykinase (PCK) promoter, IE2 promoter/enhancer region from mouse CMV (see, e.g., Chatellard et al., Biotechnol. Bioeng. 96:106-117, 2007), and chicken -actin promoter. Additional non-limiting examples of human gene promoters that can be used in any of the vectors described herein are described in the Mammalian Promoter Database (Wistar Institute website at mrpombdb.wister.upenn.edu). A promoter can be a constitutive promoter or an inducible promoter. Examples of constitutive promoters and inducible promoters are known in the art. Additional examples and features of mammalian promoter sequences that can be used in the expression vectors are known in the art.

[0762] A non-limiting example of a composition including a nucleic acid that encodes an IL-10 receptor agonist is XT-150 (Xalud Therapeutics).

Additional Examples of IL-10 Receptor Agonists

[0763] In some embodiments, the recombinant cell is a recombinant Gram-positive bacterial cell (e.g., a genetically modified Lactococcus lactis (LL-Thy12) (see, e.g., Steidler et al., Science 289:1352-1355, 2000; Braat et al., Clin. Gastroenterol. Heptal. 4:754-759, 2006). In some embodiments, the recombinant cell is a recombinant Gram-negative bacterial cell (e.g., a Shigellaflexneri cell) that secretes an IL-10 receptor agonist (e.g., a recombinant IL-10 protein) (Chamekh et al., J. Immunol. 180(6): 4292-4298, 2008).

[0764] In some embodiments, the IL-10 receptor agonist is a cell (e.g., a Clostridium butyricum cell) that induces IL-10 production and secretion by a different cell (e.g., a macrophage) (e.g., Hayashi et al., Cell Host Microbe 13:711-722, 2013). In some embodiments, the IL-10 receptor agonist is a recombinant bacterial cell (e.g., a Lactobacillus acidophilus cell) that is deficient in lipoteichoic acid and induces IL-10 production and secretion by a different cell (e.g., a dendritic cell) (e.g., Mohamadzadeh et al., Proc. Natl. Acad. Sci. U.S.A. 108(suppl 1):4623-4630, 2011; Konstantinov et al., Proc. Natl. Acad. Sci. U.S.A. 105(49):19474-9, 2008). In some embodiments, the IL-10 receptor agonist is a bacterial cell or a fragment of a bacterial cell that is maintained in the supernatant that induces IL-10 secretion in a different cell (e.g., an immune cell) (e.g., a Faecalibacterium prausnitzii cell or a Faecalibacterium prausnitzii supernatant) (see, e.g., Sokol et al., Proc. Natl. Acad. Sci. U.S.A. 105(43):16731-16736, 2008).

[0765] Additional examples of other IL-10 receptor agonists are described in, e.g., U.S. Pat. No. 6,936,586; WO 96/01318; WO 91/00349; WO 13/130913; each incorporated in its entirety herein.

Integrin Inhibitors

[0766] The term integrin inhibitor refers to an agent which decreases the expression of one or more integrins and/or decreases the binding of an integrin ligand to one or more integrins that play a role in the recruitment, extravasation, and/or activation of a leukocyte. In some embodiments, the integrin inhibitor specifically binds to at least a portion of a ligand binding site on a target integrin. In some embodiments, the integrin inhibitor specifically binds to a target integrin at the same site as an endogenous ligand. In some embodiments, the integrin inhibitor decreases the level of expression of the target integrin in a mammalian cell. In some embodiments, the integrin inhibitor specifically binds to an integrin ligand.

[0767] Non-limiting examples of integrins that can be targeted by any of the integrin inhibitors described herein include: 21 integrin, 11 integrin, 47 integrin, integrin 41 (VLA-4), E-selectin, ICAM-1, 51 integrin, 41 integrin, VLA-4, 21 integrin, 53 integrin, 55 integrin, IIb3 integrin, and MAdCAM-1. A non-limiting example of integrin inhibitor that can decrease the expression and/or activity of 47 integrin is FTY720. A non-limiting example of an integrin inhibitor that specifically targets MAdCAM is PF-547659 (Pfizer). Non-limiting examples of an integrin inhibitor that specifically targets 47 is AJM300 (Ajinomoto), etrolizumab (Genentech), and vedolizumab (Millenium/Takeda).

[0768] In some embodiments, the integrin inhibitor is an IIb3 integrin inhibitor. In some embodiments, the IIb3 integrin inhibitor is abciximab (ReoPro, c7E3; Kononczuk et al., Curr. Drug Targets 16(13):1429-1437, 2015; Jiang et al., Appl. Microbiol. Biotechnol. 98(1):105-114, 2014), eptifibatide (Integrilin; Scarborough et al., J. Biol. Chem. 268:1066-1073, 1993; Tcheng et al., Circulation 91:2151-2157, 1995) or tirofiban (Aggrastat; Hartman et al., J. Med. Chem. 35:4640-4642, 1992; Pierro et al., Eur. J. Ophthalmol. 26(4):e74-76, 2016; Guan et al., Eur. J. Pharmacol 761:144-152, 2015). In some embodiments, the integrin inhibitor is an L-selective integrin inhibitor. In some embodiments, the integrin inhibitor is a 2 integrin inhibitor.

[0769] In some embodiments, the integrin inhibitor is an 4 integrin (e.g., an 41 integrin (e.g., Very Late Antigen-4 (VLA-4), CD49d, or CD29)) inhibitor, an 47 integrin inhibitor. In some embodiments, the integrin inhibitor targets endothelial VCAM1, fibronectin, mucosal addressin cellular adhesion molecule-1 (MAdCAM-1), vitronectin, tenascin-C, osteopontin (OPN), nephronectin, agiostatin, tissue-type transglutaminase, factor XIII, Von Willebrand factor (VWF), an ADAM protein, an ICAM protein, collagen, e-cadherin, laminin, fibulin-5, or TGF. In some embodiments, the 4 integrin inhibitor is natalizumab (Tysabri; Targan et al., Gastroenterology 132(5):1672-1683, 2007; Sandborn et al., N. Engl. J. Med. 353(18):1912-1925, 2005; Nakamura et al., Intern. Med. 56(2):211-214, 2017; and Singh et al., J Pediatr. Gastroenterol. Nutr. 62(6):863-866, 2016). In some embodiments, the integrin inhibitor is an endogenous integrin inhibitor (e.g., SHARPIN (Rantala et al., Nat. Cell. Biol. 13(11):1315-1324, 2011).

[0770] In some embodiments, the integrin inhibitor is an v integrin (e.g., an 51 integrin, an 53 integrin, an 55 integrin inhibitor, and/or an 56 integrin) inhibitor.

[0771] In some embodiments, the integrin inhibitor is an 51 integrin inhibitor.

[0772] In some embodiments, an integrin inhibitor is an inhibitory nucleic acid, an antibody or antigen-binding fragment thereof, a fusion protein, an integrin antagonist, a cyclic peptide, a disintegrin, a peptidomimetic, or a small molecule. In some embodiments, the inhibitory nucleic acid is a small hairpin RNA, a small interfering RNA, an antisense, an aptamer, or a microRNA.

Inhibitory Nucleic Acids

[0773] As described herein, inhibitory nucleic acids specifically bind (e.g., hybridize) to a nucleic acid encoding an integrin or an integrin ligand to treat inflammatory diseases (e.g., chronic inflammation, irritable bowel syndrome (IBS), rheumatoid arthritis, ulcerative colitis, Crohn's Disease, or auto-inflammatory disease). In some embodiments, the inhibitory nucleic acid can be an antisense nucleic acid, a ribozyme, a small interfering RNA, a small hairpin RNA, or a microRNA. Examples of aspects of these different inhibitory nucleic acids are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of a target integrin or a target integrin ligand (e.g., any of the exemplary target integrins or any of the exemplary integrin ligands described herein) in a mammalian cell can be synthesized in vitro.

[0774] Inhibitory nucleic acids that can decrease the expression of target integrin mRNA or a target integrin ligand mRNA (e.g., any of the exemplary integrins described herein or any of the exemplary integrin ligands described herein) in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of target integrin mRNA or a target integrin ligand mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 155-181).

TABLE-US-00018 Integrin2(ITGA)(NCBIRef.:NM_002203.3) (SEQIDNO:155) 1 ttttccctgctctcaccgggcgggggagagaagccctctggacagcttctagagtgtgca 61 ggttctcgtatccctcggccaagggtatcctctgcaaacctctgcaaacccagcgcaact 121 acggtcccccggtcagacccaggatggggccagaacggacaggggccgcgccgctgccgc 181 tgctgctggtgttagcgctcagtcaaggcattttaaattgttgtttggcctacaatgttg 241 gtctcccagaagcaaaaatattttccggtccttcaagtgaacagtttggctatgcagtgc 301 agcagtttataaatccaaaaggcaactggttactggttggttcaccctggagtggctttc 361 ctgagaaccgaatgggagatgtgtataaatgtcctgttgacctatccactgccacatgtg 421 aaaaactaaatttgcaaacttcaacaagcattccaaatgttactgagatgaaaaccaaca 481 tgagcctcggcttgatcctcaccaggaacatgggaactggaggttttctcacatgtggtc 541 ctctgtgggcacagcaatgtgggaatcagtattacacaacgggtgtgtgttctgacatca 601 gtcctgattttcagctctcagccagcttctcacctgcaactcagccctgcccttccctca 661 tagatgttgtggttgtgtgtgatgaatcaaatagtatttatccttgggatgcagtaaaga 721 attttttggaaaaatttgtacaaggcctggatataggccccacaaagacacaggtggggt 781 taattcagtatgccaataatccaagagttgtgtttaacttgaacacatataaaaccaaag 841 aagaaatgattgtagcaacatcccagacatcccaatatggtggggacctcacaaacacat 901 tcggagcaattcaatatgcaagaaaatatgcttattcagcagcttctggtgggcgacgaa 961 gtgctacgaaagtaatggtagttgtaactgacggtgaatcacatgatggttcaatgttga 1021 aagctgtgattgatcaatgcaaccatgacaatatactgaggtttggcatagcagttcttg 1081 ggtacttaaacagaaacgcccttgatactaaaaatttaataaaagaaataaaagcaatcg 1141 ctagtattccaacagaaagatactttttcaatgtgtctgatgaagcagctctactagaaa 1201 aggctgggacattaggagaacaaattttcagcattgaaggtactgttcaaggaggagaca 1261 actttcagatggaaatgtcacaagtgggattcagtgcagattactcttctcaaaatgata 1321 ttctgatgctgggtgcagtgggagcttttggctggagtgggaccattgtccagaagacat 1381 ctcatggccatttgatctttcctaaacaagcctttgaccaaattctgcaggacagaaatc 1441 acagttcatatttaggttactctgtggctgcaatttctactggagaaagcactcactttg 1501 ttgctggtgctcctcgggcaaattataccggccagatagtgctatatagtgtgaatgaga 1561 atggcaatatcacggttattcaggctcaccgaggtgaccagattggctcctattttggta 1621 gtgtgctgtgttcagttgatgtggataaagacaccattacagacgtgctcttggtaggtg 1681 caccaatgtacatgagtgacctaaagaaagaggaaggaagagtctacctgtttactatca 1741 aagagggcattttgggtcagcaccaatttcttgaaggccccgagggcattgaaaacactc 1801 gatttggttcagcaattgcagctctttcagacatcaacatggatggctttaatgatgtga 1861 ttgttggttcaccactagaaaatcagaattctggagctgtatacatttacaatggtcatc 1921 agggcactatccgcacaaagtattcccagaaaatcttgggatccgatggagcctttagga 1981 gccatctccagtactttgggaggtccttggatggctatggagatttaaatggggattcca 2041 tcaccgatgtgtctattggtgcctttggacaagtggttcaactctggtcacaaagtattg 2101 ctgatgtagctatagaagcttcattcacaccagaaaaaatcactttggtcaacaagaatg 2161 ctcagataattctcaaactctgcttcagtgcaaagttcagacctactaagcaaaacaatc 2221 aagtggccattgtatataacatcacacttgatgcagatggattttcatccagagtaacct 2281 ccagggggttatttaaagaaaacaatgaaaggtgcctgcagaagaatatggtagtaaatc 2341 aagcacagagttgccccgagcacatcatttatatacaggagccctctgatgttgtcaact 2401 ctttggatttgcgtgtggacatcagtctggaaaaccctggcactagccctgcccttgaag 2461 cctattctgagactgccaaggtcttcagtattcctttccacaaagactgtggtgaggacg 2521 gactttgcatttctgatctagtcctagatgtccgacaaataccagctgctcaagaacaac 2581 cctttattgtcagcaaccaaaacaaaaggttaacattttcagtaacgctgaaaaataaaa 2641 gggaaagtgcatacaacactggaattgttgttgatttttcagaaaacttgttttttgcat 2701 cattctccctgccggttgatgggacagaagtaacatgccaggtggctgcatctcagaagt 2761 ctgttgcctgcgatgtaggctaccctgctttaaagagagaacaacaggtgacttttacta 2821 ttaactttgacttcaatcttcaaaaccttcagaatcaggcgtctctcagtttccaagcct 2881 taagtgaaagccaagaagaaaacaaggctgataatttggtcaacctcaaaattcctctcc 2941 tgtatgatgctgaaattcacttaacaagatctaccaacataaatttttatgaaatctctt 3001 cggatgggaatgttccttcaatcgtgcacagttttgaagatgttggtccaaaattcatct 3061 tctccctgaaggtaacaacaggaagtgttccagtaagcatggcaactgtaatcatccaca 3121 tccctcagtataccaaagaaaagaacccactgatgtacctaactggggtgcaaacagaca 3181 aggctggtgacatcagttgtaatgcagatatcaatccactgaaaataggacaaacatctt 3241 cttctgtatctttcaaaagtgaaaatttcaggcacaccaaagaattgaactgcagaactg 3301 cttcctgtagtaatgttacctgctggttgaaagacgttcacatgaaaggagaatactttg 3361 ttaatgtgactaccagaatttggaacgggactttcgcatcatcaacgttccagacagtac 3421 agctaacggcagctgcagaaatcaacacctataaccctgagatatatgtgattgaagata 3481 acactgttacgattcccctgatgataatgaaacctgatgagaaagccgaagtaccaacag 3541 gagttataataggaagtataattgctggaatccttttgctgttagctctggttgcaattt 3601 tatggaagctcggcttcttcaaaagaaaatatgaaaagatgaccaaaaatccagatgaga 3661 ttgatgagaccacagagctcagtagctgaaccagcagacctacctgcagtgggaaccggc 3721 agcatcccagccagggtttgctgtttgcgtgaatggatttctttttaaatcccatatttt 3781 ttttatcatgtcgtaggtaaactaacctggtattttaagagaaaactgcaggtcagtttg 3841 gaatgaagaaattgtggggggtgggggaggtgcggggggcaggtagggaaataataggga 3901 aaatacctattttatatgatgggggaaaaaaagtaatctttaaactggctggcccagagt 3961 ttacattctaatttgcattgtgtcagaaacatgaaatgcttccaagcatgacaactttta 4021 aagaaaaatatgatactctcagattttaagggggaaaactgttctctttaaaatatttgt 4081 ctttaaacagcaactacagaagtggaagtgcttgatatgtaagtacttccacttgtgtat 4141 attttaatgaatattgatgttaacaagaggggaaaacaaaacacaggttttttcaattta 4201 tgctgctcatccaaagttgccacagatgatacttccaagtgataattttatttataaact 4261 aggtaaaatttgttgttggttccttttagaccacggctgccccttccacaccccatcttg 4321 ctctaatgatcaaaacatgcttgaataactgagcttagagtatacctcctatatgtccat 4381 ttaagttaggagagggggcgatatagagaataaggcacaaaattttgtttaaaactcaga 4441 atataacatgtaaaatcccatctgctagaagcccatcctgtgccagaggaaggaaaagga 4501 ggaaatttcctttctcttttaggaggcacaacagttctcttctaggatttgtttggctga 4561 ctggcagtaacctagtgaatttctgaaagatgagtaatttctttggcaaccttcctcctc 4621 ccttactgaaccactctcccacctcctggtggtaccattattatagaagccctctacagc 4681 ctgactttctctccagcggtccaaagttatcccctcctttacccctcatccaaagttccc 4741 actccttcaggacagctgctgtgcattagatattaggggggaaagtcatctgtttaattt 4801 acacacttgcatgaattactgtatataaactccttaacttcagggagctattttcattta 4861 gtgctaaacaagtaagaaaaataagctcgagtgaatttctaaatgttggaatgttatggg 4921 atgtaaacaatgtaaagtaagacatctcaggatttcaccagaagttacagatgaggcact 4981 ggaagccaccaaattagcaggtgcaccttctgtggctgtcttgtttctgaagtacttaaa 5041 cttccacaagagtgaatttgacctaggcaagtttgttcaaaaggtagatcctgagatgat 5101 ttggtcagattgggataaggcccagcaatctgcattttaacaagcaccccagtcactagg 5161 atgcagatggaccacactttgagaaacaccacccatttctactttttgcaccttattttc 5221 tctgttcctgagcccccacattctctaggagaaacttagaggaaaagggcacagacacta 5281 catatctaaagctttggacaagtccttgacctctataaacttcagagtcctcattataaa 5341 atgggaagactgagctggagttcagcagtgatgcttttagttttaaaagtctatgatctg 5401 gacttcctataatacaaatacacaatcctccaagaatttgacttggaaaaaaatgtcaaa 5461 ggaaaacaggttatctgcccatgtgcatatggacaaccttgactaccctggcctggcccg 5521 tggtggcagtccagggctatctgtactgtttacagaattactttgtagttgacaacacaa 5581 aacaaacaaaaaaggcataaaatgccagcggtttatagaaaaaacagcatggtattctcc 5641 agttaggtatgccagagtccaattcttttaacagctgtgagaatttgctgcttcattcca 5701 acaaaattttatttaaaaaaaaaaaaaaaagactggagaaactagtcattagcttgataa 5761 agaatatttaacagctagtggtgctggtgtgtacctgaagctccagctacttgagagact 5821 gagacaggaagatcgcttgagcccaggagttcaagtccagcctaagcaacatagcaagac 5881 cctgtctcaaaaaaatgactatttaaaaagacaatgtggccaggcacggtggctcacacc 5941 tgtaatcccaacactttgggaggctgaggccggtggatcacgaggtcaggagtttgagac 6001 tagcctggccaacatggtgaaaccccatctctaataatataaaaattagctgggcgtagt 6061 agcaggtgcctgtaatcccagttactcgggaagctgaggcaggagaatcacttgaacccg 6121 ggaggcagaggtttcagtgagccgagatcgcgccactgcactccagcctgggtgacaggg 6181 caagactctgtctcaaacaaacaaacaaaaaaaaagttagtactgtatatgtaaatacta 6241 gcttttcaatgtgctatacaaacaattatagcacatccttccttttactctgtctcacct 6301 cctttaggtgagtacttccttaaataagtgctaaacatacatatacggaacttgaaagct 6361 ttggttagccttgccttaggtaatcagcctagtttacactgtttccagggagtagttgaa 6421 ttactataaaccattagccacttgtctctgcaccatttatcacaccaggacagggtctct 6481 caacctgggcgctactgtcatttggggccaggtgattcttccttgcaggggctgtcctgt 6541 accttgtaggacagcagccctgtcctagaaggtatgtttagcagcattcctggcctctag 6601 ctacccgatgccagagcatgctccccccgcagtcatgacaatcaaaaaatgtctccagac 6661 attgtcaaatgcctcctggggggcagtatttctcaagcacttttaagcaaaggtaagtat 6721 tcatacaagaaatttagggggaaaaaacattgtttaaataaaagctatgtgttcctattc 6781 aacaatatttttgctttaaaagtaagtagagggcataaaagatgtcatattcaaatttcc 6841 atttcataaatggtgtacagacaaggtctatagaatgtggtaaaaacttgactgcaacac 6901 aaggcttataaaatagtaagatagtaaaatagcttatgaagaaactacagagatttaaaa 6961 ttgtgcatgactcatttcagcagcaaaataagaactcctaactgaacagaaatttttcta 7021 cctagcaatgttattcttgtaaaatagttacctattaaaactgtgaagagtaaaactaaa 7081 gccaatttattatagtcacacaagtgattatactaaaaattattataaaggttataattt 7141 tataatgtatttacctgtcctgatatatagctataacccaatatatgaaaatctcaaaaa 7201 ttaagacatcatcatacagaaggcaggattccttaaactgagatccctgatccatcttta 7261 atatttcaatttgcacacataaaacaatgcccttttgtgtacattcaggcatacccattt 7321 taatcaatttgaaaggttaatttaaacctctagaggtgaatgagaaacatgggggaaaag 7381 tatgaaataggtgaaaatcttaactatttctttgaactctaaagactgaaactgtagcca 7441 ttatgtaaataaagtttcatatgtacctgtttattttggcagattaagtcaaaatatgaa 7501 tgtatatattgcataactatgttagaattgtatatattttaaagaaattgtcttggatat 7561 tttcctttatacataatagataagtcttttttcaaatgtggtgtttgatgtttttgatta 7621 aatgtgttttgcctctttccacaaaaactgtaaaaataaatgcatgtttgtacaaaaagt 7681 tgcagaattcatttgatttatgagaaacaaaaattaaattgtagtcaacagttagtagtt 7741 tttctcatatccaagtataacaaacagaaaagtttcattattgtaacccacttttttcat 7801 accacattattgaatattgttacaattgttttgaaaataaagccattttctttgggcttt 7861 tataagttaaaaaaaaaa IntegrinIIb(2b)(NCBIRef.:NM_000419.4;SEQIDNO:156) 1 gctctgcccgttgctcagcaagttacttggggttccagtttgataagaaaagacttcctg 61 tggaggaatctgaagggaaggaggaggagctggcccattcctgcctgggaggttgtggaa 121 gaaggaagatggccagagctttgtgtccactgcaagccctctggcttctggagtgggtgc 181 tgctgctcttgggaccttgtgctgcccctccagcctgggccttgaacctggacccagtgc 241 agctcaccttctatgcaggccccaatggcagccagtttggattttcactggacttccaca 301 aggacagccatgggagagtggccatcgtggtgggcgccccgcggaccctgggccccagcc 361 aggaggagacgggcggcgtgttcctgtgcccctggagggccgagggcggccagtgcccct 421 cgctgctctttgacctccgtgatgagacccgaaatgtaggctcccaaactttacaaacct 481 tcaaggcccgccaaggactgggggcgtcggtcgtcagctggagcgacgtcattgtggcct 541 gcgccccctggcagcactggaacgtcctagaaaagactgaggaggctgagaagacgcccg 601 taggtagctgctttttggctcagccagagagcggccgccgcgccgagtactccccctgtc 661 gcgggaacaccctgagccgcatttacgtggaaaatgattttagctgggacaagcgttact 721 gtgaagcgggcttcagctccgtggtcactcaggccggagagctggtgcttggggctcctg 781 gcggctattatttcttaggtctcctggcccaggctccagttgcggatattttctcgagtt 841 accgcccaggcatccttttgtggcacgtgtcctcccagagcctctcctttgactccagca 901 acccagagtacttcgacggctactgggggtactcggtggccgtgggcgagttcgacgggg 961 atctcaacactacagaatatgtcgtcggtgcccccacttggagctggaccctgggagcgg 1021 tggaaattttggattcctactaccagaggctgcatcggctgcgcggagagcagatggcgt 1081 cgtattttgggcattcagtggctgtcactgacgtcaacggggatgggaggcatgatctgc 1141 tggtgggcgctccactgtatatggagagccgggcagaccgaaaactggccgaagtggggc 1201 gtgtgtatttgttcctgcagccgcgaggcccccacgcgctgggtgcccccagcctcctgc 1261 tgactggcacacagctctatgggcgattcggctctgccatcgcacccctgggcgacctcg 1321 accgggatggctacaatgacattgcagtggctgccccctacgggggtcccagtggccggg 1381 gccaagtgctggtgttcctgggtcagagtgaggggctgaggtcacgtccctcccaggtcc 1441 tggacagccccttccccacaggctctgcctttggcttctcccttcgaggtgccgtagaca 1501 tcgatgacaacggatacccagacctgatcgtgggagcttacggggccaaccaggtggctg 1561 tgtacagagctcagccagtggtgaaggcctctgtccagctactggtgcaagattcactga 1621 atcctgctgtgaagagctgtgtcctacctcagaccaagacacccgtgagctgcttcaaca 1681 tccagatgtgtgttggagccactgggcacaacattcctcagaagctatccctaaatgccg 1741 agctgcagctggaccggcagaagccccgccagggccggcgggtgctgctgctgggctctc 1801 aacaggcaggcaccaccctgaacctggatctgggcggaaagcacagccccatctgccaca 1861 ccaccatggccttccttcgagatgaggcagacttccgggacaagctgagccccattgtgc 1921 tcagcctcaatgtgtccctaccgcccacggaggctggaatggcccctgctgtcgtgctgc 1981 atggagacacccatgtgcaggagcagacacgaatcgtcctggactgtggggaagatgacg 2041 tatgtgtgccccagcttcagctcactgccagcgtgacgggctccccgctcctagttgggg 2101 cagataatgtcctggagctgcagatggacgcagccaacgagggcgagggggcctatgaag 2161 cagagctggccgtgcacctgccccagggcgcccactacatgcgggccctaagcaatgtcg 2221 agggctttgagagactcatctgtaatcagaagaaggagaatgagaccagggtggtgctgt 2281 gtgagctgggcaaccccatgaagaagaacgcccagataggaatcgcgatgttggtgagcg 2341 tggggaatctggaagaggctggggagtctgtgtccttccagctgcagatacggagcaaga 2401 acagccagaatccaaacagcaagattgtgctgctggacgtgccggtccgggcagaggccc 2461 aagtggagctgcgagggaactcctttccagcctccctggtggtggcagcagaagaaggtg 2521 agagggagcagaacagcttggacagctggggacccaaagtggagcacacctatgagctcc 2581 acaacaatggccctgggactgtgaatggtcttcacctcagcatccaccttccgggacagt 2641 cccagccctccgacctgctctacatcctggatatacagccccaggggggccttcagtgct 2701 tcccacagcctcctgtcaaccctctcaaggtggactgggggctgcccatccccagcccct 2761 cccccattcacccggcccatcacaagcgggatcgcagacagatcttcctgccagagcccg 2821 agcagccctcgaggcttcaggatccagttctcgtaagctgcgactcggcgccctgtactg 2881 tggtgcagtgtgacctgcaggagatggcgcgcgggcagcgggccatggtcacggtgctgg 2941 ccttcctgtggctgcccagcctctaccagaggcctctggatcagtttgtgctgcagtcgc 3001 acgcatggttcaacgtgtcctccctcccctatgcggtgcccccgctcagcctgccccgag 3061 gggaagctcaggtgtggacacagctgctccgggccttggaggagagggccattccaatct 3121 ggtgggtgctggtgggtgtgctgggtggcctgctgctgctcaccatcctggtcctggcca 3181 tgtggaaggtcggcttcttcaagcggaaccggccacccctggaagaagatgatgaagagg 3241 gggagtgatggtgcagcctacactattctagcaggagggttgggcgtgctacctgcaccg 3301 ccccttctccaacaagttgcctccaagctttgggttggagctgttccattgggtcctctt 3361 ggtgtcgtttccctcccaacagagctgggctaccccccctcctgctgcctaataaagaga 3421 ctgagccctgaaaaaaaaaaaaaaaaaaa Integrin4(VLA-4)(NCBIRef.:NM_000885.5;SEQIDNO:157) 1 ataacgtctttgtcactaaaatgttccccaggggccttcggcgagtctttttgtttggtt 61 ttttgtttttaatctgtggctcttgataatttatctagtggttgcctacacctgaaaaac 121 aagacacagtgtttaactatcaacgaaagaactggacggctccccgccgcagtcccactc 181 cccgagtttgtggctggcatttgggccacgccgggctgggcggtcacagcgaggggcgcg 241 cagtttggggtcacacagctccgcttctaggccccaaccaccgttaaaaggggaagcccg 301 tgccccatcaggtccgctcttgctgagcccagagccatcccgcgctctgcgggctgggag 361 gcccgggccaggacgcgagtcctgcgcagccgaggttccccagcgccccctgcagccgcg 421 cgtaggcagagacggagcccggccctgcgcctccgcaccacgcccgggaccccacccagc 481 ggcccgtacccggagaagcagcgcgagcacccgaagctcccggctggcggcagaaaccgg 541 gagtggggccgggcgagtgcgcggcatcccaggccggcccgaacgctccgcccgcggtgg 601 gccgacttcccctcctcttccctctctccttcctttagcccgctggcgccggacacgctg 661 cgcctcatctcttggggcgttcttccccgttggccaaccgtcgcatcccgtgcaactttg 721 gggtagtggccgtttagtgttgaatgttccccaccgagagcgcatggcttgggaagcgag 781 gcgcgaacccggcccccgaagggccgccgtccgggagacggtgatgctgttgctgtgcct 841 gggggtcccgaccggccgcccctacaacgtggacactgagagcgcgctgctttaccaggg 901 cccccacaacacgctgttcggctactcggtcgtgctgcacagccacggggcgaaccgatg 961 gctcctagtgggtgcgcccactgccaactggctcgccaacgcttcagtgatcaatcccgg 1021 ggcgatttacagatgcaggatcggaaagaatcccggccagacgtgcgaacagctccagct 1081 gggtagccctaatggagaaccttgtggaaagacttgtttggaagagagagacaatcagtg 1141 gttgggggtcacactttccagacagccaggagaaaatggatccatcgtgacttgtgggca 1201 tagatggaaaaatatattttacataaagaatgaaaataagctccccactggtggttgcta 1261 tggagtgccccctgatttacgaacagaactgagtaaaagaatagctccgtgttatcaaga 1321 ttatgtgaaaaaatttggagaaaattttgcatcatgtcaagctggaatatccagttttta 1381 cacaaaggatttaattgtgatgggggccccaggatcatcttactggactggctctctttt 1441 tgtctacaatataactacaaataaatacaaggcttttttagacaaacaaaatcaagtaaa 1501 atttggaagttatttaggatattcagtcggagctggtcattttcggagccagcatactac 1561 cgaagtagtcggaggagctcctcaacatgagcagattggtaaggcatatatattcagcat 1621 tgatgaaaaagaactaaatatcttacatgaaatgaaaggtaaaaagcttggatcgtactt 1681 tggagcttctgtctgtgctgtggacctcaatgcagatggcttctcagatctgctcgtggg 1741 agcacccatgcagagcaccatcagagaggaaggaagagtgtttgtgtacatcaactctgg 1801 ctcgggagcagtaatgaatgcaatggaaacaaacctcgttggaagtgacaaatatgctgc 1861 aagatttggggaatctatagttaatcttggcgacattgacaatgatggctttgaagatgt 1921 tgctatcggagctccacaagaagatgacttgcaaggtgctatttatatttacaatggccg 1981 tgcagatgggatctcgtcaaccttctcacagagaattgaaggacttcagatcagcaaatc 2041 gttaagtatgtttggacagtctatatcaggacaaattgatgcagataataatggctatgt 2101 agatgtagcagttggtgcttttcggtctgattctgctgtcttgctaaggacaagacctgt 2161 agtaattgttgacgcttctttaagccaccctgagtcagtaaatagaacgaaatttgactg 2221 tgttgaaaatggatggccttctgtgtgcatagatctaacactttgtttctcatataaggg 2281 caaggaagttccaggttacattgttttgttttataacatgagtttggatgtgaacagaaa 2341 ggcagagtctccaccaagattctatttctcttctaatggaacttctgacgtgattacagg 2401 aagcatacaggtgtccagcagagaagctaactgtagaacacatcaagcatttatgcggaa 2461 agatgtgcgggacatcctcaccccaattcagattgaagctgcttaccaccttggtcctca 2521 tgtcatcagtaaacgaagtacagaggaattcccaccacttcagccaattcttcagcagaa 2581 gaaagaaaaagacataatgaaaaaaacaataaactttgcaaggttttgtgcccatgaaaa 2641 ttgttctgctgatttacaggtttctgcaaagattgggtttttgaagccccatgaaaataa 2701 aacatatcttgctgttgggagtatgaagacattgatgttgaatgtgtccttgtttaatgc 2761 tggagatgatgcatatgaaacgactctacatgtcaaactacccgtgggtctttatttcat 2821 taagattttagagctggaagagaagcaaataaactgtgaagtcacagataactctggcgt 2881 ggtacaacttgactgcagtattggctatatatatgtagatcatctctcaaggatagatat 2941 tagctttctcctggatgtgagctcactcagcagagcggaagaggacctcagtatcacagt 3001 gcatgctacctgtgaaaatgaagaggaaatggacaatctaaagcacagcagagtgactgt 3061 agcaatacctttaaaatatgaggttaagctgactgttcatgggtttgtaaacccaacttc 3121 atttgtgtatggatcaaatgatgaaaatgagcctgaaacgtgcatggtggagaaaatgaa 3181 cttaactttccatgttatcaacactggcaatagtatggctcccaatgttagtgtggaaat 3241 aatggtaccaaattcttttagcccccaaactgataagctgttcaacattttggatgtcca 3301 gactactactggagaatgccactttgaaaattatcaaagagtgtgtgcattagagcagca 3361 aaagagtgcaatgcagaccttgaaaggcatagtccggttcttgtccaagactgataagag 3421 gctattgtactgcataaaagctgatccacattgtttaaatttcttgtgtaattttgggaa 3481 aatggaaagtggaaaagaagccagtgttcatatccaactggaaggccggccatccatttt 3541 agaaatggatgagacttcagcactcaagtttgaaataagagcaacaggttttccagagcc 3601 aaatccaagagtaattgaactaaacaaggatgagaatgttgcgcatgttctactggaagg 3661 actacatcatcaaagacccaaacgttatttcaccatagtgattatttcaagtagcttgct 3721 acttggacttattgtacttctgttgatctcatatgttatgtggaaggctggcttctttaa 3781 aagacaatacaaatctatcctacaagaagaaaacagaagagacagttggagttatatcaa 3841 cagtaaaagcaatgatgattaaggacttctttcaaattgagagaatggaaaacagactca 3901 ggttgtagtaaagaaatttaaaagacactgtttacaagaaaaaatgaattttgtttggac 3961 ttcttttactcatgatcttgtgacatattatgtcttcatgcaaggggaaaatctcagcaa 4021 tgattactctttgagatagaagaactgcaaaggtaataatacagccaaagataatctctc 4081 agcttttaaatgggtagagaaacactaaagcattcaatttattcaagaaaagtaagccct 4141 tgaagatatcttgaaatgaaagtataactgagttaaattatactggagaagtcttagact 4201 tgaaatactacttaccatatgtgcttgcctcagtaaaatgaaccccactgggtgggcaga 4261 ggttcatttcaaatacatctttgatacttgttcaaaatatgttctttaaaaatataattt 4321 tttagagagctgttcccaaattttctaacgagtggaccattatcactttaaagcccttta 4381 tttataatacatttcctacgggctgtgttccaacaaccattttttttcagcagactatga 4441 atattatagtattataggccaaactggcaaacttcagactgaacatgtacactggtttga 4501 gcttagtgaaattacttctggataattatttttttataattatggatttcaccatctttc 4561 tttctgtatatatacatgtgtttttatgtaggtatatatttaccattcttcctatctatt 4621 cttcctataacacacctttatcaagcatacccaggagtaatcttcaaatcttttgttata 4681 ttctgaaacaaaagattgtgagtgttgcactttacctgatacacgctgatttagaaaata 4741 cagaaaccatacctcactaataactttaaaatcaaagctgtgcaaagactagggggccta 4801 tacttcatatgtattatgtactatgtaaaatattgactatcacacaactatttccttgga 4861 tgtaattctttgttaccctttacaagtataagtgttaccttacatggaaacgaagaaaca 4921 aaattcataaatttaaattcataaatttagctgaaagatactgattcaatttgtatacag 4981 tgaatataaatgagacgacagcaaaattttcatgaaatgtaaaatatttttatagtttgt 5041 tcatactatatgaggttctattttaaatgactttctggattttaaaaaatttctttaaat 5101 acaatcatttttgtaatatttattttatgcttatgatctagataattgcagaatatcatt 5161 ttatctgactctgccttcataagagagctgtggccgaattttgaacatctgttataggga 5221 gtgatcaaattagaaggcaatgtggaaaaacaattctgggaaagatttctttatatgaag 5281 tccctgccactagccagccatcctaattgatgaaagttatctgttcacaggcctgcagtg 5341 atggtgaggaatgttctgagatttgcgaaggcatttgagtagtgaaatgtaagcacaaaa 5401 cctcctgaacccagagtgtgtatacacaggaataaactttatgacatttatgtattttta 5461 aaaaactttgtatcgttataaaaaggctagtcattctttcaggagaacatctaggatcat 5521 agatgaaaaatcaagccccgatttagaactgtcttctccaggatggtctctaaggaaatt 5581 tacatttggttctttcctactcagaactactcagaaacaactatatatttcaggttatct 5641 gagcacagtgaaagcagagtactatggttgtccaacacaggcctctcagatacaagggga 5701 acacaattacatattgggctagattttgcccagttcaaaatagtatttgttatcaactta 5761 ctttgttacttgtatcatgaattttaaaaccctaccactttaagaagacagggatgggtt 5821 attcttttttggcaggtaggctatataactatgtgattttgaaatttaactgctctggat 5881 tagggagcagtgaatcaaggcagacttatgaaatctgtattatatttgtaacagaatata 5941 ggaaatttaacataattgatgagctcaaatcctgaaaaatgaaagaatccaaattatttc 6001 agaattatctaggttaaatattgatgtattatgatggttgcaaagtttttttgtgtgtcc 6061 aataaacacattgtaaaaaaaagaatttgaattgatatctaaaaacagaatttgaattga 6121 tatttcatcttgacttttaaagccctagaggctaattgttagtaacatcaatttctatta 6181 ggatatccgtttggccacacagcaggaggttagagcaatggagcattactgagttcctcc 6241 ccctgtcagatcagcagcagcattagattctcatagaagtgcgaaccatatggtgaactg 6301 gtatgtgagggatctagagtgccatgttcctcaagagaatctaatgcctgatgatctgag 6361 gtggaacagttcatcctgaaaccattcccccatccacggaaaaattgtcttccatgaaac 6421 tggtcccaaaaagggtggggaccacaggtttaaagcatggccacatttctttatattaaa 6481 attctagtttgtacatttcttttagaaacaattacatgttactttggaatcatttcttcc 6541 atgcttcctccataaagactgataagtcttggatgcaatctgtaaagaaaatacattatt 6601 tcatcaacttattttgttgtttttcacatacacctaataagtatggtacacaatgccaat 6661 gccaaatacaaattgataacaaacacagcattcccaacagagctgtaatctagaaaactg 6721 agaaggtctgattgataaatcatcaacaacaataattgctctaaaacctccttaactgac 6781 ttccttgattgtccaatgctctccattacctctgtaaaacagtcagttatgcctctagaa 6841 cacccatgtctagtgggcacccctgcatgcttcttctaaccactgagtgtcacaatgcct 6901 accaagaatgcgtttgcaggttcctaaacctgtttataccagttgctatgtaaaattgtt 6961 cccaagggaagttgaatgctctgtaaaggcctaataaaagcaaattactgaacaaaacat 7021 gttacagtaattatgagtgagaggaaactaagatggaaggataaaaatctaacactttac 7081 tattcagatggctccactaaaagatttaagatcttgatccatttttaaaaatccaaaatg 7141 gaagttgtagacattatctgtagtttatgcacaacaataaattagaaagccaatgtagac 7201 acgcataaccaaagaaaatgccttgggtctacataacagttgaataaatgtaaagttgct 7261 tttaaaaaaaaaaaaaaaaaa Integrin5(NCBIRef.:NM_002205.4;SEQIDNO:158) 1 attcgcctctgggaggtttaggaagcggctccgggtcggtggccccaggacagggaagag 61 cgggcgctatggggagccggacgccagagtcccctctccacgccgtgcagctgcgctggg 121 gcccccggcgccgacccccgctgctgccgctgctgttgctgctgctgccgccgccaccca 181 gggtcgggggcttcaacttagacgcggaggccccagcagtactctcggggcccccgggct 241 ccttcttcggattctcagtggagttttaccggccgggaacagacggggtcagtgtgctgg 301 tgggagcacccaaggctaataccagccagccaggagtgctgcagggtggtgctgtctacc 361 tctgtccttggggtgccagccccacacagtgcacccccattgaatttgacagcaaaggct 421 ctcggctcctggagtcctcactgtccagctcagagggagaggagcctgtggagtacaagt 481 ccttgcagtggttcggggcaacagttcgagcccatggctcctccatcttggcatgcgctc 541 cactgtacagctggcgcacagagaaggagccactgagcgaccccgtgggcacctgctacc 601 tctccacagataacttcacccgaattctggagtatgcaccctgccgctcagatttcagct 661 gggcagcaggacagggttactgccaaggaggcttcagtgccgagttcaccaagactggcc 721 gtgtggttttaggtggaccaggaagctatttctggcaaggccagatcctgtctgccactc 781 aggagcagattgcagaatcttattaccccgagtacctgatcaacctggttcaggggcagc 841 tgcagactcgccaggccagttccatctatgatgacagctacctaggatactctgtggctg 901 ttggtgaattcagtggtgatgacacagaagactttgttgctggtgtgcccaaagggaacc 961 tcacttacggctatgtcaccatccttaatggctcagacattcgatccctctacaacttct 1021 caggggaacagatggcctcctactttggctatgcagtggccgccacagacgtcaatgggg 1081 acgggctggatgacttgctggtgggggcacccctgctcatggatcggacccctgacgggc 1141 ggcctcaggaggtgggcagggtctacgtctacctgcagcacccagccggcatagagccca 1201 cgcccacccttaccctcactggccatgatgagtttggccgatttggcagctccttgaccc 1261 ccctgggggacctggaccaggatggctacaatgatgtggccatcggggctccctttggtg 1321 gggagacccagcagggagtagtgtttgtatttcctgggggcccaggagggctgggctcta 1381 agccttcccaggttctgcagcccctgtgggcagccagccacaccccagacttctttggct 1441 ctgcccttcgaggaggccgagacctggatggcaatggatatcctgatctgattgtggggt 1501 cctttggtgtggacaaggctgtggtatacaggggccgccccatcgtgtccgctagtgcct 1561 ccctcaccatcttccccgccatgttcaacccagaggagcggagctgcagcttagagggga 1621 accctgtggcctgcatcaaccttagcttctgcctcaatgcttctggaaaacacgttgctg 1681 actccattggtttcacagtggaacttcagctggactggcagaagcagaagggaggggtac 1741 ggcgggcactgttcctggcctccaggcaggcaaccctgacccagaccctgctcatccaga 1801 atggggctcgagaggattgcagagagatgaagatctacctcaggaacgagtcagaatttc 1861 gagacaaactctcgccgattcacatcgctctcaacttctccttggacccccaagccccag 1921 tggacagccacggcctcaggccagccctacattatcagagcaagagccggatagaggaca 1981 aggctcagatcttgctggactgtggagaagacaacatctgtgtgcctgacctgcagctgg 2041 aagtgtttggggagcagaaccatgtgtacctgggtgacaagaatgccctgaacctcactt 2101 tccatgcccagaatgtgggtgagggtggcgcctatgaggctgagcttcgggtcaccgccc 2161 ctccagaggctgagtactcaggactcgtcagacacccagggaacttctccagcctgagct 2221 gtgactactttgccgtgaaccagagccgcctgctggtgtgtgacctgggcaaccccatga 2281 aggcaggagccagtctgtggggtggccttcggtttacagtccctcatctccgggacacta 2341 agaaaaccatccagtttgacttccagatcctcagcaagaatctcaacaactcgcaaagcg 2401 acgtggtttcctttcggctctccgtggaggctcaggcccaggtcaccctgaacggtgtct 2461 ccaagcctgaggcagtgctattcccagtaagcgactggcatccccgagaccagcctcaga 2521 aggaggaggacctgggacctgctgtccaccatgtctatgagctcatcaaccaaggcccca 2581 gctccattagccagggtgtgctggaactcagctgtccccaggctctggaaggtcagcagc 2641 tcctatatgtgaccagagttacgggactcaactgcaccaccaatcaccccattaacccaa 2701 agggcctggagttggatcccgagggttccctgcaccaccagcaaaaacgggaagctccaa 2761 gccgcagctctgcttcctcgggacctcagatcctgaaatgcccggaggctgagtgtttca 2821 ggctgcgctgtgagctcgggcccctgcaccaacaagagagccaaagtctgcagttgcatt 2881 tccgagtctgggccaagactttcttgcagcgggagcaccagccatttagcctgcagtgtg 2941 aggctgtgtacaaagccctgaagatgccctaccgaatcctgcctcggcagctgccccaaa 3001 aagagcgtcaggtggccacagctgtgcaatggaccaaggcagaaggcagctatggcgtcc 3061 cactgtggatcatcatcctagccatcctgtttggcctcctgctcctaggtctactcatct 3121 acatcctctacaagcttggattcttcaaacgctccctcccatatggcaccgccatggaaa 3181 aagctcagctcaagcctccagccacctctgatgcctgagtcctcccaatttcagactccc 3241 attcctgaagaaccagtccccccaccctcattctactgaaaaggaggggtctgggtactt 3301 cttgaaggtgctgacggccagggagaagctcctctccccagcccagagacatacttgaag 3361 ggccagagccaggggggtgaggagctggggatccctcccccccatgcactgtgaaggacc 3421 cttgtttacacataccctcttcatggatgggggaactcagatccagggacagaggcccca 3481 gcctccctgaagcctttgcattttggagagtttcctgaaacaacttggaaagataactag 3541 gaaatccattcacagttctttgggccagacatgccacaaggacttcctgtccagctccaa 3601 cctgcaaagatctgtcctcagccttgccagagatccaaaagaagcccccagctaagaacc 3661 tggaacttggggagttaagacctggcagctctggacagccccaccctggtgggccaacaa 3721 agaacactaactatgcatggtgccccaggaccagctcaggacagatgccacacaaggata 3781 gatgctggcccagggcccagagcccagctccaaggggaatcagaactcaaatggggccag 3841 atccagcctggggtctggagttgatctggaacccagactcagacattggcacctaatcca 3901 ggcagatccaggactatatttgggcctgctccagacctgatcctggaggcccagttcacc 3961 ctgatttaggagaagccaggaatttcccaggaccctgaaggggccatgatggcaacagat 4021 ctggaacctcagcctggccagacacaggccctccctgttccccagagaaaggggagccca 4081 ctgtcctgggcctgcagaatttgggttctgcctgccagctgcactgatgctgcccctcat 4141 ctctctgcccaacccttccctcaccttggcaccagacacccaggacttatttaaactctg 4201 ttgcaagtgcaataaatctgacccagtgcccccactgaccagaactagaaaaaaaaaaaa 4261 aaaaaaa Integrin1(NCBIRef.:NM_002211.3;SEQIDNO:159) 1 atcagacgcgcagaggaggcggggccgcggctggtttcctgccggggggcggctctgggc 61 cgccgagtcccctcctcccgcccctgaggaggaggagccgccgccacccgccgcgcccga 121 cacccgggaggccccgccagcccgcgggagaggcccagcgggagtcgcggaacagcaggc 181 ccgagcccaccgcgccgggccccggacgccgcgcggaaaagatgaatttacaaccaattt 241 tctggattggactgatcagttcagtttgctgtgtgtttgctcaaacagatgaaaatagat 301 gtttaaaagcaaatgccaaatcatgtggagaatgtatacaagcagggccaaattgtgggt 361 ggtgcacaaattcaacatttttacaggaaggaatgcctacttctgcacgatgtgatgatt 421 tagaagccttaaaaaagaagggttgccctccagatgacatagaaaatcccagaggctcca 481 aagatataaagaaaaataaaaatgtaaccaaccgtagcaaaggaacagcagagaagctca 541 agccagaggatattactcagatccaaccacagcagttggttttgcgattaagatcagggg 601 agccacagacatttacattaaaattcaagagagctgaagactatcccattgacctctact 661 accttatggacctgtcttactcaatgaaagacgatttggagaatgtaaaaagtcttggaa 721 cagatctgatgaatgaaatgaggaggattacttcggacttcagaattggatttggctcat 781 ttgtggaaaagactgtgatgccttacattagcacaacaccagctaagctcaggaaccctt 841 gcacaagtgaacagaactgcaccagcccatttagctacaaaaatgtgctcagtcttacta 901 ataaaggagaagtatttaatgaacttgttggaaaacagcgcatatctggaaatttggatt 961 ctccagaaggtggtttcgatgccatcatgcaagttgcagtttgtggatcactgattggct 1021 ggaggaatgttacacggctgctggtgttttccacagatgccgggtttcactttgctggag 1081 atgggaaacttggtggcattgttttaccaaatgatggacaatgtcacctggaaaataata 1141 tgtacacaatgagccattattatgattatccttctattgctcaccttgtccagaaactga 1201 gtgaaaataatattcagacaatttttgcagttactgaagaatttcagcctgtttacaagg 1261 agctgaaaaacttgatccctaagtcagcagtaggaacattatctgcaaattctagcaatg 1321 taattcagttgatcattgatgcatacaattccctttcctcagaagtcattttggaaaacg 1381 gcaaattgtcagaaggcgtaacaataagttacaaatcttactgcaagaacggggtgaatg 1441 gaacaggggaaaatggaagaaaatgttccaatatttccattggagatgaggttcaatttg 1501 aaattagcataacttcaaataagtgtccaaaaaaggattctgacagctttaaaattaggc 1561 ctctgggctttacggaggaagtagaggttattcttcagtacatctgtgaatgtgaatgcc 1621 aaagcgaaggcatccctgaaagtcccaagtgtcatgaaggaaatgggacatttgagtgtg 1681 gcgcgtgcaggtgcaatgaagggcgtgttggtagacattgtgaatgcagcacagatgaag 1741 ttaacagtgaagacatggatgcttactgcaggaaagaaaacagttcagaaatctgcagta 1801 acaatggagagtgcgtctgcggacagtgtgtttgtaggaagagggataatacaaatgaaa 1861 tttattctggcaaattctgcgagtgtgataatttcaactgtgatagatccaatggcttaa 1921 tttgtggaggaaatggtgtttgcaagtgtcgtgtgtgtgagtgcaaccccaactacactg 1981 gcagtgcatgtgactgttctttggatactagtacttgtgaagccagcaacggacagatct 2041 gcaatggccggggcatctgcgagtgtggtgtctgtaagtgtacagatccgaagtttcaag 2101 ggcaaacgtgtgagatgtgtcagacctgccttggtgtctgtgctgagcataaagaatgtg 2161 ttcagtgcagagccttcaataaaggagaaaagaaagacacatgcacacaggaatgttcct 2221 attttaacattaccaaggtagaaagtcgggacaaattaccccagccggtccaacctgatc 2281 ctgtgtcccattgtaaggagaaggatgttgacgactgttggttctattttacgtattcag 2341 tgaatgggaacaacgaggtcatggttcatgttgtggagaatccagagtgtcccactggtc 2401 cagacatcattccaattgtagctggtgtggttgctggaattgttcttattggccttgcat 2461 tactgctgatatggaagcttttaatgataattcatgacagaagggagtttgctaaatttg 2521 aaaaggagaaaatgaatgccaaatgggacacgggtgaaaatcctatttataagagtgccg 2581 taacaactgtggtcaatccgaagtatgagggaaaatgagtactgcccgtgcaaatcccac 2641 aacactgaatgcaaagtagcaatttccatagtcacagttaggtagctttagggcaatatt 2701 gccatggttttactcatgtgcaggttttgaaaatgtacaatatgtataatttttaaaatg 2761 ttttattattttgaaaataatgttgtaattcatgccagggactgacaaaagacttgagac 2821 aggatggttactcttgtcagctaaggtcacattgtgcctttttgaccttttcttcctgga 2881 ctattgaaatcaagcttattggattaagtgatatttctatagcgattgaaagggcaatag 2941 ttaaagtaatgagcatgatgagagtttctgttaatcatgtattaaaactgatttttagct 3001 ttacaaatatgtcagtttgcagttatgcagaatccaaagtaaatgtcctgctagctagtt 3061 aaggattgttttaaatctgttattttgctatttgcctgttagacatgactgatgacatat 3121 ctgaaagacaagtatgttgagagttgctggtgtaaaatacgtttgaaatagttgatctac 3181 aaaggccatgggaaaaattcagagagttaggaaggaaaaaccaatagctttaaaacctgt 3241 gtgccattttaagagttacttaatgtttggtaacttttatgccttcactttacaaattca 3301 agccttagataaaagaaccgagcaattttctgctaaaaagtccttgatttagcactattt 3361 acatacaggccatactttacaaagtatttgctgaatggggaccttttgagttgaatttat 3421 tttattatttttattttgtttaatgtctggtgctttctgtcacctcttctaatcttttaa 3481 tgtatttgtttgcaattttggggtaagactttttttatgagtactttttctttgaagttt 3541 tagcggtcaatttgcctttttaatgaacatgtgaagttatactgtggctatgcaacagct 3601 ctcacctacgcgagtcttactttgagttagtgccataacagaccactgtatgtttacttc 3661 tcaccatttgagttgcccatcttgtttcacactagtcacattcttgttttaagtgccttt 3721 agttttaacagttcactttttacagtgctatttactgaagttatttattaaatatgccta 3781 aaatacttaaatcggatgtcttgactctgatgtattttatcaggttgtgtgcatgaaatt 3841 tttatagattaaagaagttgaggaaaagcaaaaaaaaaa Integrin3(NCBIRef.:NM_000212.2;SEQIDNO:160) 1 cgccgcgggaggcggacgagatgcgagcgcggccgcggccccggccgctctgggcgactg 61 tgctggcgctgggggcgctggcgggcgttggcgtaggagggcccaacatctgtaccacgc 121 gaggtgtgagctcctgccagcagtgcctggctgtgagccccatgtgtgcctggtgctctg 181 atgaggccctgcctctgggctcacctcgctgtgacctgaaggagaatctgctgaaggata 241 actgtgccccagaatccatcgagttcccagtgagtgaggcccgagtactagaggacaggc 301 ccctcagcgacaagggctctggagacagctcccaggtcactcaagtcagtccccagagga 361 ttgcactccggctccggccagatgattcgaagaatttctccatccaagtgcggcaggtgg 421 aggattaccctgtggacatctactacttgatggacctgtcttactccatgaaggatgatc 481 tgtggagcatccagaacctgggtaccaagctggccacccagatgcgaaagctcaccagta 541 acctgcggattggcttcggggcatttgtggacaagcctgtgtcaccatacatgtatatct 601 ccccaccagaggccctcgaaaacccctgctatgatatgaagaccacctgcttgcccatgt 661 ttggctacaaacacgtgctgacgctaactgaccaggtgacccgcttcaatgaggaagtga 721 agaagcagagtgtgtcacggaaccgagatgccccagagggtggctttgatgccatcatgc 781 aggctacagtctgtgatgaaaagattggctggaggaatgatgcatcccacttgctggtgt 841 ttaccactgatgccaagactcatatagcattggacggaaggctggcaggcattgtccagc 901 ctaatgacgggcagtgtcatgttggtagtgacaatcattactctgcctccactaccatgg 961 attatccctctttggggctgatgactgagaagctatcccagaaaaacatcaatttgatct 1021 ttgcagtgactgaaaatgtagtcaatctctatcagaactatagtgagctcatcccaggga 1081 ccacagttggggttctgtccatggattccagcaatgtcctccagctcattgttgatgctt 1141 atgggaaaatccgttctaaagtagagctggaagtgcgtgacctccctgaagagttgtctc 1201 tatccttcaatgccacctgcctcaacaatgaggtcatccctggcctcaagtcttgtatgg 1261 gactcaagattggagacacggtgagcttcagcattgaggccaaggtgcgaggctgtcccc 1321 aggagaaggagaagtcctttaccataaagcccgtgggcttcaaggacagcctgatcgtcc 1381 aggtcacctttgattgtgactgtgcctgccaggcccaagctgaacctaatagccatcgct 1441 gcaacaatggcaatgggacctttgagtgtggggtatgccgttgtgggcctggctggctgg 1501 gatcccagtgtgagtgctcagaggaggactatcgcccttcccagcaggacgaatgcagcc 1561 cccgggagggtcagcccgtctgcagccagcggggcgagtgcctctgtggtcaatgtgtct 1621 gccacagcagtgactttggcaagatcacgggcaagtactgcgagtgtgacgacttctcct 1681 gtgtccgctacaagggggagatgtgctcaggccatggccagtgcagctgtggggactgcc 1741 tgtgtgactccgactggaccggctactactgcaactgtaccacgcgtactgacacctgca 1801 tgtccagcaatgggctgctgtgcagcggccgcggcaagtgtgaatgtggcagctgtgtct 1861 gtatccagccgggctcctatggggacacctgtgagaagtgccccacctgcccagatgcct 1921 gcacctttaagaaagaatgtgtggagtgtaagaagtttgaccggggagccctacatgacg 1981 aaaatacctgcaaccgttactgccgtgacgagattgagtcagtgaaagagcttaaggaca 2041 ctggcaaggatgcagtgaattgtacctataagaatgaggatgactgtgtcgtcagattcc 2101 agtactatgaagattctagtggaaagtccatcctgtatgtggtagaagagccagagtgtc 2161 ccaagggccctgacatcctggtggtcctgctctcagtgatgggggccattctgctcattg 2221 gccttgccgccctgctcatctggaaactcctcatcaccatccacgaccgaaaagaattcg 2281 ctaaatttgaggaagaacgcgccagagcaaaatgggacacagccaacaacccactgtata 2341 aagaggccacgtctaccttcaccaatatcacgtaccggggcacttaatgataagcagtca 2401 tcctcagatcattatcagcctgtgccacgattgcaggagtccctgccatcatgtttacag 2461 aggacagtatttgtggggagggatttggggctcagagtggggtaggttgggagaatgtca 2521 gtatgtggaagtgtgggtctgtgtgtgtgtatgtgggggtctgtgtgtttatgtgtgtgt 2581 gttgtgtgtgggagtgtgtaatttaaaattgtgatgtgtcctgataagctgagctcctta 2641 gcctttgtcccagaatgcctcctgcagggattcttcctgcttagcttgagggtgactatg 2701 gagctgagcaggtgttcttcattacctcagtgagaagccagctttcctcatcaggccatt 2761 gtccctgaagagaagggcagggctgaggcctctcattccagaggaagggacaccaagcct 2821 tggctctaccctgagttcataaatttatggttctcaggcctgactctcagcagctatggt 2881 aggaactgctgggcttggcagcccgggtcatctgtacctctgcctcctttcccctccctc 2941 aggccgaaggaggagtcagggagagctgaactattagagctgcctgtgccttttgccatc 3001 ccctcaacccagctatggttctctcgcaagggaagtccttgcaagctaattctttgacct 3061 gttgggagtgaggatgtctgggccactcaggggtcattcatggcctgggggatgtaccag 3121 catctcccagttcataatcacaacccttcagatttgccttattggcagctctactctgga 3181 ggtttgtttagaagaagtgtgtcacccttaggccagcaccatctctttacctcctaattc 3241 cacaccctcactgctgtagacatttgctatgagctggggatgtctctcatgaccaaatgc 3301 ttttcctcaaagggagagagtgctattgtagagccagaggtctggccctatgcttccggc 3361 ctcctgtccctcatccatagcacctccacatacctggccctgtgccttggtgtgctgtat 3421 ccatccatggggctgattgtatttaccttctacctcttggctgccttgtgaaggaattat 3481 tcccatgagttggctgggaataagtgccaggatggaatgatgggtcagttgtatcagcac 3541 gtgtggcctgttcttctatgggttggacaacctcattttaactcagtctttaatctgaga 3601 ggccacagtgcaattttattttatttttctcatgatgaggttttcttaacttaaaagaac 3661 atgtatataaacatgcttgcattatatttgtaaatttatgtgatggcaaagaaggagagc 3721 ataggaaaccacacagacttgggcagggtacagacactcccacttggcatcattcacagc 3781 aagtcactggccagtggctggatctgtgaggggctctctcatgatagaaggctatgggga 3841 tagatgtgtggacacattggacctttcctgaggaagagggactgttcttttgtcccagaa 3901 aagcagtggctccattggtgttgacatacatccaacattaaaagccacccccaaatgccc 3961 aagaaaaaaagaaagacttatcaacatttgttccatgagcagaaaactggagctctggcc 4021 tcagtgttacagctaaataatctttaattaaggcaagtcactttcttcttcttaaagctg 4081 ttttctagtttgagaaatgatgggattttagcagccagtcttgaaggtctctttcagtat 4141 caacattctaagatgctgggacttactgtgtcatcaaatgtgcggttaagattctctggg 4201 atattgatactgtttgtgtttttagttgggagatctgagagacctggctttggcaagagc 4261 agatgtcattccatatcacctttctcaatgaaagtctcattctatcctctctccaaaccc 4321 gttttccaacatttgttaatagttacgtctctcctgatgtagcacttaagcttcatttag 4381 ttattatttctttcttcactttgcacacatttgcatccacatattagggaagaggaatcc 4441 ataagtagctgaaatatctattctgtattattgtgttaacattgagaataagccttggaa 4501 ttagatatggggcaatgactgagccctgtctcacccatggattactccttactgtaggga 4561 atggcagtatggtagagggataaatagggggcggggagggatagtcatggatccaagaag 4621 tccttagaaatagtggcagggaacaggtgtggaagctcatgcctgtaattataaccttca 4681 gctactaagacaggtgtggtggctcacgcctgtgattataatcttcagttactaagacag 4741 agtccatgagagtgttaatgggacattttctttagataagatgttttatatgaagaaact 4801 gtatcaaagggggaagaaaatgtatttaacaggtgaatcaaatcaggaatcttgtctgag 4861 ctactggaatgaagttcacaggtcttgaagacca Integrin5(NCBIRef.:NM_002213.4;SEQIDNO:161) 1 gccgccgagcggagccagcccctcccctacccggagcagcccgctggggccgtcccgagc 61 ggcgacacactaggagtcccggccggccagccagggcagccgcggtcccgggactcggcc 121 gtgagtgctgcgggacggatggtggcggcggggcgcgggccagcgcgggcgccgtgagcc 181 ggagctgcgcgcggggcatgcggctgcggcccccggccctcggcccccgcgctccggccc 241 cagccccggccgccggcccccgcggagtgcagcgaccgcgccgccgctgagggaggcgcc 301 ccaccatgccgcgggccccggcgccgctgtacgcctgcctcctggggctctgcgcgctcc 361 tgccccggctcgcaggtctcaacatatgcactagtggaagtgccacctcatgtgaagaat 421 gtctgctaatccacccaaaatgtgcctggtgctccaaagaggacttcggaagcccacggt 481 ccatcacctctcggtgtgatctgagggcaaaccttgtcaaaaatggctgtggaggtgaga 541 tagagagcccagccagcagcttccatgtcctgaggagcctgcccctcagcagcaagggtt 601 cgggctctgcaggctgggacgtcattcagatgacaccacaggagattgccgtgaacctcc 661 ggcccggtgacaagaccaccttccagctacaggttcgccaggtggaggactatcctgtgg 721 acctgtactacctgatggacctctccctgtccatgaaggatgacttggacaatatccgga 781 gcctgggcaccaaactcgcggaggagatgaggaagctcaccagcaacttccggttgggat 841 ttgggtcttttgttgataaggacatctctcctttctcctacacggcaccgaggtaccaga 901 ccaatccgtgcattggttacaagttgtttccaaattgcgtcccctcctttgggttccgcc 961 atctgctgcctctcacagacagagtggacagcttcaatgaggaagttcggaaacagaggg 1021 tgtcccggaaccgagatgcccctgaggggggctttgatgcagtactccaggcagccgtct 1081 gcaaggagaagattggctggcgaaaggatgcactgcatttgctggtgttcacaacagatg 1141 atgtgccccacatcgcattggatggaaaattgggaggcctggtgcagccacacgatggcc 1201 agtgccacctgaacgaggccaacgagtacactgcatccaaccagatggactatccatccc 1261 ttgccttgcttggagagaaattggcagagaacaacatcaacctcatctttgcagtgacaa 1321 aaaaccattatatgctgtacaagaattttacagccctgatacctggaacaacggtggaga 1381 ttttagatggagactccaaaaatattattcaactgattattaatgcatacaatagtatcc 1441 ggtctaaagtggagttgtcagtctgggatcagcctgaggatcttaatctcttctttactg 1501 ctacctgccaagatggggtatcctatcctggtcagaggaagtgtgagggtctgaagattg 1561 gggacacggcatcttttgaagtatcattggaggcccgaagctgtcccagcagacacacgg 1621 agcatgtgtttgccctgcggccggtgggattccgggacagcctggaggtgggggtcacct 1681 acaactgcacgtgcggctgcagcgtggggctggaacccaacagcgccaggtgcaacggga 1741 gcgggacctatgtctgcggcctgtgtgagtgcagccccggctacctgggcaccaggtgcg 1801 agtgccaggatggggagaaccagagcgtgtaccagaacctgtgccgggaggcagagggca 1861 agccactgtgcagcgggcgtggggactgcagctgcaaccagtgctcctgcttcgagagcg 1921 agtttggcaagatctatgggcctttctgtgagtgcgacaacttctcctgtgccaggaaca 1981 agggagtcctctgctcaggccatggcgagtgtcactgcggggaatgcaagtgccatgcag 2041 gttacatcggggacaactgtaactgctcgacagacatcagcacatgccggggcagagatg 2101 gccagatctgcagcgagcgtgggcactgtctctgtgggcagtgccaatgcacggagccgg 2161 gggcctttggggagatgtgtgagaagtgccccacctgcccggatgcatgcagcaccaaga 2221 gagattgcgtcgagtgcctgctgctccactctgggaaacctgacaaccagacctgccaca 2281 gcctatgcagggatgaggtgatcacatgggtggacaccatcgtgaaagatgaccaggagg 2341 ctgtgctatgtttctacaaaaccgccaaggactgcgtcatgatgttcacctatgtggagc 2401 tccccagtgggaagtccaacctgaccgtcctcagggagccagagtgtggaaacaccccca 2461 acgccatgaccatcctcctggctgtggtcggtagcatcctccttgttgggcttgcactcc 2521 tggctatctggaagctgcttgtcaccatccacgaccggagggagtttgcaaagtttcaga 2581 gcgagcgatccagggcccgctatgaaatggcttcaaatccattatacagaaagcctatct 2641 ccacgcacactgtggacttcaccttcaacaagttcaacaaatcctacaatggcactgtgg 2701 actgatgtttccttctccgaggggctggagcggggatctgatgaaaaggtcagactgaaa 2761 cgccttgcacggctgctcggcttgatcacagctccctaggtaggcaccacagagaagacc 2821 ttctagtgagcctgggccaggagcccacagtgcctgtacaggaaggtgcctggccatgtc 2881 acctggctgctaggccagagccatgccaggctgcgtccctccgagcttgggataaagcaa 2941 ggggaccttggcgctctcagctttccctgccacatccagcttgttgtcccaatgaaatac 3001 tgagatgctgggctgtctctcccttccaggaatgctgggcccccagcctggccagacaag 3061 aagactgtcaggaagggtcggagtctgtaaaaccagcatacagtttggcttttttcacat 3121 tgatcatttttatatgaaataaaaagatcctgcatttatggtgtagttctgagtcctgag 3181 acttttctgcgtgatggctatgccttgcacacaggtgttggtgatggggctgttgagatg 3241 cctgttgaaggtacatcgtttgcaaatgtcagtttcctctcctgtccgtgtttgtttagt 3301 acttttataatgaaaagaaacaagattgtttgggattggaagtaaagattaaaaccaaaa 3361 gaatttgtgtttgtctgatactctctgtgtgtttctttctttctgagcggacttaaaatg 3421 gtgcccccagtggggattgaagcggccgtgtacttcctcagggatgggacacaggctggt 3481 ctgatactccagactgcagcttgtcaagtaagcatgaggtgctcggggcagtgagggctg 3541 tgcaagggggaacactgagcagatacctttggccccttccagcttttactgacagagagt 3601 tccaggctagacaccataaaaaccaccccttgttctgaggggctgaggctggaaatagat 3661 tgtacagacaagcaagggttgagtggtggttcccacacgaagtcatctcttaatcatcat 3721 tagcaatagcagttcccttccaaggcctcccctcactcccgaaacacttacgtcccatgc 3781 aggcccaatgcaaaaaaacacatttgagcttttttcccgcagggccatgaagtcccctta 3841 agttcccatatctaagatggttgactgaccctctccccttatgtacagaagaggaaactg 3901 attctcagagaggggaagtggcttgcccgagtgtttgttaggaggttactgaatgacaaa 3961 ctgttcctaagaccccatctcatgctggccagagggccagcctcctcattcctgcttgct 4021 cttagaaaatctttcactgatcattttttgtcactggaataacttcaaggttattatgct 4081 ttcattccaaatggatctgtcctcagctctggacccaattccccttacttcattttggca 4141 aacactaagtcaaatagtgaaatgcctgtcactacatagaacctattacctggggcaaat 4201 acgaacagattgagtttccttcatcttgtgtaaatatgatgaaacagagacctggtaact 4261 tggtgacactgttaaaccctttttgggataaagccaaatgtaaatgaaaacattaaacag 4321 ataaattgtggtgttgagacttttctgaattgagaaaaataaatgtaattttggaagaaa 4381 aaaaaaaaaaaa Integrin7(NCBIRef.:NM_000889.2;SEQIDNO:162) 1 aaatcttccccaccctggggagtgtcacttcctcctctgccgtctcccagatcagtacac 61 aaaggctgctgctgccgccagaggaaggactgctctgcacgcacctatgtggaaactaaa 121 gcccagagagaaagtctgacttgccccacagccagtgagtgactgcagcagcaccagaat 181 ctggtctgtttcctgtttggctcttctaccactacggcttgggatctcgggcatggtggc 241 tttgccaatggtccttgttttgctgctggtcctgagcagaggtgagagtgaattggacgc 301 caagatcccatccacaggggatgccacagaatggcggaatcctcacctgtccatgctggg 361 gtcctgccagccagccccctcctgccagaagtgcatcctctcacaccccagctgtgcatg 421 gtgcaagcaactgaacttcaccgcgtcgggagaggcggaggcgcggcgctgcgcccgacg 481 agaggagctgctggctcgaggctgcccgctggaggagctggaggagccccgcggccagca 541 ggaggtgctgcaggaccagccgctcagccagggcgcccgcggagagggtgccacccagct 601 ggcgccgcagcgggtccgggtcacgctgcggcctggggagccccagcagctccaggtccg 661 cttccttcgtgctgagggatacccggtggacctgtactaccttatggacctgagctactc 721 catgaaggacgacctggaacgcgtgcgccagctcgggcacgctctgctggtccggctgca 781 ggaagtcacccattctgtgcgcattggttttggttcctttgtggacaaaacggtgctgcc 841 ctttgtgagcacagtaccctccaaactgcgccacccctgccccacccggctggagcgctg 901 ccagtcaccattcagctttcaccatgtgctgtccctgacgggggacgcacaagccttcga 961 gcgggaggtggggcgccagagtgtgtccggcaatctggactcgcctgaaggtggcttcga 1021 tgccattctgcaggctgcactctgccaggagcagattggctggagaaatgtgtcccggct 1081 gctggtgttcacttcagacgacacattccatacagctggggacgggaagttgggcggcat 1141 tttcatgcccagtgatgggcactgccacttggacagcaatggcctctacagtcgcagcac 1201 agagtttgactacccttctgtgggtcaggtagcccaggccctctctgcagcaaatatcca 1261 gcccatctttgctgtcaccagtgccgcactgcctgtctaccaggagctgagtaaactgat 1321 tcctaagtctgcagttggggagctgagtgaggactccagcaacgtggtacagctcatcat 1381 ggatgcttataatagcctgtcttccaccgtgacccttgaacactcttcactccctcctgg 1441 ggtccacatttcttacgaatcccagtgtgagggtcctgagaagagggagggtaaggctga 1501 ggatcgaggacagtgcaaccacgtccgaatcaaccagacggtgactttctgggtttctct 1561 ccaagccacccactgcctcccagagccccatctcctgaggctccgggcccttggcttctc 1621 agaggagctgattgtggagttgcacacgctgtgtgactgtaattgcagtgacacccagcc 1681 ccaggctccccactgcagtgatggccagggacacctacaatgtggtgtatgcagctgtgc 1741 ccctggccgcctaggtcggctctgtgagtgctctgtggcagagctgtcctccccagacct 1801 ggaatctgggtgccgggctcccaatggcacagggcccctgtgcagtggaaagggtcactg 1861 tcaatgtggacgctgcagctgcagtggacagagctctgggcatctgtgcgagtgtgacga 1921 tgccagctgtgagcgacatgagggcatcctctgcggaggctttggtcgctgccaatgtgg 1981 agtatgtcactgtcatgccaaccgcacgggcagagcatgcgaatgcagtggggacatgga 2041 cagttgcatcagtcccgagggagggctctgcagtgggcatggacgctgcaaatgcaaccg 2101 ctgccagtgcttggacggctactatggtgctctatgcgaccaatgcccaggctgcaagac 2161 accatgcgagagacaccgggactgtgcagagtgtggggccttcaggactggcccactggc 2221 caccaactgcagtacagcttgtgcccataccaatgtgaccctggccttggcccctatctt 2281 ggatgatggctggtgcaaagagcggaccctggacaaccagctgttcttcttcttggtgga 2341 ggatgacgccagaggcacggtcgtgctcagagtgagaccccaagaaaagggagcagacca 2401 cacgcaggccattgtgctgggctgcgtagggggcatcgtggcagtggggctggggctggt 2461 cctggcttaccggctctcggtggaaatctatgaccgccgggaatacagtcgctttgagaa 2521 ggagcagcaacaactcaactggaagcaggacagtaatcctctctacaaaagtgccatcac 2581 gaccaccatcaatcctcgctttcaagaggcagacagtcccactctctgaaggagggaggg 2641 acacttacccaaggctcttctccttggaggacagtgggaactggagggtgagaggaaggg 2701 tgggtctgtaagaccttggtaggggactaattcactggcgaggtgcggccaccaccctac 2761 ttcattttcagagtgacacccaagagggctgcttcccatgcctgcaaccttgcatccatc 2821 tgggctaccccacccaagtatacaataaagtcttacctcagaccacaaaaaaaaaaaa E-selectin(NCBIRef.:NM_000450.2;SEQIDNO:163) 1 agctgttcttggctgacttcacatcaaaactcctatactgacctgagacagaggcagcag 61 tgatacccacctgagagatcctgtgtttgaacaactgcttcccaaaacggaaagtatttc 121 aagcctaaacctttgggtgaaaagaactcttgaagtcatgattgcttcacagtttctctc 181 agctctcactttggtgcttctcattaaagagagtggagcctggtcttacaacacctccac 241 ggaagctatgacttatgatgaggccagtgcttattgtcagcaaaggtacacacacctggt 301 tgcaattcaaaacaaagaagagattgagtacctaaactccatattgagctattcaccaag 361 ttattactggattggaatcagaaaagtcaacaatgtgtgggtctgggtaggaacccagaa 421 acctctgacagaagaagccaagaactgggctccaggtgaacccaacaataggcaaaaaga 481 tgaggactgcgtggagatctacatcaagagagaaaaagatgtgggcatgtggaatgatga 541 gaggtgcagcaagaagaagcttgccctatgctacacagctgcctgtaccaatacatcctg 601 cagtggccacggtgaatgtgtagagaccatcaataattacacttgcaagtgtgaccctgg 661 cttcagtggactcaagtgtgagcaaattgtgaactgtacagccctggaatcccctgagca 721 tggaagcctggtttgcagtcacccactgggaaacttcagctacaattcttcctgctctat 781 cagctgtgataggggttacctgccaagcagcatggagaccatgcagtgtatgtcctctgg 841 agaatggagtgctcctattccagcctgcaatgtggttgagtgtgatgctgtgacaaatcc 901 agccaatgggttcgtggaatgtttccaaaaccctggaagcttcccatggaacacaacctg 961 tacatttgactgtgaagaaggatttgaactaatgggagcccagagccttcagtgtacctc 1021 atctgggaattgggacaacgagaagccaacgtgtaaagctgtgacatgcagggccgtccg 1081 ccagcctcagaatggctctgtgaggtgcagccattcccctgctggagagttcaccttcaa 1141 atcatcctgcaacttcacctgtgaggaaggcttcatgttgcagggaccagcccaggttga 1201 atgcaccactcaagggcagtggacacagcaaatcccagtttgtgaagctttccagtgcac 1261 agccttgtccaaccccgagcgaggctacatgaattgtcttcctagtgcttctggcagttt 1321 ccgttatgggtccagctgtgagttctcctgtgagcagggttttgtgttgaagggatccaa 1381 aaggctccaatgtggccccacaggggagtgggacaacgagaagcccacatgtgaagctgt 1441 gagatgcgatgctgtccaccagcccccgaagggtttggtgaggtgtgctcattcccctat 1501 tggagaattcacctacaagtcctcttgtgccttcagctgtgaggagggatttgaattaca 1561 tggatcaactcaacttgagtgcacatctcagggacaatggacagaagaggttccttcctg 1621 ccaagtggtaaaatgttcaagcctggcagttccgggaaagatcaacatgagctgcagtgg 1681 ggagcccgtgtttggcactgtgtgcaagttcgcctgtcctgaaggatggacgctcaatgg 1741 ctctgcagctcggacatgtggagccacaggacactggtctggcctgctacctacctgtga 1801 agctcccactgagtccaacattcccttggtagctggactttctgctgctggactctccct 1861 cctgacattagcaccatttctcctctggcttcggaaatgcttacggaaagcaaagaaatt 1921 tgttcctgccagcagctgccaaagccttgaatcagatggaagctaccaaaagccttctta 1981 catcctttaagttcaaaagaatcagaaacaggtgcatctggggaactagagggatacact 2041 gaagttaacagagacagataactctcctcgggtctctggcccttcttgcctactatgcca 2101 gatgcctttatggctgaaaccgcaacacccatcaccacttcaatagatcaaagtccagca 2161 ggcaaggacggccttcaactgaaaagactcagtgttccctttcctactctcaggatcaag 2221 aaagtgttggctaatgaagggaaaggatattttcttccaagcaaaggtgaagagaccaag 2281 actctgaaatctcagaattccttttctaactctcccttgctcgctgtaaaatcttggcac 2341 agaaacacaatattttgtggctttctttcttttgcccttcacagtgtttcgacagctgat 2401 tacacagttgctgtcataagaatgaataataattatccagagtttagaggaaaaaaatga 2461 ctaaaaatattataacttaaaaaaatgacagatgttgaatgcccacaggcaaatgcatgg 2521 agggttgttaatggtgcaaatcctactgaatgctctgtgcgagggttactatgcacaatt 2581 taatcactttcatccctatgggattcagtgcttcttaaagagttcttaaggattgtgata 2641 tttttacttgcattgaatatattataatcttccatacttcttcattcaatacaagtgtgg 2701 tagggacttaaaaaacttgtaaatgctgtcaactatgatatggtaaaagttacttattct 2761 agattaccccctcattgtttattaacaaattatgttacatctgttttaaatttatttcaa 2821 aaagggaaactattgtcccctagcaaggcatgatgttaaccagaataaagttctgagtgt 2881 ttttactacagttgttttttgaaaacatggtagaattggagagtaaaaactgaatggaag 2941 gtttgtatattgtcagatattttttcagaaatatgtggtttccacgatgaaaaacttcca 3001 tgaggccaaacgttttgaactaataaaagcataaatgcaaacacacaaaggtataatttt 3061 atgaatgtctttgttggaaaagaatacagaaagatggatgtgctttgcattcctacaaag 3121 atgtttgtcagatatgatatgtaaacataattcttgtatattatggaagattttaaattc 3181 acaatagaaactcaccatgtaaaagagtcatctggtagatttttaacgaatgaagatgtc 3241 taatagttattccctatttgttttcttctgtatgttagggtgctctggaagagaggaatg 3301 cctgtgtgagcaagcatttatgtttatttataagcagatttaacaattccaaaggaatct 3361 ccagttttcagttgatcactggcaatgaaaaattctcagtcagtaattgccaaagctgct 3421 ctagccttgaggagtgtgagaatcaaaactctcctacacttccattaacttagcatgtgt 3481 tgaaaaaaaagtttcagagaagttctggctgaacactggcaacaacaaagccaacagtca 3541 aaacagagatgtgataaggatcagaacagcagaggttcttttaaaggggcagaaaaactc 3601 tgggaaataagagagaacaactactgtgatcaggctatgtatggaatacagtgttatttt 3661 ctttgaaattgtttaagtgttgtaaatatttatgtaaactgcattagaaattagctgtgt 3721 gaaataccagtgtggtttgtgtttgagttttattgagaattttaaattataacttaaaat 3781 attttataatttttaaagtatatatttatttaagcttatgtcagacctatttgacataac 3841 actataaaggttgacaataaatgtgcttatgttta ICAM-1(NCBIRef.:NM_000201.2;SEQIDNO:164) 1 caagcttagcctggccgggaaacgggaggcgtggaggccgggagcagcccccggggtcat 61 cgccctgccaccgccgcccgattgctttagcttggaaattccggagctgaagcggccagc 121 gagggaggatgaccctctcggcccgggcaccctgtcagtccggaaataactgcagcattt 181 gttccggaggggaaggcgcgaggtttccgggaaagcagcaccgccccttggcccccaggt 241 ggctagcgctataaaggatcacgcgccccagtcgacgctgagctcctctgctactcagag 301 ttgcaacctcagcctcgctatggctcccagcagcccccggcccgcgctgcccgcactcct 361 ggtcctgctcggggctctgttcccaggacctggcaatgcccagacatctgtgtccccctc 421 aaaagtcatcctgccccggggaggctccgtgctggtgacatgcagcacctcctgtgacca 481 gcccaagttgttgggcatagagaccccgttgcctaaaaaggagttgctcctgcctgggaa 541 caaccggaaggtgtatgaactgagcaatgtgcaagaagatagccaaccaatgtgctattc 601 aaactgccctgatgggcagtcaacagctaaaaccttcctcaccgtgtactggactccaga 661 acgggtggaactggcacccctcccctcttggcagccagtgggcaagaaccttaccctacg 721 ctgccaggtggagggtggggcaccccgggccaacctcaccgtggtgctgctccgtgggga 781 gaaggagctgaaacgggagccagctgtgggggagcccgctgaggtcacgaccacggtgct 841 ggtgaggagagatcaccatggagccaatttctcgtgccgcactgaactggacctgcggcc 901 ccaagggctggagctgtttgagaacacctcggccccctaccagctccagacctttgtcct 961 gccagcgactcccccacaacttgtcagcccccgggtcctagaggtggacacgcaggggac 1021 cgtggtctgttccctggacgggctgttcccagtctcggaggcccaggtccacctggcact 1081 gggggaccagaggttgaaccccacagtcacctatggcaacgactccttctcggccaaggc 1141 ctcagtcagtgtgaccgcagaggacgagggcacccagcggctgacgtgtgcagtaatact 1201 ggggaaccagagccaggagacactgcagacagtgaccatctacagctttccggcgcccaa 1261 cgtgattctgacgaagccagaggtctcagaagggaccgaggtgacagtgaagtgtgaggc 1321 ccaccctagagccaaggtgacgctgaatggggttccagcccagccactgggcccgagggc 1381 ccagctcctgctgaaggccaccccagaggacaacgggcgcagcttctcctgctctgcaac 1441 cctggaggtggccggccagcttatacacaagaaccagacccgggagcttcgtgtcctgta 1501 tggcccccgactggacgagagggattgtccgggaaactggacgtggccagaaaattccca 1561 gcagactccaatgtgccaggcttgggggaacccattgcccgagctcaagtgtctaaagga 1621 tggcactttcccactgcccatcggggaatcagtgactgtcactcgagatcttgagggcac 1681 ctacctctgtcgggccaggagcactcaaggggaggtcacccgcaaggtgaccgtgaatgt 1741 gctctccccccggtatgagattgtcatcatcactgtggtagcagccgcagtcataatggg 1801 cactgcaggcctcagcacgtacctctataaccgccagcggaagatcaagaaatacagact 1861 acaacaggcccaaaaagggacccccatgaaaccgaacacacaagccacgcctccctgaac 1921 ctatcccgggacagggcctcttcctcggccttcccatattggtggcagtggtgccacact 1981 gaacagagtggaagacatatgccatgcagctacacctaccggccctgggacgccggagga 2041 cagggcattgtcctcagtcagatacaacagcatttggggccatggtacctgcacacctaa 2101 aacactaggccacgcatctgatctgtagtcacatgactaagccaagaggaaggagcaaga 2161 ctcaagacatgattgatggatgttaaagtctagcctgatgagaggggaagtggtggggga 2221 gacatagccccaccatgaggacatacaactgggaaatactgaaacttgctgcctattggg 2281 tatgctgaggccccacagacttacagaagaagtggccctccatagacatgtgtagcatca 2341 aaacacaaaggcccacacttcctgacggatgccagcttgggcactgctgtctactgaccc 2401 caacccttgatgatatgtatttattcatttgttattttaccagctatttattgagtgtct 2461 tttatgtaggctaaatgaacataggtctctggcctcacggagctcccagtcctaatcaca 2521 ttcaaggtcaccaggtacagttgtacaggttgtacactgcaggagagtgcctggcaaaaa 2581 gatcaaatggggctgggacttctcattggccaacctgcctttccccagaaggagtgattt 2641 ttctatcggcacaaaagcactatatggactggtaatggttacaggttcagagattaccca 2701 gtgaggccttattcctcccttccccccaaaactgacacctttgttagccacctccccacc 2761 cacatacatttctgccagtgttcacaatgacactcagcggtcatgtctggacatgagtgc 2821 ccagggaatatgcccaagctatgccttgtcctcttgtcctgtttgcatttcactgggagc 2881 ttgcactatgcagctccagtttcctgcagtgatcagggtcctgcaagcagtggggaaggg 2941 ggccaaggtattggaggactccctcccagctttggaagcctcatccgcgtgtgtgtgtgt 3001 gtgtatgtgtagacaagctctcgctctgtcacccaggctggagtgcagtggtgcaatcat 3061 ggttcactgcagtcttgaccttttgggctcaagtgatcctcccacctcagcctcctgagt 3121 agctgggaccataggctcacaacaccacacctggcaaatttgattttttttttttttcca 3181 gagacggggtctcgcaacattgcccagacttcctttgtgttagttaataaagctttctca 3241 actgccaaa TGF-(NCBIRef.:NM_000660.6;SEQIDNO:165) 1 acctccctccgcggagcagccagacagcgagggccccggccgggggcaggggggacgccc 61 cgtccggggcacccccccggctctgagccgcccgcggggccggcctcggcccggagcgga 121 ggaaggagtcgccgaggagcagcctgaggccccagagtctgagacgagccgccgccgccc 181 ccgccactgcggggaggagggggaggaggagcgggaggagggacgagctggtcgggagaa 241 gaggaaaaaaacttttgagacttttccgttgccgctgggagccggaggcgcggggacctc 301 ttggcgcgacgctgccccgcgaggaggcaggacttggggaccccagaccgcctccctttg 361 ccgccggggacgcttgctccctccctgccccctacacggcgtccctcaggcgcccccatt 421 ccggaccagccctcgggagtcgccgacccggcctcccgcaaagacttttccccagacctc 481 gggcgcaccccctgcacgccgccttcatccccggcctgtctcctgagcccccgcgcatcc 541 tagaccctttctcctccaggagacggatctctctccgacctgccacagatcccctattca 601 agaccacccaccttctggtaccagatcgcgcccatctaggttatttccgtgggatactga 661 gacacccccggtccaagcctcccctccaccactgcgcccttctccctgaggacctcagct 721 ttccctcgaggccctcctaccttttgccgggagacccccagcccctgcaggggcggggcc 781 tccccaccacaccagccctgttcgcgctctcggcagtgccggggggcgccgcctccccca 841 tgccgccctccgggctgcggctgctgccgctgctgctaccgctgctgtggctactggtgc 901 tgacgcctggccggccggccgcgggactatccacctgcaagactatcgacatggagctgg 961 tgaagcggaagcgcatcgaggccatccgcggccagatcctgtccaagctgcggctcgcca 1021 gccccccgagccagggggaggtgccgcccggcccgctgcccgaggccgtgctcgccctgt 1081 acaacagcacccgcgaccgggtggccggggagagtgcagaaccggagcccgagcctgagg 1141 ccgactactacgccaaggaggtcacccgcgtgctaatggtggaaacccacaacgaaatct 1201 atgacaagttcaagcagagtacacacagcatatatatgttcttcaacacatcagagctcc 1261 gagaagcggtacctgaacccgtgttgctctcccgggcagagctgcgtctgctgaggctca 1321 agttaaaagtggagcagcacgtggagctgtaccagaaatacagcaacaattcctggcgat 1381 acctcagcaaccggctgctggcacccagcgactcgccagagtggttatcttttgatgtca 1441 ccggagttgtgcggcagtggttgagccgtggaggggaaattgagggctttcgccttagcg 1501 cccactgctcctgtgacagcagggataacacactgcaagtggacatcaacgggttcacta 1561 ccggccgccgaggtgacctggccaccattcatggcatgaaccggcctttcctgcttctca 1621 tggccaccccgctggagagggcccagcatctgcaaagctcccggcaccgccgagccctgg 1681 acaccaactattgcttcagctccacggagaagaactgctgcgtgcggcagctgtacattg 1741 acttccgcaaggacctcggctggaagtggatccacgagcccaagggctaccatgccaact 1801 tctgcctcgggccctgcccctacatttggagcctggacacgcagtacagcaaggtcctgg 1861 ccctgtacaaccagcataacccgggcgcctcggcggcgccgtgctgcgtgccgcaggcgc 1921 tggagccgctgcccatcgtgtactacgtgggccgcaagcccaaggtggagcagctgtcca 1981 acatgatcgtgcgctcctgcaagtgcagctgaggtcccgccccgccccgccccgccccgg 2041 caggcccggccccaccccgccccgcccccgctgccttgcccatgggggctgtatttaagg 2101 acacccgtgccccaagcccacctggggccccattaaagatggagagaggactgcggatct 2161 ctgtgtcattgggcgcctgcctggggtctccatccctgacgttcccccactcccactccc 2221 tctctctccctctctgcctcctcctgcctgtctgcactattcctttgcccggcatcaagg 2281 cacaggggaccagtggggaacactactgtagttagatctatttattgagcaccttgggca 2341 ctgttgaagtgccttacattaatgaactcattcagtcaccatagcaacactctgagatgc 2401 agggactctgataacacccattttaaaggtgaggaaacaagcccagagaggttaagggag 2461 gagttcctgcccaccaggaacctgctttagtgggggatagtgaagaagacaataaaagat 2521 agtagttcaggccaggcggggtggctcacgcctgtaatcctagcacttttgggaggcaga 2581 gatgggaggattacttgaatccaggcatttgagaccagcctgggtaacatagtgagaccc 2641 tatctctacaaaacacttttaaaaaatgtacacctgtggtcccagctactctggaggcta 2701 aggtgggaggatcacttgatcctgggaggtcaaggctgcag MadCAM-1(NCBIRef.:NM_130760.2;SEQIDNO:166) 1 gggactgagcatggatttcggactggccctcctgctggcggggcttctggggctcctcct 61 cggccagtccctccaggtgaagcccctgcaggtggagcccccggagccggtggtggccgt 121 ggccttgggcgcctcgcgccagctcacctgccgcctggcctgcgcggaccgcggggcctc 181 ggtgcagtggcggggcctggacaccagcctgggcgcggtgcagtcggacacgggccgcag 241 cgtcctcaccgtgcgcaacgcctcgctgtcggcggccgggacccgcgtgtgcgtgggctc 301 ctgcgggggccgcaccttccagcacaccgtgcagctccttgtgtacgccttcccggacca 361 gctgaccgtctccccagcagccctggtgcctggtgacccggaggtggcctgtacggccca 421 caaagtcacgcccgtggaccccaacgcgctctccttctccctgctcgtcgggggccagga 481 actggagggggcgcaagccctgggcccggaggtgcaggaggaggaggaggagccccaggg 541 ggacgaggacgtgctgttcagggtgacagagcgctggcggctgccgcccctggggacccc 601 tgtcccgcccgccctctactgccaggccacgatgaggctgcctggcttggagctcagcca 661 ccgccaggccatccccgtcctgcacagcccgacctccccggagcctcccgacaccacctc 721 cccggagtctcccgacaccacctccccggagtctcccgacaccacctcccaggagcctcc 781 cgacaccacctccccggagcctcccgacaagacctccccggagcccgccccccagcaggg 841 ctccacacacacccccaggagcccaggctccaccaggactcgccgccctgagatctccca 901 ggctgggcccacgcagggagaagtgatcccaacaggctcgtccaaacctgcgggtgacca 961 gctgcccgcggctctgtggaccagcagtgcggtgctgggactgctgctcctggccttgcc 1021 cacctatcacctctggaaacgctgccggcacctggctgaggacgacacccacccaccagc 1081 ttctctgaggcttctgccccaggtgtcggcctgggctgggttaagggggaccggccaggt 1141 cgggatcagcccctcctgagtggccagcctttccccctgtgaaagcaaaatagcttggac 1201 cccttcaagttgagaactggtcagggcaaacctgcctcccattctactcaaagtcatccc 1261 tctgttcacagagatggatgcatgttctgattgcctctttggagaagctcatcagaaact 1321 caaaagaaggccactgtttgtctcacctacccatgacctgaagcccctccctgagtggtc 1381 cccacctttctggacggaaccacgtactttttacatacattgattcatgtctcacgtctc 1441 cctaaaaatgcgtaagaccaagctgtgccctgaccaccctgggcccctgtcgtcaggacc 1501 tcctgaggctttggcaaataaacctcctaaaatgataaaaaaaaaa VCAM-1(NCBIRef.:NM_001078.3;SEQIDNO:167) 1 aaacttttttccctggctctgccctgggtttccccttgaagggatttccctccgcctctg 61 caacaagaccctttataaagcacagactttctatttcactccgcggtatctgcatcgggc 121 ctcactggcttcaggagctgaataccctcccaggcacacacaggtgggacacaaataagg 181 gttttggaaccactattttctcatcacgacagcaacttaaaatgcctgggaagatggtcg 241 tgatccttggagcctcaaatatactttggataatgtttgcagcttctcaagcttttaaaa 301 tcgagaccaccccagaatctagatatcttgctcagattggtgactccgtctcattgactt 361 gcagcaccacaggctgtgagtccccatttttctcttggagaacccagatagatagtccac 421 tgaatgggaaggtgacgaatgaggggaccacatctacgctgacaatgaatcctgttagtt 481 ttgggaacgaacactcttacctgtgcacagcaacttgtgaatctaggaaattggaaaaag 541 gaatccaggtggagatctactcttttcctaaggatccagagattcatttgagtggccctc 601 tggaggctgggaagccgatcacagtcaagtgttcagttgctgatgtatacccatttgaca 661 ggctggagatagacttactgaaaggagatcatctcatgaagagtcaggaatttctggagg 721 atgcagacaggaagtccctggaaaccaagagtttggaagtaacctttactcctgtcattg 781 aggatattggaaaagttcttgtttgccgagctaaattacacattgatgaaatggattctg 841 tgcccacagtaaggcaggctgtaaaagaattgcaagtctacatatcacccaagaatacag 901 ttatttctgtgaatccatccacaaagctgcaagaaggtggctctgtgaccatgacctgtt 961 ccagcgagggtctaccagctccagagattttctggagtaagaaattagataatgggaatc 1021 tacagcacctttctggaaatgcaactctcaccttaattgctatgaggatggaagattctg 1081 gaatttatgtgtgtgaaggagttaatttgattgggaaaaacagaaaagaggtggaattaa 1141 ttgttcaagagaaaccatttactgttgagatctcccctggaccccggattgctgctcaga 1201 ttggagactcagtcatgttgacatgtagtgtcatgggctgtgaatccccatctttctcct 1261 ggagaacccagatagacagccctctgagcgggaaggtgaggagtgaggggaccaattcca 1321 cgctgaccctgagccctgtgagttttgagaacgaacactcttatctgtgcacagtgactt 1381 gtggacataagaaactggaaaagggaatccaggtggagctctactcattccctagagatc 1441 cagaaatcgagatgagtggtggcctcgtgaatgggagctctgtcactgtaagctgcaagg 1501 ttcctagcgtgtacccccttgaccggctggagattgaattacttaagggggagactattc 1561 tggagaatatagagtttttggaggatacggatatgaaatctctagagaacaaaagtttgg 1621 aaatgaccttcatccctaccattgaagatactggaaaagctcttgtttgtcaggctaagt 1681 tacatattgatgacatggaattcgaacccaaacaaaggcagagtacgcaaacactttatg 1741 tcaatgttgcccccagagatacaaccgtcttggtcagcccttcctccatcctggaggaag 1801 gcagttctgtgaatatgacatgcttgagccagggctttcctgctccgaaaatcctgtgga 1861 gcaggcagctccctaacggggagctacagcctctttctgagaatgcaactctcaccttaa 1921 tttctacaaaaatggaagattctggggtttatttatgtgaaggaattaaccaggctggaa 1981 gaagcagaaaggaagtggaattaattatccaagttactccaaaagacataaaacttacag 2041 cttttccttctgagagtgtcaaagaaggagacactgtcatcatctcttgtacatgtggaa 2101 atgttccagaaacatggataatcctgaagaaaaaagcggagacaggagacacagtactaa 2161 aatctatagatggcgcctataccatccgaaaggcccagttgaaggatgcgggagtatatg 2221 aatgtgaatctaaaaacaaagttggctcacaattaagaagtttaacacttgatgttcaag 2281 gaagagaaaacaacaaagactatttttctcctgagcttctcgtgctctattttgcatcct 2341 ccttaataatacctgccattggaatgataatttactttgcaagaaaagccaacatgaagg 2401 ggtcatatagtcttgtagaagcacagaagtcaaaagtgtagctaatgcttgatatgttca 2461 actggagacactatttatctgtgcaaatccttgatactgctcatcattccttgagaaaaa 2521 caatgagctgagaggcagacttccctgaatgtattgaacttggaaagaaatgcccatcta 2581 tgtcccttgctgtgagcaagaagtcaaagtaaaacttgctgcctgaagaacagtaactgc 2641 catcaagatgagagaactggaggagttccttgatctgtatatacaataacataatttgta 2701 catatgtaaaataaaattatgccatagcaagattgcttaaaatagcaacactctatattt 2761 agattgttaaaataactagtgttgcttggactattataatttaatgcatgttaggaaaat 2821 ttcacattaatatttgctgacagctgacctttgtcatctttcttctattttattcccttt 2881 cacaaaattttattcctatatagtttattgacaataatttcaggttttgtaaagatgccg 2941 ggttttatatttttatagacaaataataagcaaagggagcactgggttgactttcaggta 3001 ctaaatacctcaacctatggtataatggttgactgggtttctctgtatagtactggcatg 3061 gtacggagatgtttcacgaagtttgttcatcagactcctgtgcaactttcccaatgtggc 3121 ctaaaaatgcaacttctttttattttcttttgtaaatgtttaggtttttttgtatagtaa 3181 agtgataatttctggaattagaaaaaaaaaaaaaaaaaaa Fibronectin(NCBIRef.:NM_001306129.1;SEQIDNO:168) 1 acgcccgcgccggctgtgctgcacagggggaggagagggaaccccaggcgcgagcgggaa 61 gaggggacctgcagccacaacttctctggtcctctgcatcccttctgtccctccacccgt 121 ccccttccccaccctctggcccccaccttcttggaggcgacaacccccgggaggcattag 181 aagggatttttcccgcaggttgcgaagggaagcaaacttggtggcaacttgcctcccggt 241 gcgggcgtctctcccccaccgtctcaacatgcttaggggtccggggcccgggctgctgct 301 gctggccgtccagtgcctggggacagcggtgccctccacgggagcctcgaagagcaagag 361 gcaggctcagcaaatggttcagccccagtccccggtggctgtcagtcaaagcaagcccgg 421 ttgttatgacaatggaaaacactatcagataaatcaacagtgggagcggacctacctagg 481 caatgcgttggtttgtacttgttatggaggaagccgaggttttaactgcgagagtaaacc 541 tgaagctgaagagacttgctttgacaagtacactgggaacacttaccgagtgggtgacac 601 ttatgagcgtcctaaagactccatgatctgggactgtacctgcatcggggctgggcgagg 661 gagaataagctgtaccatcgcaaaccgctgccatgaagggggtcagtcctacaagattgg 721 tgacacctggaggagaccacatgagactggtggttacatgttagagtgtgtgtgtcttgg 781 taatggaaaaggagaatggacctgcaagcccatagctgagaagtgttttgatcatgctgc 841 tgggacttcctatgtggtcggagaaacgtgggagaagccctaccaaggctggatgatggt 901 agattgtacttgcctgggagaaggcagcggacgcatcacttgcacttctagaaatagatg 961 caacgatcaggacacaaggacatcctatagaattggagacacctggagcaagaaggataa 1021 tcgaggaaacctgctccagtgcatctgcacaggcaacggccgaggagagtggaagtgtga 1081 gaggcacacctctgtgcagaccacatcgagcggatctggccccttcaccgatgttcgtgc 1141 agctgtttaccaaccgcagcctcacccccagcctcctccctatggccactgtgtcacaga 1201 cagtggtgtggtctactctgtggggatgcagtggctgaagacacaaggaaataagcaaat 1261 gctttgcacgtgcctgggcaacggagtcagctgccaagagacagctgtaacccagactta 1321 cggtggcaactcaaatggagagccatgtgtcttaccattcacctacaatggcaggacgtt 1381 ctactcctgcaccacagaagggcgacaggacggacatctttggtgcagcacaacttcgaa 1441 ttatgagcaggaccagaaatactctttctgcacagaccacactgttttggttcagactcg 1501 aggaggaaattccaatggtgccttgtgccacttccccttcctatacaacaaccacaatta 1561 cactgattgcacttctgagggcagaagagacaacatgaagtggtgtgggaccacacagaa 1621 ctatgatgccgaccagaagtttgggttctgccccatggctgcccacgaggaaatctgcac 1681 aaccaatgaaggggtcatgtaccgcattggagatcagtgggataagcagcatgacatggg 1741 tcacatgatgaggtgcacgtgtgttgggaatggtcgtggggaatggacatgcattgccta 1801 ctcgcagcttcgagatcagtgcattgttgatgacatcacttacaatgtgaacgacacatt 1861 ccacaagcgtcatgaagaggggcacatgctgaactgtacatgcttcggtcagggtcgggg 1921 caggtggaagtgtgatcccgtcgaccaatgccaggattcagagactgggacgttttatca 1981 aattggagattcatgggagaagtatgtgcatggtgtcagataccagtgctactgctatgg 2041 ccgtggcattggggagtggcattgccaacctttacagacctatccaagctcaagtggtcc 2101 tgtcgaagtatttatcactgagactccgagtcagcccaactcccaccccatccagtggaa 2161 tgcaccacagccatctcacatttccaagtacattctcaggtggagacctaaaaattctgt 2221 aggccgttggaaggaagctaccataccaggccacttaaactcctacaccatcaaaggcct 2281 gaagcctggtgtggtatacgagggccagctcatcagcatccagcagtacggccaccaaga 2341 agtgactcgctttgacttcaccaccaccagcaccagcacacctgtgaccagcaacaccgt 2401 gacaggagagacgactcccttttctcctcttgtggccacttctgaatctgtgaccgaaat 2461 cacagccagtagctttgtggtctcctgggtctcagcttccgacaccgtgtcgggattccg 2521 ggtggaatatgagctgagtgaggagggagatgagccacagtacctggatcttccaagcac 2581 agccacttctgtgaacatccctgacctgcttcctggccgaaaatacattgtaaatgtcta 2641 tcagatatctgaggatggggagcagagtttgatcctgtctacttcacaaacaacagcgcc 2701 tgatgcccctcctgacccgactgtggaccaagttgatgacacctcaattgttgttcgctg 2761 gagcagaccccaggctcccatcacagggtacagaatagtctattcgccatcagtagaagg 2821 tagcagcacagaactcaaccttcctgaaactgcaaactccgtcaccctcagtgacttgca 2881 acctggtgttcagtataacatcactatctatgctgtggaagaaaatcaagaaagtacacc 2941 tgttgtcattcaacaagaaaccactggcaccccacgctcagatacagtgccctctcccag 3001 ggacctgcagtttgtggaagtgacagacgtgaaggtcaccatcatgtggacaccgcctga 3061 gagtgcagtgaccggctaccgtgtggatgtgatccccgtcaacctgcctggcgagcacgg 3121 gcagaggctgcccatcagcaggaacacctttgcagaagtcaccgggctgtcccctggggt 3181 cacctattacttcaaagtctttgcagtgagccatgggagggagagcaagcctctgactgc 3241 tcaacagacaaccaaactggatgctcccactaacctccagtttgtcaatgaaactgattc 3301 tactgtcctggtgagatggactccacctcgggcccagataacaggataccgactgaccgt 3361 gggccttacccgaagaggacagcccaggcagtacaatgtgggtccctctgtctccaagta 3421 cccactgaggaatctgcagcctgcatctgagtacaccgtatccctcgtggccataaaggg 3481 caaccaagagagccccaaagccactggagtctttaccacactgcagcctgggagctctat 3541 tccaccttacaacaccgaggtgactgagaccaccattgtgatcacatggacgcctgctcc 3601 aagaattggttttaagctgggtgtacgaccaagccagggaggagaggcaccacgagaagt 3661 gacttcagactcaggaagcatcgttgtgtccggcttgactccaggagtagaatacgtcta 3721 caccatccaagtcctgagagatggacaggaaagagatgcgccaattgtaaacaaagtggt 3781 gacaccattgtctccaccaacaaacttgcatctggaggcaaaccctgacactggagtgct 3841 cacagtctcctgggagaggagcaccaccccagacattactggttatagaattaccacaac 3901 ccctacaaacggccagcagggaaattctttggaagaagtggtccatgctgatcagagctc 3961 ctgcacttttgataacctgagtcccggcctggagtacaatgtcagtgtttacactgtcaa 4021 ggatgacaaggaaagtgtccctatctctgataccatcatcccagaggtgccccaactcac 4081 tgacctaagctttgttgatataaccgattcaagcatcggcctgaggtggaccccgctaaa 4141 ctcttccaccattattgggtaccgcatcacagtagttgcggcaggagaaggtatccctat 4201 ttttgaagattttgtggactcctcagtaggatactacacagtcacagggctggagccggg 4261 cattgactatgatatcagcgttatcactctcattaatggcggcgagagtgcccctactac 4321 actgacacaacaaacggctgttcctcctcccactgacctgcgattcaccaacattggtcc 4381 agacaccatgcgtgtcacctgggctccacccccatccattgatttaaccaacttcctggt 4441 gcgttactcacctgtgaaaaatgaggaagatgttgcagagttgtcaatttctccttcaga 4501 caatgcagtggtcttaacaaatctcctgcctggtacagaatatgtagtgagtgtctccag 4561 tgtctacgaacaacatgagagcacacctcttagaggaagacagaaaacaggtcttgattc 4621 cccaactggcattgacttttctgatattactgccaactcttttactgtgcactggattgc 4681 tcctcgagccaccatcactggctacaggatccgccatcatcccgagcacttcagtgggag 4741 acctcgagaagatcgggtgccccactctcggaattccatcaccctcaccaacctcactcc 4801 aggcacagagtatgtggtcagcatcgttgctcttaatggcagagaggaaagtcccttatt 4861 gattggccaacaatcaacagtttctgatgttccgagggacctggaagttgttgctgcgac 4921 ccccaccagcctactgatcagctgggatgctcctgctgtcacagtgagatattacaggat 4981 cacttacggagagacaggaggaaatagccctgtccaggagttcactgtgcctgggagcaa 5041 gtctacagctaccatcagcggccttaaacctggagttgattataccatcactgtgtatgc 5101 tgtcactggccgtggagacagccccgcaagcagcaagccaatttccattaattaccgaac 5161 agaaattgacaaaccatcccagatgcaagtgaccgatgttcaggacaacagcattagtgt 5221 caagtggctgccttcaagttcccctgttactggttacagagtaaccaccactcccaaaaa 5281 tggaccaggaccaacaaaaactaaaactgcaggtccagatcaaacagaaatgactattga 5341 aggcttgcagcccacagtggagtatgtggttagtgtctatgctcagaatccaagcggaga 5401 gagtcagcctctggttcagactgcagtaaccaacattgatcgccctaaaggactggcatt 5461 cactgatgtggatgtcgattccatcaaaattgcttgggaaagcccacaggggcaagtttc 5521 caggtacagggtgacctactcgagccctgaggatggaatccatgagctattccctgcacc 5581 tgatggtgaagaagacactgcagagctgcaaggcctcagaccgggttctgagtacacagt 5641 cagtgtggttgccttgcacgatgatatggagagccagcccctgattggaacccagtccac 5701 agctattcctgcaccaactgacctgaagttcactcaggtcacacccacaagcctgagcgc 5761 ccagtggacaccacccaatgttcagctcactggatatcgagtgcgggtgacccccaagga 5821 gaagaccggaccaatgaaagaaatcaaccttgctcctgacagctcatccgtggttgtatc 5881 aggacttatggtggccaccaaatatgaagtgagtgtctatgctcttaaggacactttgac 5941 aagcagaccagctcagggagttgtcaccactctggagaatgtcagcccaccaagaagggc 6001 tcgtgtgacagatgctactgagaccaccatcaccattagctggagaaccaagactgagac 6061 gatcactggcttccaagttgatgccgttccagccaatggccagactccaatccagagaac 6121 catcaagccagatgtcagaagctacaccatcacaggtttacaaccaggcactgactacaa 6181 gatctacctgtacaccttgaatgacaatgctcggagctcccctgtggtcatcgacgcctc 6241 cactgccattgatgcaccatccaacctgcgtttcctggccaccacacccaattccttgct 6301 ggtatcatggcagccgccacgtgccaggattaccggctacatcatcaagtatgagaagcc 6361 tgggtctcctcccagagaagtggtccctcggccccgccctggtgtcacagaggctactat 6421 tactggcctggaaccgggaaccgaatatacaatttatgtcattgccctgaagaataatca 6481 gaagagcgagcccctgattggaaggaaaaagacagacgagcttccccaactggtaaccct 6541 tccacaccccaatcttcatggaccagagatcttggatgttccttccacagttcaaaagac 6601 ccctttcgtcacccaccctgggtatgacactggaaatggtattcagcttcctggcacttc 6661 tggtcagcaacccagtgttgggcaacaaatgatctttgaggaacatggttttaggcggac 6721 cacaccgcccacaacggccacccccataaggcataggccaagaccatacccgccgaatgt 6781 aggacaagaagctctctctcagacaaccatctcatgggccccattccaggacacttctga 6841 gtacatcatttcatgtcatcctgttggcactgatgaagaacccttacagttcagggttcc 6901 tggaacttctaccagtgccactctgacaggcctcaccagaggtgccacctacaacatcat 6961 agtggaggcactgaaagaccagcagaggcataaggttcgggaagaggttgttaccgtggg 7021 caactctgtcaacgaaggcttgaaccaacctacggatgactcgtgctttgacccctacac 7081 agtttcccattatgccgttggagatgagtgggaacgaatgtctgaatcaggctttaaact 7141 gttgtgccagtgcttaggctttggaagtggtcatttcagatgtgattcatctagatggtg 7201 ccatgacaatggtgtgaactacaagattggagagaagtgggaccgtcagggagaaaatgg 7261 ccagatgatgagctgcacatgtcttgggaacggaaaaggagaattcaagtgtgaccctca 7321 tgaggcaacgtgttatgatgatgggaagacataccacgtaggagaacagtggcagaagga 7381 atatctcggtgccatttgctcctgcacatgctttggaggccagcggggctggcgctgtga 7441 caactgccgcagacctgggggtgaacccagtcccgaaggcactactggccagtcctacaa 7501 ccagtattctcagagataccatcagagaacaaacactaatgttaattgcccaattgagtg 7561 cttcatgcctttagatgtacaggctgacagagaagattcccgagagtaaatcatctttcc 7621 aatccagaggaacaagcatgtctctctgccaagatccatctaaactggagtgatgttagc 7681 agacccagcttagagttcttctttctttcttaagccctttgctctggaggaagttctcca 7741 gcttcagctcaactcacagcttctccaagcatcaccctgggagtttcctgagggttttct 7801 cataaatgagggctgcacattgcctgttctgcttcgaagtattcaataccgctcagtatt 7861 ttaaatgaagtgattctaagatttggtttgggatcaataggaaagcatatgcagccaacc 7921 aagatgcaaatgttttgaaatgatatgaccaaaattttaagtaggaaagtcacccaaaca 7981 cttctgctttcacttaagtgtctggcccgcaatactgtaggaacaagcatgatcttgtta 8041 ctgtgatattttaaatatccacagtactcactttttccaaatgatcctagtaattgccta 8101 gaaatatctttctcttacctgttatttatcaatttttcccagtatttttatacggaaaaa 8161 attgtattgaaaacacttagtatgcagttgataagaggaatttggtataattatggtggg 8221 tgattattttttatactgtatgtgccaaagctttactactgtggaaagacaactgtttta 8281 ataaaagatttacattccacaacttgaagttcatctatttgatataagacaccttcgggg 8341 gaaataattcctgtgaatattctttttcaattcagcaaacatttgaaaatctatgatgtg 8401 caagtctaattgttgatttcagtacaagattttctaaatcagttgctacaaaaactgatt 8461 ggtttttgtcacttcatctcttcactaatggagatagctttacactttctgctttaatag 8521 atttaagtggaccccaatatttattaaaattgctagtttaccgttcagaagtataataga 8581 aataatctttagttgctcttttctaaccattgtaattcttcccttcttccctccaccttt 8641 ccttcattgaataaacctctgttcaaagagattgcctgcaagggaaataaaaatgactaa 8701 gatattaaaagtatttgaatagtaaaaaaaaaaaaaaaaaaa Vitronectin(NCBIRef.:NM_000638.3;SEQIDNO:169) 1 gagcaaacagagcagcagaaaaggcagttcctcttctccagtgccctccttccctgtctc 61 tgcctctccctcccttcctcaggcatcagagcggagacttcagggagaccagagcccagc 121 ttgccaggcactgagctagaagccctgccatggcacccctgagaccccttctcatactgg 181 ccctgctggcatgggttgctctggctgaccaagagtcatgcaagggccgctgcactgagg 241 gcttcaacgtggacaagaagtgccagtgtgacgagctctgctcttactaccagagctgct 301 gcacagactatacggctgagtgcaagccccaagtgactcgcggggatgtgttcactatgc 361 cggaggatgagtacacggtctatgacgatggcgaggagaaaaacaatgccactgtccatg 421 aacaggtggggggcccctccctgacctctgacctccaggcccagtccaaagggaatcctg 481 agcagacacctgttctgaaacctgaggaagaggcccctgcgcctgaggtgggcgcctcta 541 agcctgaggggatagactcaaggcctgagacccttcatccagggagacctcagcccccag 601 cagaggaggagctgtgcagtgggaagcccttcgacgccttcaccgacctcaagaacggtt 661 ccctctttgccttccgagggcagtactgctatgaactggacgaaaaggcagtgaggcctg 721 ggtaccccaagctcatccgagatgtctggggcatcgagggccccatcgatgccgccttca 781 cccgcatcaactgtcaggggaagacctacctcttcaagggtagtcagtactggcgctttg 841 aggatggtgtcctggaccctgattacccccgaaatatctctgacggcttcgatggcatcc 901 cggacaacgtggatgcagccttggccctccctgcccatagctacagtggccgggagcggg 961 tctacttcttcaaggggaaacagtactgggagtaccagttccagcaccagcccagtcagg 1021 aggagtgtgaaggcagctccctgtcggctgtgtttgaacactttgccatgatgcagcggg 1081 acagctgggaggacatcttcgagcttctcttctggggcagaacctctgctggtaccagac 1141 agccccagttcattagccgggactggcacggtgtgccagggcaagtggacgcagccatgg 1201 ctggccgcatctacatctcaggcatggcaccccgcccctccttggccaagaaacaaaggt 1261 ttaggcatcgcaaccgcaaaggctaccgttcacaacgaggccacagccgtggccgcaacc 1321 agaactcccgccggccatcccgcgccacgtggctgtccttgttctccagtgaggagagca 1381 acttgggagccaacaactatgatgactacaggatggactggcttgtgcctgccacctgtg 1441 aacccatccagagtgtcttcttcttctctggagacaagtactaccgagtcaatcttcgca 1501 cacggcgagtggacactgtggaccctccctacccacgctccatcgctcagtactggctgg 1561 gctgcccagctcctggccatctgtaggagtcagagcccacatggccgggccctctgtagc 1621 tccctcctcccatctccttcccccagcccaataaaggtcccttagccccgagtttaaa Tenascin-C(NCBIRef.:NM_002160.3;SEQIDNO:170) 1 aattcgccaactgaaaaagtgggaaaggatgtctggaggcgaggcgtcccattacagagg 61 aaggagctcgctatataagccagccaaagttggctgcaccggccacagcctgcctactgt 121 cacccgcctctcccgcgcgcagatacacgcccccgcctccgtgggcacaaaggcagcgct 181 gctggggaactcgggggaacgcgcacgtgggaaccgccgcagctccacactccaggtact 241 tcttccaaggacctaggtctctcgcccatcggaaagaaaataattctttcaagaagatca 301 gggacaactgatttgaagtctactctgtgcttctaaatccccaattctgctgaaagtgag 361 ataccctagagccctagagccccagcagcacccagccaaacccacctccaccatgggggc 421 catgactcagctgttggcaggtgtctttcttgctttccttgccctcgctaccgaaggtgg 481 ggtcctcaagaaagtcatccggcacaagcgacagagtggggtgaacgccaccctgccaga 541 agagaaccagccagtggtgtttaaccacgtttacaacatcaagctgccagtgggatccca 601 gtgttcggtggatctggagtcagccagtggggagaaagacctggcaccgccttcagagcc 661 cagcgaaagctttcaggagcacacagtggatggggaaaaccagattgtcttcacacatcg 721 catcaacatcccccgccgggcctgtggctgtgccgcagcccctgatgttaaggagctgct 781 gagcagactggaggagctggagaacctggtgtcttccctgagggagcaatgtactgcagg 841 agcaggctgctgtctccagcctgccacaggccgcttggacaccaggcccttctgtagcgg 901 tcggggcaacttcagcactgaaggatgtggctgtgtctgcgaacctggctggaaaggccc 961 caactgctctgagcccgaatgtccaggcaactgtcaccttcgaggccggtgcattgatgg 1021 gcagtgcatctgtgacgacggcttcacgggcgaggactgcagccagctggcttgccccag 1081 cgactgcaatgaccagggcaagtgcgtaaatggagtctgcatctgtttcgaaggctacgc 1141 cggggctgactgcagccgtgaaatctgcccagtgccctgcagtgaggagcacggcacatg 1201 tgtagatggcttgtgtgtgtgccacgatggctttgcaggcgatgactgcaacaagcctct 1261 gtgtctcaacaattgctacaaccgtggacgatgcgtggagaatgagtgcgtgtgtgatga 1321 gggtttcacgggcgaagactgcagtgagctcatctgccccaatgactgcttcgaccgggg 1381 ccgctgcatcaatggcacctgctactgcgaagaaggcttcacaggtgaagactgcgggaa 1441 acccacctgcccacatgcctgccacacccagggccggtgtgaggaggggcagtgtgtatg 1501 tgatgagggctttgccggtgtggactgcagcgagaagaggtgtcctgctgactgtcacaa 1561 tcgtggccgctgtgtagacgggcggtgtgagtgtgatgatggtttcactggagctgactg 1621 tggggagctcaagtgtcccaatggctgcagtggccatggccgctgtgtcaatgggcagtg 1681 tgtgtgtgatgagggctatactggggaggactgcagccagctacggtgccccaatgactg 1741 tcacagtcggggccgctgtgtcgagggcaaatgtgtatgtgagcaaggcttcaagggcta 1801 tgactgcagtgacatgagctgccctaatgactgtcaccagcacggccgctgtgtgaatgg 4441 cacagaggatctcccacagctgggagatttagccgtgtctgaggttggctgggatggcct 4501 cagactcaactggaccgcagctgacaatgcctatgagcactttgtcattcaggtgcagga 4561 ggtcaacaaagtggaggcagcccagaacctcacgttgcctggcagcctcagggctgtgga 4621 catcccgggcctcgaggctgccacgccttatagagtctccatctatggggtgatccgggg 4681 ctatagaacaccagtactctctgctgaggcctccacagccaaagaacctgaaattggaaa 4741 cttaaatgtttctgacataactcccgagagcttcaatctctcctggatggctaccgatgg 4801 gatcttcgagacctttaccattgaaattattgattccaataggttgctggagactgtgga 4861 atataatatctctggtgctgaacgaactgcccatatctcagggctaccccctagtactga 4921 ttttattgtctacctctctggacttgctcccagcatccggaccaaaaccatcagtgccac 4981 agccacgacagaggccctgccccttctggaaaacctaaccatttccgacattaatcccta 5041 cgggttcacagtttcctggatggcatcggagaatgcctttgacagctttctagtaacggt 5101 ggtggattctgggaagctgctggacccccaggaattcacactttcaggaacccagaggaa 5161 gctggagcttagaggcctcataactggcattggctatgaggttatggtctctggcttcac 5221 ccaagggcatcaaaccaagcccttgagggctgagattgttacagaagccgaaccggaagt 5281 tgacaaccttctggtttcagatgccaccccagacggtttccgtctgtcctggacagctga 5341 tgaaggggtcttcgacaattttgttctcaaaatcagagataccaaaaagcagtctgagcc 5401 actggaaataaccctacttgcccccgaacgtaccagggacataacaggtctcagagaggc 5461 tactgaatacgaaattgaactctatggaataagcaaaggaaggcgatcccagacagtcag 5521 tgctatagcaacaacagccatgggctccccaaaggaagtcattttctcagacatcactga 5581 aaattcggctactgtcagctggagggcacccacagcccaagtggagagcttccggattac 5641 ctatgtgcccattacaggaggtacaccctccatggtaactgtggacggaaccaagactca 5701 gaccaggctggtgaaactcatacctggcgtggagtaccttgtcagcatcatcgccatgaa 5761 gggctttgaggaaagtgaacctgtctcagggtcattcaccacagctctggatggcccatc 5821 tggcctggtgacagccaacatcactgactcagaagccttggccaggtggcagccagccat 5881 tgccactgtggacagttatgtcatctcctacacaggcgagaaagtgccagaaattacacg 5941 cacggtgtccgggaacacagtggagtatgctctgaccgacctcgagcctgccacggaata 6001 cacactgagaatctttgcagagaaagggccccagaagagctcaaccatcactgccaagtt 6061 cacaacagacctcgattctccaagagacttgactgctactgaggttcagtcggaaactgc 6121 cctccttacctggcgacccccccgggcatcagtcaccggttacctgctggtctatgaatc 6181 agtggatggcacagtcaaggaagtcattgtgggtccagataccacctcctacagcctggc 6241 agacctgagcccatccacccactacacagccaagatccaggcactcaatgggcccctgag 6301 gagcaatatgatccagaccatcttcaccacaattggactcctgtaccccttccccaagga 6361 ctgctcccaagcaatgctgaatggagacacgacctctggcctctacaccatttatctgaa 6421 tggtgataaggctgaggcgctggaagtcttctgtgacatgacctctgatgggggtggatg 6481 gattgtgttcctgagacgcaaaaacggacgcgagaacttctaccaaaactggaaggcata 6541 tgctgctggatttggggaccgcagagaagaattctggcttgggctggacaacctgaacaa 6601 aatcacagcccaggggcagtacgagctccgggtggacctgcgggaccatggggagacagc 6661 ctttgctgtctatgacaagttcagcgtgggagatgccaagactcgctacaagctgaaggt 6721 ggaggggtacagtgggacagcaggtgactccatggcctaccacaatggcagatccttctc 6781 cacctttgacaaggacacagattcagccatcaccaactgtgctctgtcctacaaaggggc 6841 tttctggtacaggaactgtcaccgtgtcaacctgatggggagatatggggacaataacca 6901 cagtcagggcgttaactggttccactggaagggccacgaacactcaatccagtttgctga 6961 gatgaagctgagaccaagcaacttcagaaatcttgaaggcaggcgcaaacgggcataaat 7021 tccagggaccactgggtgagagaggaataaggcccagagcgaggaaaggattttaccaaa 7081 gcatcaatacaaccagcccaaccatcggtccacacctgggcatttggtgagagtcaaagc 7141 tgaccatggatccctggggccaacggcaacagcatgggcctcacctcctctgtgatttct 7201 ttctttgcaccaaagacatcagtctccaacatgtttctgttttgttgtttgattcagcaa 7261 aaatctcccagtgacaacatcgcaatagttttttacttctcttaggtggctctgggaatg 7321 ggagaggggtaggatgtacaggggtagtttgttttagaaccagccgtattttacatgaag 7381 ctgtataattaattgtcattatttttgttagcaaagattaaatgtgtcattggaagccat 7441 ccctttttttacatttcatacaacagaaaccagaaaagcaatactgtttccattttaagg 7501 atatgattaatattattaatataataatgatgatgatgatgatgaaaactaaggattttt 7561 caagagatctttctttccaaaacatttctggacagtacctgattgtattttttttttaaa 7621 taaaagcacaagtacttttgagtttgttattttgctttgaattgttgagtctgaatttca 7681 ccaaagccaatcatttgaacaaagcggggaatgttgggataggaaaggtaagtagggata 7741 gtggtcaagtgggaggggtggaaaggagactaaagactgggagagagggaagcacttttt 7801 ttaaataaagttgaacacacttgggaaaagcttacaggccaggcctgtaatcccaacact 7861 ttgggaggccaaggtgggaggatagcttaaccccaggagtttgagaccagcctgagcaac 7921 atagtgagaacttgtctctacagaaaaaaaaaaaaaaaaaaatttaattaggcaagcgtg 7981 gtagtgcgcacctgtcgtcccagctactcaggaggctgaggtaggaaaatcactggagcc 8041 caggagttagaggttacagtgagctatgatcacactactgcactccagcctgggcaacag 8101 agggagaccctgtctctaaataaaaaaagaaaagaaaaaaaaagcttacaacttgagatt 8161 cagcatcttgctcagtatttccaagactaatagattatggtttaaaagatgcttttatac 8221 tcattttctaatgcaactcctagaaactctatgatatagttgaggtaagtattgttacca 8281 cacatgggctaagatccccagaggcagactgcctgagttcaattcttggctccaccattc 8341 ccaagttccctaacctctctatgcctcagtttcctcttctgtaaagtagggacactcata 8401 cttctcatttcagaacatttttgtgaagaataaattatgttatccatttgaggcccttag 8461 aatggtacccggtgtatattaagtgctagtacatgttagctatcatcattatcactttat 8521 atgagatggactggggttcatagaaacccaatgacttgattgtggctactactcaataaa 8581 taatagaatttggatttaaaaaaaa Osteopontin(NCBIRef.:NM_000582.2;SEQIDNO:171) 1 ctccctgtgttggtggaggatgtctgcagcagcatttaaattctgggagggcttggttgt 61 cagcagcagcaggaggaggcagagcacagcatcgtcgggaccagactcgtctcaggccag 121 ttgcagccttctcagccaaacgccgaccaaggaaaactcactaccatgagaattgcagtg 181 atttgcttttgcctcctaggcatcacctgtgccataccagttaaacaggctgattctgga 241 agttctgaggaaaagcagctttacaacaaatacccagatgctgtggccacatggctaaac 301 cctgacccatctcagaagcagaatctcctagccccacagacccttccaagtaagtccaac 361 gaaagccatgaccacatggatgatatggatgatgaagatgatgatgaccatgtggacagc 421 caggactccattgactcgaacgactctgatgatgtagatgacactgatgattctcaccag 481 tctgatgagtctcaccattctgatgaatctgatgaactggtcactgattttcccacggac 541 ctgccagcaaccgaagttttcactccagttgtccccacagtagacacatatgatggccga 601 ggtgatagtgtggtttatggactgaggtcaaaatctaagaagtttcgcagacctgacatc 661 cagtaccctgatgctacagacgaggacatcacctcacacatggaaagcgaggagttgaat 721 ggtgcatacaaggccatccccgttgcccaggacctgaacgcgccttctgattgggacagc 781 cgtgggaaggacagttatgaaacgagtcagctggatgaccagagtgctgaaacccacagc 841 cacaagcagtccagattatataagcggaaagccaatgatgagagcaatgagcattccgat 901 gtgattgatagtcaggaactttccaaagtcagccgtgaattccacagccatgaatttcac 961 agccatgaagatatgctggttgtagaccccaaaagtaaggaagaagataaacacctgaaa 1021 tttcgtatttctcatgaattagatagtgcatcttctgaggtcaattaaaaggagaaaaaa 1081 tacaatttctcactttgcatttagtcaaaagaaaaaatgctttatagcaaaatgaaagag 1141 aacatgaaatgcttctttctcagtttattggttgaatgtgtatctatttgagtctggaaa 1201 taactaatgtgtttgataattagtttagtttgtggcttcatggaaactccctgtaaacta 1261 aaagcttcagggttatgtctatgttcattctatagaagaaatgcaaactatcactgtatt 1321 ttaatatttgttattctctcatgaatagaaatttatgtagaagcaaacaaaatactttta 1381 cccacttaaaaagagaatataacattttatgtcactataatcttttgttttttaagttag 1441 tgtatattttgttgtgattatctttttgtggtgtgaataaatcttttatcttgaatgtaa 1501 taagaatttggtggtgtcaattgcttatttgttttcccacggttgtccagcaattaataa 1561 aacataaccttttttactgcctaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa Nephronectin(NCBIRef.:NM_001033047.2;SEQIDNO:172) 1 tagaagggagcgggagggggctccgggcgccgcgcagcagacctgctccggccgcgcgcc 61 tcgccgctgtcctccgggagcggcagcagtagcccgggcggcgagggctgggggttcctc 121 gagactctcagaggggcgcctcccatcggcgcccaccaccccaacctgttcctcgcgcgc 181 cactgcgctgcgccccaggacccgctgcccaacatggattttctcctggcgctggtgctg 241 gtatcctcgctctacctgcaggcggccgccgagttcgacgggaggtggcccaggcaaata 301 gtgtcatcgattggcctatgtcgttatggtgggaggattgactgctgctggggctgggct 361 cgccagtcttggggacagtgtcagcctgtgtgccaaccacgatgcaaacatggtgaatgt 421 atcgggccaaacaagtgcaagtgtcatcctggttatgctggaaaaacctgtaatcaagat 481 ctaaatgagtgtggcctgaagccccggccctgtaagcacaggtgcatgaacacttacggc 541 agctacaagtgctactgtctcaacggatatatgctcatgccggatggttcctgctcaagt 601 gccctgacctgctccatggcaaactgtcagtatggctgtgatgttgttaaaggacaaata 661 cggtgccagtgcccatcccctggcctgcagctggctcctgatgggaggacctgtgtagat 721 gttgatgaatgtgctacaggaagagcctcctgccctagatttaggcaatgtgtcaacact 781 tttgggagctacatctgcaagtgtcataaaggcttcgatctcatgtatattggaggcaaa 841 tatcaatgtcatgacatagacgaatgctcacttggtcagtatcagtgcagcagctttgct 901 cgatgttataacatacgtgggtcctacaagtgcaaatgtaaagaaggataccagggtgat 961 ggactgacttgtgtgtatatcccaaaagttatgattgaaccttcaggtccaattcatgta 1021 ccaaagggaaatggtaccattttaaagggtgacacaggaaataataattggattcctgat 1081 gttggaagtacttggtggcctccgaagacaccatatattcctcctatcattaccaacagg 1141 cctacttctaagccaacaacaagacctacaccaaagccaacaccaattcctactccacca 1201 ccaccaccacccctgccaacagagctcagaacacctctaccacctacaaccccagaaagg 1261 ccaaccaccggactgacaactatagcaccagctgccagtacacctccaggagggattaca 1321 gttgacaacagggtacagacagaccctcagaaacccagaggagatgtgttcattccacgg 1381 caaccttcaaatgacttgtttgaaatatttgaaatagaaagaggagtcagtgcagacgat 1441 gaagcaaaggatgatccaggtgttctggtacacagttgtaattttgaccatggactttgt 1501 ggatggatcagggagaaagacaatgacttgcactgggaaccaatcagggacccagcaggt 1561 ggacaatatctgacagtgtcggcagccaaagccccagggggaaaagctgcacgcttggtg 1621 ctacctctcggccgcctcatgcattcaggggacctgtgcctgtcattcaggcacaaggtg 1681 acggggctgcactctggcacactccaggtgtttgtgagaaaacacggtgcccacggagca 1741 gccctgtggggaagaaatggtggccatggctggaggcaaacacagatcaccttgcgaggg 1801 gctgacatcaagagcgtcgtcttcaaaggtgaaaaaaggcgtggtcacactggggagatt 1861 ggattagatgatgtgagcttgaaaaaaggccactgctctgaagaacgctaacaactccag 1921 aactaacaatgaactcctatgttgctctatcctctttttccaattctcatcttctctcct 1981 cttctcccttttatcaggcctaggagaagagtgggtcagtgggtcagaaggaagtctatt 2041 tggtgacccaggtttttctggcctgcttttgtgcaatcccaatgaacagtgataccctcc 2101 ttgaaatacaggggcatcgcagacacatcaaagccatctgtgggtgttgccttccatcct 2161 gtgtctctttcaggaaggcattcagcatgcgtgagccataccatcctccatcctgattac 2221 aaggtgctccttgtagcaaattatgagagtgagttacgggagcagtttttaaaagaaatc 2281 tttgcagatggctatgatgttatgtgttcggtgttgtaccatgagtagtattgacttccc 2341 ttgagatatgatgtacaatgtgcttgtgaaattgacttaccctcttcacttaagttagtt 2401 ctggcctgacctgaactctgacttttactgccattcactttataaaataagggtgtgtaa 2461 catatcaagatacatttatttttatctgttttttttttcctgttaaagacaattatgtag 2521 agtgggcacgtaatccctccttagtagtattgtgttttgtgtaaatgtgctattgatatt 2581 aagtatttacatgttccaaatatttacagactctagttgcaaggtaaagggcagcttgtg 2641 atctcaaaaaaatacatggtgaaatgtcatccagttccatgaccttatattggcagcagt 2701 aggaaattggcagaagtgttgggttgtggtaacggagtgatgaatttttttttaatggcc 2761 ttgagtttgatctctgcaaaggataggaaacctttaggaagacaagaaactgcagttaat 2821 ttagaactgtcactgtttcaagttacactttaaaaccacagcttttaccatcataacatg 2881 gctctggtaatatgtaggaagctttataaaagttttggttgattcagaaaaaggatcctg 2941 ttgcagagtgagaggaagcatagggggaaactccattggaacagattttcacacaacgtt 3001 ttaaattgatataagtttaggcagttgtagttcataacttatgttgctcatgttgtgctg 3061 tgtcaggatgggataggaagcaagtcccatgcttagaggcatgggatgtgttggaacggg 3121 atttacacacactggaggagcagggcaagttggaattctaagatccatgaacccccaact 3181 gtatttcctccctgcatattttaccaatatattaaaaaacaatgtaacttttaaaaggca 3241 tcattcctgaggtttgtcttaatttctgattaagtaatcagaatattttctgctattttt 3301 gccaggaatcacaaagatgattaaagggttggaaaaaaagatctatgatggaaaattaaa 3361 ggaactgggattattgagcctggagaagagaagactgaggggcaaaccattgatggtttt 3421 caagtatatgaagggttggcacagagagggtggcgaccagctgttctccatatgcactaa 3481 gaatagaacaagaggaaactggcttagactagagtataagggagcatttcttggcagggg 3541 ccattgttagaatacttcataaaaaaagaagtgtgaaaatctcagtatctctctctcttt 3601 ctaaaaaattagataaaaatttgtctatttaagatggttaaagatgttcttacccaagga 3661 aaagtaacaaattatagaatttcccaaaagatgttttgatcctactagtagtatgcagtg 3721 aaaatctttagaactaaataatttggacaaggcttaatttaggcatttccctcttgacct 3781 cctaatggagagggattgaaaggggaagagcccaccaaatgctgagctcactgaaatatc 3841 tctcccttatggcaatcctagcagtattaaagaaaaaaggaaactatttattccaaatga 3901 gagtatgatggacagatattttagtatctcagtaatgtcctagtgtggcggtggttttca 3961 atgtttcttcatgttaaaggtataagcctttcatttgttcaatggatgatgtttcagatt 4021 tttttttttttaagagatccttcaaggaacacagttcagagagattttcatcgggtgcat 4081 tctctctgcttcgtgtgtgacaagttatcttggctgctgagaaagagtgccctgccccac 4141 accggcagacctttccttcacctcatcagtatgattcagtttctcttatcaattggactc 4201 tcccaggttccacagaacagtaatattttttgaacaataggtacaatagaaggtcttctg 4261 tcatttaacctggtaaaggcagggctggagggggaaaataaatcattaagcctttgagta 4321 acggcagaatatatggctgtagatccatttttaatggttcatttcctttatggtcatata 4381 actgcacagctgaagatgaaaggggaaaataaatgaaaattttacttttcgatgccaatg 4441 atacattgcactaaactgatggaagaagttatccaaagtactgtataacatcttgtttat 4501 tatttaatgttttctaaaataaaaaatgttagtggttttccaaatggcctaataaaaaca 4561 attatttgtaaataaaaacactgttagtaata Angiostatin(PLG)(NCBIRef.:NM_000301.3;SEQIDNO:173) 1 gaatcattaacttaatttgactatctggtttgtggatgcgtttactctcatgtaagtcaa 61 caacatcctgggattgggacccactttctgggcactgctggccagtcccaaaatggaaca 121 taaggaagtggttcttctacttcttttatttctgaaatcaggtcaaggagagcctctgga 181 tgactatgtgaatacccagggggcttcactgttcagtgtcactaagaagcagctgggagc 241 aggaagtatagaagaatgtgcagcaaaatgtgaggaggacgaagaattcacctgcagggc 301 attccaatatcacagtaaagagcaacaatgtgtgataatggctgaaaacaggaagtcctc 361 cataatcattaggatgagagatgtagttttatttgaaaagaaagtgtatctctcagagtg 421 caagactgggaatggaaagaactacagagggacgatgtccaaaacaaaaaatggcatcac 481 ctgtcaaaaatggagttccacttctccccacagacctagattctcacctgctacacaccc 541 ctcagagggactggaggagaactactgcaggaatccagacaacgatccgcaggggccctg 601 gtgctatactactgatccagaaaagagatatgactactgcgacattcttgagtgtgaaga 661 ggaatgtatgcattgcagtggagaaaactatgacggcaaaatttccaagaccatgtctgg 721 actggaatgccaggcctgggactctcagagcccacacgctcatggatacattccttccaa 781 atttccaaacaagaacctgaagaagaattactgtcgtaaccccgatagggagctgcggcc 841 ttggtgtttcaccaccgaccccaacaagcgctgggaactttgtgacatcccccgctgcac 901 aacacctccaccatcttctggtcccacctaccagtgtctgaagggaacaggtgaaaacta 961 tcgcgggaatgtggctgttaccgtgtccgggcacacctgtcagcactggagtgcacagac 1021 ccctcacacacataacaggacaccagaaaacttcccctgcaaaaatttggatgaaaacta 1081 ctgccgcaatcctgacggaaaaagggccccatggtgccatacaaccaacagccaagtgcg 1141 gtgggagtactgtaagataccgtcctgtgactcctccccagtatccacggaacaattggc 1201 tcccacagcaccacctgagctaacccctgtggtccaggactgctaccatggtgatggaca 1261 gagctaccgaggcacatcctccaccaccaccacaggaaagaagtgtcagtcttggtcatc 1321 tatgacaccacaccggcaccagaagaccccagaaaactacccaaatgctggcctgacaat 1381 gaactactgcaggaatccagatgccgataaaggcccctggtgttttaccacagaccccag 1441 cgtcaggtgggagtactgcaacctgaaaaaatgctcaggaacagaagcgagtgttgtagc 1501 acctccgcctgttgtcctgcttccagatgtagagactccttccgaagaagactgtatgtt 1561 tgggaatgggaaaggataccgaggcaagagggcgaccactgttactgggacgccatgcca 1621 ggactgggctgcccaggagccccatagacacagcattttcactccagagacaaatccacg 1681 ggcgggtctggaaaaaaattactgccgtaaccctgatggtgatgtaggtggtccctggtg 1741 ctacacgacaaatccaagaaaactttacgactactgtgatgtccctcagtgtgcggcccc 1801 ttcatttgattgtgggaagcctcaagtggagccgaagaaatgtcctggaagggttgtagg 1861 ggggtgtgtggcccacccacattcctggccctggcaagtcagtcttagaacaaggtttgg 1921 aatgcacttctgtggaggcaccttgatatccccagagtgggtgttgactgctgcccactg 1981 cttggagaagtccccaaggccttcatcctacaaggtcatcctgggtgcacaccaagaagt 2041 gaatctcgaaccgcatgttcaggaaatagaagtgtctaggctgttcttggagcccacacg 2101 aaaagatattgccttgctaaagctaagcagtcctgccgtcatcactgacaaagtaatccc 2161 agcttgtctgccatccccaaattatgtggtcgctgaccggaccgaatgtttcatcactgg 2221 ctggggagaaacccaaggtacttttggagctggccttctcaaggaagcccagctccctgt 2281 gattgagaataaagtgtgcaatcgctatgagtttctgaatggaagagtccaatccaccga 2341 actctgtgctgggcatttggccggaggcactgacagttgccagggtgacagtggaggtcc 2401 tctggtttgcttcgagaaggacaaatacattttacaaggagtcacttcttggggtcttgg 2461 ctgtgcacgccccaataagcctggtgtctatgttcgtgtttcaaggtttgttacttggat 2521 tgagggagtgatgagaaataattaattggacgggagacagagtgacgcactgactcacct 2581 agaggctggaacgtgggtagggatttagcatgctggaaataactggcagtaatcaaacga 2641 agacactgtccccagctaccagctacgccaaacctcggcattttttgtgttattttctga 2701 ctgctggattctgtagtaaggtgacatagctatgacatttgttaaaaataaactctgtac 2761 ttaactttgatttgagtaaattttggttttggtcttcaacattttcatgctctttgttca 2821 ccccaccaatttttaaatgggcagatggggggatttagctgcttttgataaggaacagct 2881 gcacaaaggactgagcaggctgcaaggtcacagaggggagagccaagaagttgtccacgc 2941 atttacctcatcagctaacgagggcttgacatgcatttttactgtctttattcctgacac 3001 tgagatgaatgttttcaaagctgcaacatgtatggggagtcatgcaaaccgattctgtta 3061 ttgggaatgaaatctgtcaccgactgcttgacttgagcccaggggacacggagcagagag 3121 ctgtatatgatggagtgaaccggtccatggatgtgtaacacaagaccaactgagagtctg 3181 aatgttattctggggcacacgtgagtctaggattggtgccaagagcatgtaaatgaacaa 3241 caagcaaatattgaaggtggaccacttatttcccattgctaattgcctgcccggttttga 3301 aacagtctgcagtacacacggtcacaggagaatgacctgtgggagagatacatgtttaga 3361 aggaagagaaaggacaaaggcacacgttttaccatttaaaatattgttaccaaacaaaaa 3421 tatccattcaaaatacaatttaacaatgcaacagtcatcttacagcagagaaatgcagag 3481 aaaagcaaaactgcaagtgactgtgaataaagggtgaatgtagtctcaaatcctcaaa TissuetransglutaminasefactorXIII(F13A1)(NCBIRef.:NM_000129.3; SEQIDNO:174) 1 atttaagagccaactgtcttgtctttcccgagtccgtttgaggaagtccccgaggcgcac 61 agagcaagcccacgcgagggcacctctggaggggagcgcctgcaggaccttgtaaagtca 121 aaaatgtcagaaacttccaggaccgcctttggaggcagaagagcagttccacccaataac 181 tctaatgcagcggaagatgacctgcccacagtggagcttcagggcgtggtgccccggggc 241 gtcaacctgcaagagtttcttaatgtcacgagcgttcacctgttcaaggagagatgggac 301 actaacaaggtggaccaccacactgacaagtatgaaaacaacaagctgattgtccgcaga 361 gggcagtctttctatgtgcagattgacttcagtcgtccatatgaccccagaagggatctc 421 ttcagggtggaatacgtcattggtcgctacccacaggagaacaagggaacctacatccca 481 gtgcctatagtctcagagttacaaagtggaaagtggggggccaagattgtcatgagagag 541 gacaggtctgtgcggctgtccatccagtcttcccccaaatgtattgtggggaaattccgc 601 atgtatgttgctgtctggactccctatggcgtacttcgaaccagtcgaaacccagaaaca 661 gacacgtacattctcttcaatccttggtgtgaagatgatgctgtgtatctggacaatgag 721 aaagaaagagaagagtatgtcctgaatgacatcggggtaattttttatggagaggtcaat 781 gacatcaagaccagaagctggagctatggtcagtttgaagatggcatcctggacacttgc 841 ctgtatgtgatggacagagcacaaatggacctctctggaagagggaatcccatcaaagtc 901 agccgtgtggggtctgcaatggtgaatgccaaagatgacgaaggtgtcctcgttggatcc 961 tgggacaatatctatgcctatggcgtccccccatcggcctggactggaagcgttgacatt 1021 ctattggaataccggagctctgagaatccagtccggtatggccaatgctgggtttttgct 1081 ggtgtctttaacacatttttacgatgccttggaataccagcaagaattgttaccaattat 1141 ttctctgcccatgataatgatgccaatttgcaaatggacatcttcctggaagaagatggg 1201 aacgtgaattccaaactcaccaaggattcagtgtggaactaccactgctggaatgaagca 1261 tggatgacaaggcctgaccttcctgttggatttggaggctggcaagctgtggacagcacc 1321 ccccaggaaaatagcgatggcatgtatcggtgtggccccgcctcggttcaagccatcaag 1381 cacggccatgtctgcttccaatttgatgcaccttttgtttttgcagaggtcaacagcgac 1441 ctcatttacattacagctaagaaagatggcactcatgtggtggaaaatgtggatgccacc 1501 cacattgggaaattaattgtgaccaaacaaattggaggagatggcatgatggatattact 1561 gatacttacaaattccaagaaggtcaagaagaagagagattggccctagaaactgccctg 1621 atgtacggagctaaaaagcccctcaacacagaaggtgtcatgaaatcaaggtccaacgtt 1681 gacatggactttgaagtggaaaatgctgtgctgggaaaagacttcaagctctccatcacc 1741 ttccggaacaacagccacaaccgttacaccatcacagcttatctctcagccaacatcacc 1801 ttctacaccggggtcccgaaggcagaattcaagaaggagacgttcgacgtgacgctggag 1861 cccttgtccttcaagaaagaggcggtgctgatccaagccggcgagtacatgggtcagctg 1921 ctggaacaagcgtccctgcacttctttgtcacagctcgcatcaatgagaccagggatgtt 1981 ctggccaagcaaaagtccaccgtgctaaccatccctgagatcatcatcaaggtccgtggc 2041 actcaggtagttggttctgacatgactgtgacagttgagtttaccaatcctttaaaagaa 2101 accctgcgaaatgtctgggtacacctggatggtcctggagtaacaagaccaatgaagaag 2161 atgttccgtgaaatccggcccaactccaccgtgcagtgggaagaagtgtgccggccctgg 2221 gtctctgggcatcggaagctgatagccagcatgagcagtgactccctgagacatgtgtat 2281 ggcgagctggacgtgcagattcaaagacgaccttccatgtgaatgcacaggaagctgaga 2341 tgaaccctggcatttggcctcttgtagtcttggctaaggaaattctaacgcaaaaatagc 2401 tcttgctttgacttaggtgtgaagacccagacaggactgcagagggctccagagtggaga 2461 tcccacatatttcaaaaacatgcttttccaaacccaggctattcggcaaggaagttagtt 2521 tttaatctctccaccttccaaagagtgctaagcattagctttaattaagctctcatagct 2581 cataagagtaacagtcatcatttatcatcacaaatggctacatctccaaatatcagtggg 2641 ctctcttaccagggagatttgctcaatacctggcctcatttaaaacaagacttcagattc 2701 cccactcagccttttgggaataatagcacatgatttgggctctagaattccagtcccctt 2761 tctcggggtcaggttctaccctccatgtgagaatatttttcccaggactagagcacaaca 2821 taatttttatttttggcaaagccagaaaaagatctttcattttgcacctgcagccaagca 2881 aatgcctgccaaattttagatttaccttgttagaagaggtggccccatattaacaaattg 2941 catttgtgggaaacttaaccacctacaaggagataagaaagcaggtgcaacactcaagtc 3001 tattgaataatgtagttttgtgatgcattttatagaatgtgtcacactgtggcctgatca 3061 gcaggagccaatatcccttactttaaccctttctgggatgcaatactaggaagtaaagtg 3121 aagaatttatctctttagttagtgattatatttcacccatctctcaggaatcatctcctt 3181 tgcagaatgatgcaggttcaggtcccctttcagagatataataagcccaacaagttgaag 3241 aagctggcggatctagtgaccagatatatagaaggactgcagccactgattctctcttgt 3301 ccttcacatcacccatgttgagacctcagcttggcactcaggtgctgaagggtaatatgg 3361 actcagccttgcaaatagccagtgctagttctgacccaaccacagaggatgctgacatca 3421 tttgtattatgttccaaggctactacagagaaggctgcctgctatgtatttgcaaggctg 3481 atttatggtcagaatttccctctgatatgtctagggtgtgatttaggtcagtagactgtg 3541 attcttagcaaaaaatgaacagtgataagtatactgggggcaaaatcagaatggaatgct 3601 ctggtctatataaccacatttctaagcctttgagactgttcctgagccttcagcactaac 3661 ctatgagggtgagctggtcccctctatatatacatcatacttaactttactaagtaatct 3721 cacagcatttgccaagtctcccaatatccaattttaaaatgaaatgcattttgctagaca 3781 gttaaactggcttaacttagtatattattattaattacaatgtaatagaagcttaaaata 3841 aagttaaactgattatatttgca VonWillebrandFactor(NCBIRef.:NM_000552.4;SEQIDNO:175) 1 gtggcagctcacagctattgtggtgggaaagggagggtggttggtggatgtcacagcttg 61 ggctttatctcccccagcagtggggactccacagcccctgggctacataacagcaagaca 121 gtccggagctgtagcagacctgattgagcctttgcagcagctgagagcatggcctagggt 181 gggcggcaccattgtccagcagctgagtttcccagggaccttggagatagccgcagccct 241 catttgcaggggaagatgattcctgccagatttgccggggtgctgcttgctctggccctc 301 attttgccagggaccctttgtgcagaaggaactcgcggcaggtcatccacggcccgatgc 361 agccttttcggaagtgacttcgtcaacacctttgatgggagcatgtacagctttgcggga 421 tactgcagttacctcctggcagggggctgccagaaacgctccttctcgattattggggac 481 ttccagaatggcaagagagtgagcctctccgtgtatcttggggaattttttgacatccat 541 ttgtttgtcaatggtaccgtgacacagggggaccaaagagtctccatgccctatgcctcc 601 aaagggctgtatctagaaactgaggctgggtactacaagctgtccggtgaggcctatggc 661 tttgtggccaggatcgatggcagcggcaactttcaagtcctgctgtcagacagatacttc 721 aacaagacctgcgggctgtgtggcaactttaacatctttgctgaagatgactttatgacc 781 caagaagggaccttgacctcggacccttatgactttgccaactcatgggctctgagcagt 841 ggagaacagtggtgtgaacgggcatctcctcccagcagctcatgcaacatctcctctggg 901 gaaatgcagaagggcctgtgggagcagtgccagcttctgaagagcacctcggtgtttgcc 961 cgctgccaccctctggtggaccccgagccttttgtggccctgtgtgagaagactttgtgt 1021 gagtgtgctggggggctggagtgcgcctgccctgccctcctggagtacgcccggacctgt 1081 gcccaggagggaatggtgctgtacggctggaccgaccacagcgcgtgcagcccagtgtgc 1141 cctgctggtatggagtataggcagtgtgtgtccccttgcgccaggacctgccagagcctg 1201 cacatcaatgaaatgtgtcaggagcgatgcgtggatggctgcagctgccctgagggacag 1261 ctcctggatgaaggcctctgcgtggagagcaccgagtgtccctgcgtgcattccggaaag 1321 cgctaccctcccggcacctccctctctcgagactgcaacacctgcatttgccgaaacagc 1381 cagtggatctgcagcaatgaagaatgtccaggggagtgccttgtcacaggtcaatcacac 1441 ttcaagagctttgacaacagatacttcaccttcagtgggatctgccagtacctgctggcc 1501 cgggattgccaggaccactccttctccattgtcattgagactgtccagtgtgctgatgac 1561 cgcgacgctgtgtgcacccgctccgtcaccgtccggctgcctggcctgcacaacagcctt 1621 gtgaaactgaagcatggggcaggagttgccatggatggccaggacgtccagctccccctc 1681 ctgaaaggtgacctccgcatccagcatacagtgacggcctccgtgcgcctcagctacggg 1741 gaggacctgcagatggactgggatggccgcgggaggctgctggtgaagctgtcccccgtc 1801 tatgccgggaagacctgcggcctgtgtgggaattacaatggcaaccagggcgacgacttc 1861 cttaccccctctgggctggcggagccccgggtggaggacttcgggaacgcctggaagctg 1921 cacggggactgccaggacctgcagaagcagcacagcgatccctgcgccctcaacccgcgc 1981 atgaccaggttctccgaggaggcgtgcgcggtcctgacgtcccccacattcgaggcctgc 2041 catcgtgccgtcagcccgctgccctacctgcggaactgccgctacgacgtgtgctcctgc 2101 tcggacggccgcgagtgcctgtgcggcgccctggccagctatgccgcggcctgcgcgggg 2161 agaggcgtgcgcgtcgcgtggcgcgagccaggccgctgtgagctgaactgcccgaaaggc 2221 caggtgtacctgcagtgcgggaccccctgcaacctgacctgccgctctctctcttacccg 2281 gatgaggaatgcaatgaggcctgcctggagggctgcttctgccccccagggctctacatg 2341 gatgagaggggggactgcgtgcccaaggcccagtgcccctgttactatgacggtgagatc 2401 ttccagccagaagacatcttctcagaccatcacaccatgtgctactgtgaggatggcttc 2461 atgcactgtaccatgagtggagtccccggaagcttgctgcctgacgctgtcctcagcagt 2521 cccctgtctcatcgcagcaaaaggagcctatcctgtcggccccccatggtcaagctggtg 2581 tgtcccgctgacaacctgcgggctgaagggctcgagtgtaccaaaacgtgccagaactat 2641 gacctggagtgcatgagcatgggctgtgtctctggctgcctctgccccccgggcatggtc 2701 cggcatgagaacagatgtgtggccctggaaaggtgtccctgcttccatcagggcaaggag 2761 tatgcccctggagaaacagtgaagattggctgcaacacttgtgtctgtcgggaccggaag 2821 tggaactgcacagaccatgtgtgtgatgccacgtgctccacgatcggcatggcccactac 2881 ctcaccttcgacgggctcaaatacctgttccccggggagtgccagtacgttctggtgcag 2941 gattactgcggcagtaaccctgggacctttcggatcctagtggggaataagggatgcagc 3001 cacccctcagtgaaatgcaagaaacgggtcaccatcctggtggagggaggagagattgag 3061 ctgtttgacggggaggtgaatgtgaagaggcccatgaaggatgagactcactttgaggtg 3121 gtggagtctggccggtacatcattctgctgctgggcaaagccctctccgtggtctgggac 3181 cgccacctgagcatctccgtggtcctgaagcagacataccaggagaaagtgtgtggcctg 3241 tgtgggaattttgatggcatccagaacaatgacctcaccagcagcaacctccaagtggag 3301 gaagaccctgtggactttgggaactcctggaaagtgagctcgcagtgtgctgacaccaga 3361 aaagtgcctctggactcatcccctgccacctgccataacaacatcatgaagcagacgatg 3421 gtggattcctcctgtagaatccttaccagtgacgtcttccaggactgcaacaagctggtg 3481 gaccccgagccatatctggatgtctgcatttacgacacctgctcctgtgagtccattggg 3541 gactgcgcctgcttctgcgacaccattgctgcctatgcccacgtgtgtgcccagcatggc 3601 aaggtggtgacctggaggacggccacattgtgcccccagagctgcgaggagaggaatctc 3661 cgggagaacgggtatgagtgtgagtggcgctataacagctgtgcacctgcctgtcaagtc 3721 acgtgtcagcaccctgagccactggcctgccctgtgcagtgtgtggagggctgccatgcc 3781 cactgccctccagggaaaatcctggatgagcttttgcagacctgcgttgaccctgaagac 3841 tgtccagtgtgtgaggtggctggccggcgttttgcctcaggaaagaaagtcaccttgaat 3901 cccagtgaccctgagcactgccagatttgccactgtgatgttgtcaacctcacctgtgaa 3961 gcctgccaggagccgggaggcctggtggtgcctcccacagatgccccggtgagccccacc 4021 actctgtatgtggaggacatctcggaaccgccgttgcacgatttctactgcagcaggcta 4081 ctggacctggtcttcctgctggatggctcctccaggctgtccgaggctgagtttgaagtg 4141 ctgaaggcctttgtggtggacatgatggagcggctgcgcatctcccagaagtgggtccgc 4201 gtggccgtggtggagtaccacgacggctcccacgcctacatcgggctcaaggaccggaag 4261 cgaccgtcagagctgcggcgcattgccagccaggtgaagtatgcgggcagccaggtggcc 4321 tccaccagcgaggtcttgaaatacacactgttccaaatcttcagcaagatcgaccgccct 4381 gaagcctcccgcatcaccctgctcctgatggccagccaggagccccaacggatgtcccgg 4441 aactttgtccgctacgtccagggcctgaagaagaagaaggtcattgtgatcccggtgggc 4501 attgggccccatgccaacctcaagcagatccgcctcatcgagaagcaggcccctgagaac 4561 aaggccttcgtgctgagcagtgtggatgagctggagcagcaaagggacgagatcgttagc 4621 tacctctgtgaccttgcccctgaagcccctcctcctactctgccccccgacatggcacaa 4681 gtcactgtgggcccggggctcttgggggtttcgaccctggggcccaagaggaactccatg 4741 gttctggatgtggcgttcgtcctggaaggatcggacaaaattggtgaagccgacttcaac 4801 aggagcaaggagttcatggaggaggtgattcagcggatggatgtgggccaggacagcatc 4861 cacgtcacggtgctgcagtactcctacatggtgactgtggagtaccccttcagcgaggca 4921 cagtccaaaggggacatcctgcagcgggtgcgagagatccgctaccagggcggcaacagg 4981 accaacactgggctggccctgcggtacctctctgaccacagcttcttggtcagccagggt 5041 gaccgggagcaggcgcccaacctggtctacatggtcaccggaaatcctgcctctgatgag 5101 atcaagaggctgcctggagacatccaggtggtgcccattggagtgggccctaatgccaac 5161 gtgcaggagctggagaggattggctggcccaatgcccctatcctcatccaggactttgag 5221 acgctcccccgagaggctcctgacctggtgctgcagaggtgctgctccggagaggggctg 5281 cagatccccaccctctcccctgcacctgactgcagccagcccctggacgtgatccttctc 5341 ctggatggctcctccagtttcccagcttcttattttgatgaaatgaagagtttcgccaag 5401 gctttcatttcaaaagccaatatagggcctcgtctcactcaggtgtcagtgctgcagtat 5461 ggaagcatcaccaccattgacgtgccatggaacgtggtcccggagaaagcccatttgctg 5521 agccttgtggacgtcatgcagcgggagggaggccccagccaaatcggggatgccttgggc 5581 tttgctgtgcgatacttgacttcagaaatgcatggtgccaggccgggagcctcaaaggcg 5641 gtggtcatcctggtcacggacgtctctgtggattcagtggatgcagcagctgatgccgcc 5701 aggtccaacagagtgacagtgttccctattggaattggagatcgctacgatgcagcccag 5761 ctacggatcttggcaggcccagcaggcgactccaacgtggtgaagctccagcgaatcgaa 5821 gacctccctaccatggtcaccttgggcaattccttcctccacaaactgtgctctggattt 5881 gttaggatttgcatggatgaggatgggaatgagaagaggcccggggacgtctggaccttg 5941 ccagaccagtgccacaccgtgacttgccagccagatggccagaccttgctgaagagtcat 6001 cgggtcaactgtgaccgggggctgaggccttcgtgccctaacagccagtcccctgttaaa 6061 gtggaagagacctgtggctgccgctggacctgcccctgcgtgtgcacaggcagctccact 6121 cggcacatcgtgacctttgatgggcagaatttcaagctgactggcagctgttcttatgtc 6181 ctatttcaaaacaaggagcaggacctggaggtgattctccataatggtgcctgcagccct 6241 ggagcaaggcagggctgcatgaaatccatcgaggtgaagcacagtgccctctccgtcgag 6301 ctgcacagtgacatggaggtgacggtgaatgggagactggtctctgttccttacgtgggt 6361 gggaacatggaagtcaacgtttatggtgccatcatgcatgaggtcagattcaatcacctt 6421 ggtcacatcttcacattcactccacaaaacaatgagttccaactgcagctcagccccaag 6481 acttttgcttcaaagacgtatggtctgtgtgggatctgtgatgagaacggagccaatgac 6541 ttcatgctgagggatggcacagtcaccacagactggaaaacacttgttcaggaatggact 6601 gtgcagcggccagggcagacgtgccagcccatcctggaggagcagtgtcttgtccccgac 6661 agctcccactgccaggtcctcctcttaccactgtttgctgaatgccacaaggtcctggct 6721 ccagccacattctatgccatctgccagcaggacagttgccaccaggagcaagtgtgtgag 6781 gtgatcgcctcttatgcccacctctgtcggaccaacggggtctgcgttgactggaggaca 6841 cctgatttctgtgctatgtcatgcccaccatctctggtctacaaccactgtgagcatggc 6901 tgtccccggcactgtgatggcaacgtgagctcctgtggggaccatccctccgaaggctgt 6961 ttctgccctccagataaagtcatgttggaaggcagctgtgtccctgaagaggcctgcact 7021 cagtgcattggtgaggatggagtccagcaccagttcctggaagcctgggtcccggaccac 7081 cagccctgtcagatctgcacatgcctcagcgggcggaaggtcaactgcacaacgcagccc 7141 tgccccacggccaaagctcccacgtgtggcctgtgtgaagtagcccgcctccgccagaat 7201 gcagaccagtgctgccccgagtatgagtgtgtgtgtgacccagtgagctgtgacctgccc 7261 ccagtgcctcactgtgaacgtggcctccagcccacactgaccaaccctggcgagtgcaga 7321 cccaacttcacctgcgcctgcaggaaggaggagtgcaaaagagtgtccccaccctcctgc 7381 cccccgcaccgtttgcccacccttcggaagacccagtgctgtgatgagtatgagtgtgcc 7441 tgcaactgtgtcaactccacagtgagctgtccccttgggtacttggcctcaactgccacc 7501 aatgactgtggctgtaccacaaccacctgccttcccgacaaggtgtgtgtccaccgaagc 7561 accatctaccctgtgggccagttctgggaggagggctgcgatgtgtgcacctgcaccgac 7621 atggaggatgccgtgatgggcctccgcgtggcccagtgctcccagaagccctgtgaggac 7681 agctgtcggtcgggcttcacttacgttctgcatgaaggcgagtgctgtggaaggtgcctg 7741 ccatctgcctgtgaggtggtgactggctcaccgcggggggactcccagtcttcctggaag 7801 agtgtcggctcccagtgggcctccccggagaacccctgcctcatcaatgagtgtgtccga 7861 gtgaaggaggaggtctttatacaacaaaggaacgtctcctgcccccagctggaggtccct 7921 gtctgcccctcgggctttcagctgagctgtaagacctcagcgtgctgcccaagctgtcgc 7981 tgtgagcgcatggaggcctgcatgctcaatggcactgtcattgggcccgggaagactgtg 8041 atgatcgatgtgtgcacgacctgccgctgcatggtgcaggtgggggtcatctctggattc 8101 aagctggagtgcaggaagaccacctgcaacccctgccccctgggttacaaggaagaaaat 8161 aacacaggtgaatgttgtgggagatgtttgcctacggcttgcaccattcagctaagagga 8221 ggacagatcatgacactgaagcgtgatgagacgctccaggatggctgtgatactcacttc 8281 tgcaaggtcaatgagagaggagagtacttctgggagaagagggtcacaggctgcccaccc 8341 tttgatgaacacaagtgtctggctgagggaggtaaaattatgaaaattccaggcacctgc 8401 tgtgacacatgtgaggagcctgagtgcaacgacatcactgccaggctgcagtatgtcaag 8461 gtgggaagctgtaagtctgaagtagaggtggatatccactactgccagggcaaatgtgcc 8521 agcaaagccatgtactccattgacatcaacgatgtgcaggaccagtgctcctgctgctct 8581 ccgacacggacggagcccatgcaggtggccctgcactgcaccaatggctctgttgtgtac 8641 catgaggttctcaatgccatggagtgcaaatgctcccccaggaagtgcagcaagtgaggc 8701 tgctgcagctgcatgggtgcctgctgctgcctgccttggcctgatggccaggccagagtg 8761 ctgccagtcctctgcatgttctgctcttgtgcccttctgagcccacaataaaggctgagc 8821 tcttatcttgcaaaaggc ADAM2(NCBIRef.:NM_001278113.1;SEQIDNO:176) 1 gcctacctcttccaggctgcgtggccggggcgtcatctcgcgcttccaactgccctgtaa 61 ccaccaactgccattattccggctgggacccaggacttcaagccatgtggcgcgtcttgt 121 ttctgctcagcgggctcggcgggctgcggatggacagtaattttgatagtttacctgtgc 181 aaattacagttccggagaaaatacggtcaataataaaggaaggaattgaatcgcaggcat 241 cctacaaaattgtaattgaagggaaaccatatactgtgaatttaatgcaaaaaaactttt 301 taccccataattttagagtttacagttatagtggcacaggaattatgaaaccacttgacc 361 aagattttcagaatttctgccactaccaagggtatattgaaggttatccaaaatctgtgg 421 tgatggttagcacatgtactggactcaggggcgtactacagtttgaaaatgttagttatg 481 gaatagaacccctggagtcttcagttggctttgaacatgtaatttaccaagtaaaacata 541 agaaagcagatgtttccttatataatgagaaggatattgaatcaagagatctgtccttta 601 aattacaaagcgtagagtataatcatatggggtctgatacaactgttgtcgctcaaaaag 661 ttttccagttgattggattgacgaatgctatttttgtttcatttaatattacaattattc 721 tgtcttcattggagctttggatagatgaaaataaaattgcaaccactggagaagctaatg 781 agttattacacacatttttaagatggaaaacatcttatcttgttttacgtcctcatgatg 841 tggcatttttacttgtttacagagaaaagtcaaattatgttggtgcaacctttcaaggga 901 agatgtgtgatgcaaactatgcaggaggtgttgttctgcaccccagaaccataagtctgg 961 aatcacttgcagttattttagctcaattattgagccttagtatggggatcacttatgatg 1021 acattaacaaatgccagtgctcaggagctgtctgcattatgaatccagaagcaattcatt 1081 tcagtggtgtgaagatctttagtaactgcagcttcgaagactttgcacattttatttcaa 1141 agcagaagtcccagtgtcttcacaatcagcctcgcttagatccttttttcaaacagcaag 1201 cagtgtgtggtaatgcaaagctggaagcaggagaggagtgtgactgtgggactgaacagg 1261 attgtgcccttattggagaaacatgctgtgatattgccacatgtagatttaaagccggtt 1321 caaactgtgctgaaggaccatgctgcgaaaactgtctatttatgtcaaaagaaagaatgt 1381 gtaggccttcctttgaagaatgcgacctccctgaatattgcaatggatcatctgcatcat 1441 gcccagaaaaccactatgttcagactgggcatccgtgtggactgaatcaatggatctgta 1501 tagatggagtttgtatgagtggggataaacaatgtacagacacatttggcaaagaagtag 1561 agtttggcccttcagaatgttattctcaccttaattcaaagactgatgtatctggaaact 1621 gtggtataagtgattcaggatacacacagtgtgaagctgacaatctgcagtgcggaaaat 1681 taatatgtaaatatgtaggtaaatttttattacaaattccaagagccactattatttatg 1741 ccaacataagtggacatctctgcattgctgtggaatttgccagtgatcatgcagacagcc 1801 aaaagatgtggataaaagatggaacttcttgtggttcaaataaggtttgcaggaatcaaa 1861 gatgtgtgagttcttcatacttgggttatgattgtactactgacaaatgcaatgatagag 1921 gtgtatgcaataacaaaaagcactgtcactgtagtgcttcatatttacctccagattgct 1981 cagttcaatcagatctatggcctggtgggagtattgacagtggcaattttccacctgtag 2041 ctataccagccagactccctgaaaggcgctacattgagaacatttaccattccaaaccaa 2101 tgagatggccatttttcttattcattcctttctttattattttctgtgtactgattgcta 2161 taatggtgaaagttaatttccaaaggaaaaaatggagaactgaggactattcaagcgatg 2221 agcaacctgaaagtgagagtgaacctaaagggtagtctggacaacagagatgccatgata 2281 tcacttcttctagagtaattatctgtgatggatggacacaaaaaaatggaaagaaaagaa 2341 tgtacattacctggtttcctgggattcaaacctgcatattgtgattttaatttgaccaga 2401 aaatatgatatatatgtataatttcacagataatttacttatttaaaaatgcatgataat 2461 gagttttacattacaaatttctgtttttttaaagttatcttacgctatttctgttggtta 2521 gtagacactaattctgtcagtaggggcatggtataaggaaatatcataatgtaatgaggt 2581 ggtactatgattaaaagccactgttacatttcaaaaaaaaaaaaaaa ICAM1(NCBIRef.:NM_000201.2;SEQIDNO:177) 1 caagcttagcctggccgggaaacgggaggcgtggaggccgggagcagcccccggggtcat 61 cgccctgccaccgccgcccgattgctttagcttggaaattccggagctgaagcggccagc 121 gagggaggatgaccctctcggcccgggcaccctgtcagtccggaaataactgcagcattt 181 gttccggaggggaaggcgcgaggtttccgggaaagcagcaccgccccttggcccccaggt 241 ggctagcgctataaaggatcacgcgccccagtcgacgctgagctcctctgctactcagag 301 ttgcaacctcagcctcgctatggctcccagcagcccccggcccgcgctgcccgcactcct 361 ggtcctgctcggggctctgttcccaggacctggcaatgcccagacatctgtgtccccctc 421 aaaagtcatcctgccccggggaggctccgtgctggtgacatgcagcacctcctgtgacca 481 gcccaagttgttgggcatagagaccccgttgcctaaaaaggagttgctcctgcctgggaa 541 caaccggaaggtgtatgaactgagcaatgtgcaagaagatagccaaccaatgtgctattc 601 aaactgccctgatgggcagtcaacagctaaaaccttcctcaccgtgtactggactccaga 661 acgggtggaactggcacccctcccctcttggcagccagtgggcaagaaccttaccctacg 721 ctgccaggtggagggtggggcaccccgggccaacctcaccgtggtgctgctccgtgggga 781 gaaggagctgaaacgggagccagctgtgggggagcccgctgaggtcacgaccacggtgct 841 ggtgaggagagatcaccatggagccaatttctcgtgccgcactgaactggacctgcggcc 901 ccaagggctggagctgtttgagaacacctcggccccctaccagctccagacctttgtcct 961 gccagcgactcccccacaacttgtcagcccccgggtcctagaggtggacacgcaggggac 1021 cgtggtctgttccctggacgggctgttcccagtctcggaggcccaggtccacctggcact 1081 gggggaccagaggttgaaccccacagtcacctatggcaacgactccttctcggccaaggc 1141 ctcagtcagtgtgaccgcagaggacgagggcacccagcggctgacgtgtgcagtaatact 1201 ggggaaccagagccaggagacactgcagacagtgaccatctacagctttccggcgcccaa 1261 cgtgattctgacgaagccagaggtctcagaagggaccgaggtgacagtgaagtgtgaggc 1321 ccaccctagagccaaggtgacgctgaatggggttccagcccagccactgggcccgagggc 1381 ccagctcctgctgaaggccaccccagaggacaacgggcgcagcttctcctgctctgcaac 1441 cctggaggtggccggccagcttatacacaagaaccagacccgggagcttcgtgtcctgta 1501 tggcccccgactggacgagagggattgtccgggaaactggacgtggccagaaaattccca 1561 gcagactccaatgtgccaggcttgggggaacccattgcccgagctcaagtgtctaaagga 1621 tggcactttcccactgcccatcggggaatcagtgactgtcactcgagatcttgagggcac 1681 ctacctctgtcgggccaggagcactcaaggggaggtcacccgcaaggtgaccgtgaatgt 1741 gctctccccccggtatgagattgtcatcatcactgtggtagcagccgcagtcataatggg 1801 cactgcaggcctcagcacgtacctctataaccgccagcggaagatcaagaaatacagact 1861 acaacaggcccaaaaagggacccccatgaaaccgaacacacaagccacgcctccctgaac 1921 ctatcccgggacagggcctcttcctcggccttcccatattggtggcagtggtgccacact 1981 gaacagagtggaagacatatgccatgcagctacacctaccggccctgggacgccggagga 2041 cagggcattgtcctcagtcagatacaacagcatttggggccatggtacctgcacacctaa 2101 aacactaggccacgcatctgatctgtagtcacatgactaagccaagaggaaggagcaaga 2161 ctcaagacatgattgatggatgttaaagtctagcctgatgagaggggaagtggtggggga 2221 gacatagccccaccatgaggacatacaactgggaaatactgaaacttgctgcctattggg 2281 tatgctgaggccccacagacttacagaagaagtggccctccatagacatgtgtagcatca 2341 aaacacaaaggcccacacttcctgacggatgccagcttgggcactgctgtctactgaccc 2401 caacccttgatgatatgtatttattcatttgttattttaccagctatttattgagtgtct 2461 tttatgtaggctaaatgaacataggtctctggcctcacggagctcccagtcctaatcaca 2521 ttcaaggtcaccaggtacagttgtacaggttgtacactgcaggagagtgcctggcaaaaa 2581 gatcaaatggggctgggacttctcattggccaacctgcctttccccagaaggagtgattt 2641 ttctatcggcacaaaagcactatatggactggtaatggttacaggttcagagattaccca 2701 gtgaggccttattcctcccttccccccaaaactgacacctttgttagccacctccccacc 2761 cacatacatttctgccagtgttcacaatgacactcagcggtcatgtctggacatgagtgc 2821 ccagggaatatgcccaagctatgccttgtcctcttgtcctgtttgcatttcactgggagc 2881 ttgcactatgcagctccagtttcctgcagtgatcagggtcctgcaagcagtggggaaggg 2941 ggccaaggtattggaggactccctcccagctttggaagcctcatccgcgtgtgtgtgtgt 3001 gtgtatgtgtagacaagctctcgctctgtcacccaggctggagtgcagtggtgcaatcat 3061 ggttcactgcagtcttgaccttttgggctcaagtgatcctcccacctcagcctcctgagt 3121 agctgggaccataggctcacaacaccacacctggcaaatttgattttttttttttttcca 3181 gagacggggtctcgcaacattgcccagacttcctttgtgttagttaataaagctttctca 3241 actgccaaa Collagen(NCBIRef.:NM_000088.3;SEQIDNO:178) 1 tcgtcggagcagacgggagtttctcctcggggtcggagcaggaggcacgcggagtgtgag 61 gccacgcatgagcggacgctaaccccctccccagccacaaagagtctacatgtctagggt 121 ctagacatgttcagctttgtggacctccggctcctgctcctcttagcggccaccgccctc 181 ctgacgcacggccaagaggaaggccaagtcgagggccaagacgaagacatcccaccaatc 241 acctgcgtacagaacggcctcaggtaccatgaccgagacgtgtggaaacccgagccctgc 301 cggatctgcgtctgcgacaacggcaaggtgttgtgcgatgacgtgatctgtgacgagacc 361 aagaactgccccggcgccgaagtccccgagggcgagtgctgtcccgtctgccccgacggc 421 tcagagtcacccaccgaccaagaaaccaccggcgtcgagggacccaagggagacactggc 481 ccccgaggcccaaggggacccgcaggcccccctggccgagatggcatccctggacagcct 541 ggacttcccggaccccccggaccccccggacctcccggaccccctggcctcggaggaaac 601 tttgctccccagctgtcttatggctatgatgagaaatcaaccggaggaatttccgtgcct 661 ggccccatgggtccctctggtcctcgtggtctccctggcccccctggtgcacctggtccc 721 caaggcttccaaggtccccctggtgagcctggcgagcctggagcttcaggtcccatgggt 781 ccccgaggtcccccaggtccccctggaaagaatggagatgatggggaagctggaaaacct 841 ggtcgtcctggtgagcgtgggcctcctgggcctcagggtgctcgaggattgcccggaaca 901 gctggcctccctggaatgaagggacacagaggtttcagtggtttggatggtgccaaggga 961 gatgctggtcctgctggtcctaagggtgagcctggcagccctggtgaaaatggagctcct 1021 ggtcagatgggcccccgtggcctgcctggtgagagaggtcgccctggagcccctggccct 1081 gctggtgctcgtggaaatgatggtgctactggtgctgccgggccccctggtcccaccggc 1141 cccgctggtcctcctggcttccctggtgctgttggtgctaagggtgaagctggtccccaa 1201 gggccccgaggctctgaaggtccccagggtgtgcgtggtgagcctggcccccctggccct 1261 gctggtgctgctggccctgctggaaaccctggtgctgatggacagcctggtgctaaaggt 1321 gccaatggtgctcctggtattgctggtgctcctggcttccctggtgcccgaggcccctct 1381 ggaccccagggccccggcggccctcctggtcccaagggtaacagcggtgaacctggtgct 1441 cctggcagcaaaggagacactggtgctaagggagagcctggccctgttggtgttcaagga 1501 ccccctggccctgctggagaggaaggaaagcgaggagctcgaggtgaacccggacccact 1561 ggcctgcccggaccccctggcgagcgtggtggacctggtagccgtggtttccctggcgca 1621 gatggtgttgctggtcccaagggtcccgctggtgaacgtggttctcctggccctgctggc 1681 cccaaaggatctcctggtgaagctggtcgtcccggtgaagctggtctgcctggtgccaag 1741 ggtctgactggaagccctggcagccctggtcctgatggcaaaactggcccccctggtccc 1801 gccggtcaagatggtcgccccggacccccaggcccacctggtgcccgtggtcaggctggt 1861 gtgatgggattccctggacctaaaggtgctgctggagagcccggcaaggctggagagcga 1921 ggtgttcccggaccccctggcgctgtcggtcctgctggcaaagatggagaggctggagct 1981 cagggaccccctggccctgctggtcccgctggcgagagaggtgaacaaggccctgctggc 2041 tcccccggattccagggtctccctggtcctgctggtcctccaggtgaagcaggcaaacct 2101 ggtgaacagggtgttcctggagaccttggcgcccctggcccctctggagcaagaggcgag 2161 agaggtttccctggcgagcgtggtgtgcaaggtccccctggtcctgctggtccccgaggg 2221 gccaacggtgctcccggcaacgatggtgctaagggtgatgctggtgcccctggagctccc 2281 ggtagccagggcgcccctggccttcagggaatgcctggtgaacgtggtgcagctggtctt 2341 ccagggcctaagggtgacagaggtgatgctggtcccaaaggtgctgatggctctcctggc 2401 aaagatggcgtccgtggtctgactggccccattggtcctcctggccctgctggtgcccct 2461 ggtgacaagggtgaaagtggtcccagcggccctgctggtcccactggagctcgtggtgcc 2521 cccggagaccgtggtgagcctggtccccccggccctgctggctttgctggcccccctggt 2581 gctgacggccaacctggtgctaaaggcgaacctggtgatgctggtgctaaaggcgatgct 2641 ggtccccctggccctgccggacccgctggaccccctggccccattggtaatgttggtgct 2701 cctggagccaaaggtgctcgcggcagcgctggtccccctggtgctactggtttccctggt 2761 gctgctggccgagtcggtcctcctggcccctctggaaatgctggaccccctggccctcct 2821 ggtcctgctggcaaagaaggcggcaaaggtccccgtggtgagactggccctgctggacgt 2881 cctggtgaagttggtccccctggtccccctggccctgctggcgagaaaggatcccctggt 2941 gctgatggtcctgctggtgctcctggtactcccgggcctcaaggtattgctggacagcgt 3001 ggtgtggtcggcctgcctggtcagagaggagagagaggcttccctggtcttcctggcccc 3061 tctggtgaacctggcaaacaaggtccctctggagcaagtggtgaacgtggtccccctggt 3121 cccatgggcccccctggattggctggaccccctggtgaatctggacgtgagggggctcct 3181 ggtgccgaaggttcccctggacgagacggttctcctggcgccaagggtgaccgtggtgag 3241 accggccccgctggaccccctggtgctcctggtgctcctggtgcccctggccccgttggc 3301 cctgctggcaagagtggtgatcgtggtgagactggtcctgctggtcccgccggtcctgtc 3361 ggccctgttggcgcccgtggccccgccggaccccaaggcccccgtggtgacaagggtgag 3421 acaggcgaacagggcgacagaggcataaagggtcaccgtggcttctctggcctccagggt 3481 ccccctggccctcctggctctcctggtgaacaaggtccctctggagcctctggtcctgct 3541 ggtccccgaggtccccctggctctgctggtgctcctggcaaagatggactcaacggtctc 3601 cctggccccattgggccccctggtcctcgcggtcgcactggtgatgctggtcctgttggt 3661 ccccccggccctcctggacctcctggtccccctggtcctcccagcgctggtttcgacttc 3721 agcttcctgccccagccacctcaagagaaggctcacgatggtggccgctactaccgggct 3781 gatgatgccaatgtggttcgtgaccgtgacctcgaggtggacaccaccctcaagagcctg 3841 agccagcagatcgagaacatccggagcccagagggcagccgcaagaaccccgcccgcacc 3901 tgccgtgacctcaagatgtgccactctgactggaagagtggagagtactggattgacccc 3961 aaccaaggctgcaacctggatgccatcaaagtcttctgcaacatggagactggtgagacc 4021 tgcgtgtaccccactcagcccagtgtggcccagaagaactggtacatcagcaagaacccc 4081 aaggacaagaggcatgtctggttcggcgagagcatgaccgatggattccagttcgagtat 4141 ggcggccagggctccgaccctgccgatgtggccatccagctgaccttcctgcgcctgatg 4201 tccaccgaggcctcccagaacatcacctaccactgcaagaacagcgtggcctacatggac 4261 cagcagactggcaacctcaagaaggccctgctcctccagggctccaacgagatcgagatc 4321 cgcgccgagggcaacagccgcttcacctacagcgtcactgtcgatggctgcacgagtcac 4381 accggagcctggggcaagacagtgattgaatacaaaaccaccaagacctcccgcctgccc 4441 atcatcgatgtggcccccttggacgttggtgccccagaccaggaattcggcttcgacgtt 4501 ggccctgtctgcttcctgtaaactccctccatcccaacctggctccctcccacccaacca 4561 actttccccccaacccggaaacagacaagcaacccaaactgaaccccctcaaaagccaaa 4621 aaatgggagacaatttcacatggactttggaaaatatttttttcctttgcattcatctct 4681 caaacttagtttttatctttgaccaaccgaacatgaccaaaaaccaaaagtgcattcaac 4741 cttaccaaaaaaaaaaaaaaaaaaagaataaataaataactttttaaaaaaggaagcttg 4801 gtccacttgcttgaagacccatgcgggggtaagtccctttctgcccgttgggcttatgaa 4861 accccaatgctgccctttctgctcctttctccacaccccccttggggcctcccctccact 4921 ccttcccaaatctgtctccccagaagacacaggaaacaatgtattgtctgcccagcaatc 4981 aaaggcaatgctcaaacacccaagtggcccccaccctcagcccgctcctgcccgcccagc 5041 acccccaggccctgggggacctggggttctcagactgccaaagaagccttgccatctggc 5101 gctcccatggctcttgcaacatctccccttcgtttttgagggggtcatgccgggggagcc 5161 accagcccctcactgggttcggaggagagtcaggaagggccacgacaaagcagaaacatc 5221 ggatttggggaacgcgtgtcaatcccttgtgccgcagggctgggcgggagagactgttct 5281 gttccttgtgtaactgtgttgctgaaagactacctcgttcttgtcttgatgtgtcaccgg 5341 ggcaactgcctgggggcggggatgggggcagggtggaagcggctccccattttataccaa 5401 aggtgctacatctatgtgatgggtggggtggggagggaatcactggtgctatagaaattg 5461 agatgcccccccaggccagcaaatgttcctttttgttcaaagtctatttttattccttga 5521 tatttttctttttttttttttttttttgtggatggggacttgtgaatttttctaaaggtg 5581 ctatttaacatgggaggagagcgtgtgcggctccagcccagcccgctgctcactttccac 5641 cctctctccacctgcctctggcttctcaggcctctgctctccgacctctctcctctgaaa 5701 ccctcctccacagctgcagcccatcctcccggctccctcctagtctgtcctgcgtcctct 5761 gtccccgggtttcagagacaacttcccaaagcacaaagcagtttttccccctaggggtgg 5821 gaggaagcaaaagactctgtacctattttgtatgtgtataataatttgagatgtttttaa 5881 ttattttgattgctggaataaagcatgtggaaatgacccaaacataa E-cadherin(NCBIRef.:NM_001317184.1;SEQIDNO:179) 1 tcagtggcgtcggaactgcaaagcacctgtgagcttgcggaagtcagttcagactccagc 61 ccgctccagcccggcccgacccgaccgcacccggcgcctgccctcgctcggcgtccccgg 121 ccagccatgggcccttggagccgcagcctctcggcgctgctgctgctgctgcaggtctcc 181 tcttggctctgccaggagccggagccctgccaccctggctttgacgccgagagctacacg 241 ttcacggtgccccggcgccacctggagagaggccgcgtcctgggcagagtgaattttgaa 301 gattgcaccggtcgacaaaggacagcctatttttccctcgacacccgattcaaagtgggc 361 acagatggtgtgattacagtcaaaaggcctctacggtttcataacccacagatccatttc 421 ttggtctacgcctgggactccacctacagaaagttttccaccaaagtcacgctgaataca 481 gtggggcaccaccaccgccccccgccccatcaggcctccgtttctggaatccaagcagaa 541 ttgctcacatttcccaactcctctcctggcctcagaagacagaagagagactgggttatt 601 cctcccatcagctgcccagaaaatgaaaaaggcccatttcctaaaaacctggttcagatc 661 aaatccaacaaagacaaagaaggcaaggttttctacagcatcactggccaaggagctgac 721 acaccccctgttggtgtctttattattgaaagagaaacaggatggctgaaggtgacagag 781 cctctggatagagaacgcattgccacatacactctcttctctcacgctgtgtcatccaac 841 gggaatgcagttgaggatccaatggagattttgatcacggtaaccgatcagaatgacaac 901 aagcccgaattcacccaggaggtctttaaggggtctgtcatggaaggtgctcttccagga 961 acctctgtgatggaggtcacagccacagacgcggacgatgatgtgaacacctacaatgcc 1021 gccatcgcttacaccatcctcagccaagatcctgagctccctgacaaaaatatgttcacc 1081 attaacaggaacacaggagtcatcagtgtggtcaccactgggctggaccgagagagtttc 1141 cctacgtataccctggtggttcaagctgctgaccttcaaggtgaggggttaagcacaaca 1201 gcaacagctgtgatcacagtcactgacaccaacgataatcctccgatcttcaatcccacc 1261 acgggcttggattttgaggccaagcagcagtacattctacacgtagcagtgacgaatgtg 1321 gtaccttttgaggtctctctcaccacctccacagccaccgtcaccgtggatgtgctggat 1381 gtgaatgaagcccccatctttgtgcctcctgaaaagagagtggaagtgtccgaggacttt 1441 ggcgtgggccaggaaatcacatcctacactgcccaggagccagacacatttatggaacag 1501 aaaataacatatcggatttggagagacactgccaactggctggagattaatccggacact 1561 ggtgccatttccactcgggctgagctggacagggaggattttgagcacgtgaagaacagc 1621 acgtacacagccctaatcatagctacagacaatggttctccagttgctactggaacaggg 1681 acacttctgctgatcctgtctgatgtgaatgacaacgcccccataccagaacctcgaact 1741 atattcttctgtgagaggaatccaaagcctcaggtcataaacatcattgatgcagacctt 1801 cctcccaatacatctcccttcacagcagaactaacacacggggcgagtgccaactggacc 1861 attcagtacaacgacccaacccaagaatctatcattttgaagccaaagatggccttagag 1921 gtgggtgactacaaaatcaatctcaagctcatggataaccagaataaagaccaagtgacc 1981 accttagaggtcagcgtgtgtgactgtgaaggggccgctggcgtctgtaggaaggcacag 2041 cctgtcgaagcaggattgcaaattcctgccattctggggattcttggaggaattcttgct 2101 ttgctaattctgattctgctgctcttgctgtttcttcggaggagagcggtggtcaaagag 2161 cccttactgcccccagaggatgacacccgggacaacgtttattactatgatgaagaagga 2221 ggcggagaagaggaccaggactttgacttgagccagctgcacaggggcctggacgctcgg 2281 cctgaagtgactcgtaacgacgttgcaccaaccctcatgagtgtcccccggtatcttccc 2341 cgccctgccaatcccgatgaaattggaaattttattgatgaaaatctgaaagcggctgat 2401 actgaccccacagccccgccttatgattctctgctcgtgtttgactatgaaggaagcggt 2461 tccgaagctgctagtctgagctccctgaactcctcagagtcagacaaagaccaggactat 2521 gactacttgaacgaatggggcaatcgcttcaagaagctggctgacatgtacggaggcggc 2581 gaggacgactaggggactcgagagaggcgggccccagacccatgtgctgggaaatgcaga 2641 aatcacgttgctggtggtttttcagctcccttcccttgagatgagtttctggggaaaaaa 2701 aagagactggttagtgatgcagttagtatagctttatactctctccactttatagctcta 2761 ataagtttgtgttagaaaagtttcgacttatttcttaaagcttttttttttttcccatca 2821 ctctttacatggtggtgatgtccaaaagatacccaaattttaatattccagaagaacaac 2881 tttagcatcagaaggttcacccagcaccttgcagattttcttaaggaattttgtctcact 2941 tttaaaaagaaggggagaagtcagctactctagttctgttgttttgtgtatataattttt 3001 taaaaaaaatttgtgtgcttctgctcattactacactggtgtgtccctctgccttttttt 3061 tttttttaagacagggtctcattctatcggccaggctggagtgcagtggtgcaatcacag 3121 ctcactgcagccttgtcctcccaggctcaagctatccttgcacctcagcctcccaagtag 3181 ctgggaccacaggcatgcaccactacgcatgactaattttttaaatatttgagacggggt 3241 ctccctgtgttacccaggctggtctcaaactcctgggctcaagtgatcctcccatcttgg 3301 cctcccagagtattgggattacagacatgagccactgcacctgcccagctccccaactcc 3361 ctgccattttttaagagacagtttcgctccatcgcccaggcctgggatgcagtgatgtga 3421 tcatagctcactgtaacctcaaactctggggctcaagcagttctcccaccagcctccttt 3481 ttatttttttgtacagatggggtcttgctatgttgcccaagctggtcttaaactcctggc 3541 ctcaagcaatccttctgccttggccccccaaagtgctgggattgtgggcatgagctgctg 3601 tgcccagcctccatgttttaatatcaactctcactcctgaattcagttgctttgcccaag 3661 ataggagttctctgatgcagaaattattgggctcttttagggtaagaagtttgtgtcttt 3721 gtctggccacatcttgactaggtattgtctactctgaagacctttaatggcttccctctt 3781 tcatctcctgagtatgtaacttgcaatgggcagctatccagtgacttgttctgagtaagt 3841 gtgttcattaatgtttatttagctctgaagcaagagtgatatactccaggacttagaata 3901 gtgcctaaagtgctgcagccaaagacagagcggaactatgaaaagtgggcttggagatgg 3961 caggagagcttgtcattgagcctggcaatttagcaaactgatgctgaggatgattgaggt 4021 gggtctacctcatctctgaaaattctggaaggaatggaggagtctcaacatgtgtttctg 4081 acacaagatccgtggtttgtactcaaagcccagaatccccaagtgcctgcttttgatgat 4141 gtctacagaaaatgctggctgagctgaacacatttgcccaattccaggtgtgcacagaaa 4201 accgagaatattcaaaattccaaatttttttcttaggagcaagaagaaaatgtggcccta 4261 aagggggttagttgaggggtagggggtagtgaggatcttgatttggatctctttttattt 4321 aaatgtgaatttcaacttttgacaatcaaagaaaagacttttgttgaaatagctttactg 4381 tttctcaagtgttttggagaaaaaaatcaaccctgcaatcactttttggaattgtcttga 4441 tttttcggcagttcaagctatatcgaatatagttctgtgtagagaatgtcactgtagttt 4501 tgagtgtatacatgtgtgggtgctgataattgtgtattttctttgggggtggaaaaggaa 4561 aacaattcaagctgagaaaagtattctcaaagatgcatttttataaattttattaaacaa 4621 ttttgttaaaccattaaaaaaaaaaaaaaaaaaaaaaaaaaa Laminin(LAMA1)(NCBIRef.:NM_005559.3;SEQIDNO:180) 1 cggggccagggcagcgcggactcgcgtcccgtggagcgttccaggcgggcgcgcggcttt 61 ctccccagacccaccgagtggcggcggaggcgagatgcgcgggggcgtgctcctggtctt 121 gctgctgtgtgtcgccgcgcagtgccggcagagaggcctgtttcctgccattctcaatct 181 tgccagcaatgctcacatcagcaccaatgccacctgtggcgagaaggggccggagatgtt 241 ctgcaaacttgtggagcatgtgccaggtcggcccgtccgaaacccacagtgccggatctg 301 tgatggcaacagcgcaaaccccagagaacgccatccaatatcacatgccatagatggcac 361 caataactggtggcaaagtcccagcattcagaatgggagagaatatcactgggtcacaat 421 cactctggacttaagacaggtctttcaagttgcatatgtcatcattaaagctgccaatgc 481 ccctcgacctggaaactggattttggagcgttctctggatggcaccacgttcagcccctg 541 gcagtattatgcagtcagcgactcagagtgtttgtctcgttacaatataactccaagacg 601 agggccacccacctacagggctgatgatgaagtgatctgcacctcctattattccagatt 661 ggtgccacttgagcatggagagattcatacatcactcatcaatggcagaccaagcgctga 721 cgatctttcacccaagttgttggaattcacttctgcacgatatattcgccttcgcttgca 781 acgcattagaacgctcaatgcagatctcatgacccttagccaccgggaacctaaagaact 841 ggatcctattgttaccagacgctattattattcaataaaggacatttctgttggaggcat 901 gtgtatctgctatggccatgctagtagctgcccatgggatgaaactacaaagaaactgca 961 gtgtcaatgtgagcataatacttgcggggagagctgtaacaggtgctgtcctgggtacca 1021 tcagcagccctggaggccgggaaccgtgtcctccggcaatacatgtgaagcatgtaattg 1081 tcacaataaagccaaagactgttactatgatgaaagtgttgcaaagcagaagaaaagttt 1141 gaatactgctggacagttcagaggaggaggggtttgcataaattgcttgcagaacaccat 1201 gggaatcaactgtgaaacctgtattgatggatattatagaccacacaaagtgtctcctta 1261 tgaggatgagccttgccgcccctgtaattgtgaccctgtggggtccctcagttctgtctg 1321 tattaaggatgacctccattctgacttacacaatgggaagcagccaggtcagtgcccatg 1381 taaggaaggttatacaggagaaaaatgtgatcgctgccaacttggctataaggattaccc 1441 gacctgtgtctcctgtgggtgcaacccagtgggcagtgccagtgatgagccctgcacagg 1501 gccctgtgtttgtaaggaaaacgttgaggggaaggcctgtgatcgctgcaagccaggatt 1561 ctataacttgaaggaaaaaaacccccggggctgctccgagtgcttctgctttggcgtttc 1621 tgatgtctgcagcagcctctcttggcctgttggtcaggtaaacagtatgtccgggtggct 1681 ggtcaccgacttgatcagtcccaggaagatcccgtctcagcaagatgcactaggcgggcg 1741 ccatcaggtcagcatcaacaacaccgcggtcatgcagagactggctcccaagtactactg 1801 ggcagcccccgaggcctaccttggaaataagctgactgcgtttggcggattcctgaaata 1861 cacggtgtcctacgatattccggtagagacggtagacagtaacctcatgtcgcatgctga 1921 cgtcatcattaagggaaacggactcactttaagcacacaggctgagggtctgtcattgca 1981 gccttatgaagagtacctaaacgtggttagacttgtgcctgaaaacttccaagattttca 2041 cagcaaaaggcagattgatcgtgaccagctgatgactgtccttgccaatgtgacacatct 2101 tttgatcagagccaactacaattctgcaaaaatggctctttacaggttggagtccgtctc 2161 tctggacatagccagctctaatgccatcgacctggtggtggccgctgatgtggagcactg 2221 tgaatgtccgcaaggctacacagggacctcctgtgagtcgtgcctctctggctattaccg 2281 cgtggatggaatactctttggaggaatttgtcaaccctgtgaatgccacggccatgcagc 2341 tgagtgtaatgttcacggcgtttgcattgcgtgtgcgcacaacaccaccggcgtccactg 2401 tgagcagtgcttgcccggcttctacggggagccttcccgagggacacctggggactgcca 2461 gccctgcgcctgccctctcaccatagcctccaacaatttcagccccacctgccacctcaa 2521 tgatggagatgaagtggtctgtgactggtgtgccccgggctactcaggagcttggtgtga 2581 gagatgtgcagatggttactatggaaacccaacagtgcctggcgaatcttgtgttccctg 2641 tgactgcagcggcaacgtggacccctcggaggctggtcactgtgactcagtcaccgggga 2701 gtgcctgaagtgcctggggaacacagatggcgcccactgtgaaaggtgtgctgacgggtt 2761 ctatggggacgctgtgacagccaagaactgccgcgcctgtgaatgccatgtgaaaggctc 2821 ccattctgccgtgtgccatcttgagaccgggctctgtgactgcaaaccaaacgtgactgg 2881 acagcagtgtgaccagtgcttgcatggctattatgggctggactcaggccatggctgccg 2941 gccctgcaactgcagcgtggcaggctccgtgtcagatggctgcacggatgaaggccagtg 3001 tcactgtgtcccaggtgtggcagggaaaaggtgtgacaggtgtgcccatggcttctacgc 3061 ctaccaggatggtagctgtacaccctgtgactgcccacacactcagaatacctgcgaccc 3121 agaaactggagagtgtgtctgcccccctcacacacagggtgtgaagtgtgaagaatgtga 3181 ggatgggcactggggctacgatgcggaggtggggtgccaggcctgcaattgcagtctcgt 3241 ggggtcgactcatcatcggtgcgatgtggtcaccggccattgccagtgcaagtcaaaatt 3301 tggtggccgggcctgcgatcagtgttccttgggttacagagactttcccgactgtgttcc 3361 ctgtgactgtgacctgagggggacgtcgggggacgcctgcaacctggagcagggtctctg 3421 cggctgtgtggaggaaaccggggcctgcccttgcaaggaaaatgtctttggtcctcagtg 3481 caacgaatgtcgagagggcaccttcgctctccgcgcagacaaccccctgggctgcagccc 3541 gtgcttctgctccgggctgtcccacctctgctcagagctggaggactacgtgaggacccc 3601 agtaacgctgggctccgatcagcctcttctgcgtgtggtttctcagagtaacttgagggg 3661 cacgaccgagggggtttactaccaggcccccgacttcctgctggatgccgccaccgtccg 3721 gcagcacatccgtgcagagccgttttactggcggctgccgcagcagttccaaggagacca 3781 gctcatggcctatggtggcaaactgaagtacagcgtggccttctattctttggatggcgt 3841 cggcacctccaattttgagcctcaagttctcatcaaaggtggtcggatcagaaagcaagt 3901 catttacatggatgcaccagccccagagaatggagtgagacaggaacaagaagtagcaat 3961 gagagagaatttttggaaatattttaactctgtttctgaaaaacctgtcacgcgagagga 4021 ttttatgtctgtcctcagcgatattgagtacatcctcatcaaggcatcgtatggtcaagg 4081 attacagcagagcagaatctcagacatttcaatggaggttggcagaaaggctgaaaagct 4141 gcacccagaagaagaggttgcatctcttttagagaattgtgtctgtcctcctggcactgt 4201 gggattctcatgtcaggactgcgcccctgggtaccacagagggaagctcccagcagggag 4261 tgacaggggaccacgccctctggttgctccttgtgttccctgcagttgcaacaaccacag 4321 tgacacctgtgaccccaacaccgggaagtgtctgaactgtggcgataacacagcaggtga 4381 ccattgtgatgtgtgtacttctggctactacgggaaggtgactggctcagcaagtgactg 4441 tgctctgtgtgcctgtcctcacagccctcctgccagttttagtcccacttgtgtcttgga 4501 aggggaccacgatttccgttgtgacgcctgtctcctgggctatgaaggaaaacactgtga 4561 aaggtgctcctcaagctattatgggaaccctcaaacaccaggtggcagttgccagaagtg 4621 tgactgcaacccgcacggctctgtccacggtgactgtgaccgcacatctgggcagtgcgt 4681 ttgcaggctgggggcctcggggctccggtgcgatgagtgtgaaccgaggcacattctgat 4741 ggaaacagattgtgtttcctgtgatgatgagtgtgtaggtgtgctgctgaatgacttgga 4801 tgagattggtgatgccgttctttctctgaacctcactggcattatccctgtcccatatgg 4861 aattttgtcaaacctggaaaatacaactaaatatctccaggaatctttattaaaagaaaa 4921 tatgcaaaaggacctgggaaaaattaagcttgaaggtgttgcagaagaaacggacaacct 4981 gcaaaagaagctcactaggatgttagcgagtacccaaaaggtgaatagggcaactgagag 5041 aatcttcaaggagagtcaagacctggccatagccattgagaggctgcagatgagcatcac 5101 agaaattatggaaaagacaactttaaatcagactttggatgaagatttcctactacccaa 5161 ttctactcttcagaacatgcaacagaatggtacatctttgctagaaatcatgcagataag 5221 agacttcacacagttgcaccaaaatgccacccttgaactcaaggctgctgaagatttatt 5281 gtcacaaattcaggaaaattaccagaagccgctggaagaattggaggtattgaaagaagc 5341 agcaagccacgtcctttcaaagcacaacaatgaactaaaggcggctgaggcgctcgtgag 5401 ggaagctgaggcaaagatgcaggaaagcaaccacctgctgctcatggtcaatgctaatct 5461 gagagaattcagtgataaaaagctgcatgttcaagaagaacaaaatctgacctcagagct 5521 cattgtccaaggaagaggattgatagatgctgctgctgcacaaacagatgctgtacaaga 5581 tgctctagagcacttagaggatcaccaggataagctacttttatggtctgccaaaatcag 5641 gcaccacatagatgacctggtcatgcacatgtcccaaaggaacgcagtcgacctggtcta 5701 cagagctgaggaccatgccgctgagttccagagactagcagatgttctgtacagtggcct 5761 tgaaaacatcagaaatgtgtccctgaatgccaccagtgcagcctatgtccattacaacat 5821 ccagagcctgattgaagaatcggaggaactggccagagatgctcacaggactgtgactga 5881 gacgagcctgctctcagaatcccttgtttctaacgggaaagcggccgtgcagcgcagctc 5941 cagatttctaaaagaaggcaacaacctcagcaggaagcttccaggtattgcattggaact 6001 gagtgaattgagaaataagacaaacagatttcaagagaatgctgttgaaattaccaggca 6061 aaccaatgaatcactcttgatacttagagcaattcctaaaggtataagagacaagggagc 6121 caaaaccaaagagctggccacgtctgcaagccagagcgcggtgagcacgctgagggacgt 6181 ggcggggctgagccaggagctgctgaacacatctgccagcctgtccagggtcaacaccac 6241 attacgagagacacaccagcttctgcaggactccaccatggccactctgttggctggaag 6301 aaaagtcaaagacgtggaaattcaagccaaccttttgtttgatcggttgaagcctttgaa 6361 gatgttagaggagaatctgagcagaaacctatcagaaattaaactgttgatcagccaggc 6421 ccgcaaacaagcagcttctattaaagtcgccgtgtctgcagacagagattgcatccgggc 6481 ctaccagcctcagatttcctctaccaactacaataccttaacactaaatgttaagacaca 6541 ggaacccgataatcttctcttctacctcggtagcagcaccgcttctgatttccttgcagt 6601 ggagatgcggcgagggagagtggccttcctgtgggacctgggctccgggtccacacgctt 6661 ggagtttccagactttcccattgatgacaacagatggcacagtatccatgtagccagatt 6721 tggaaacattggttcactgagtgtaaaggaaatgagctcaaatcaaaagtcaccaacaaa 6781 aacaagtaaatcccctgggacagctaatgttctggatgtaaacaattcaacactcatgtt 6841 tgttggaggtcttggaggacaaatcaagaaatctcctgctgtgaaggttactcattttaa 6901 aggctgcttgggggaggccttcctgaatggaaaatccataggcctatggaactatattga 6961 aagggaaggcaagtgccgtgggtgcttcggaagctcccagaatgaagacccttccttcca 7021 ttttgacgggagtgggtactctgtcgtggagaagtcacttccggctaccgtgacccagat 7081 aatcatgctttttaataccttttcacctaatggacttcttctctacctgggttcatacgg 7141 cacaaaagactttttatccatcgagctgtttcgtggcagagtgaaggttatgactgacct 7201 gggttcaggacccattacccttttgacagacagacgttataacaatggaacctggtacaa 7261 aattgccttccagcgaaaccggaagcaaggagtgctagcagttatcgatgcctataacac 7321 cagtaataaagaaaccaagcagggcgagactccgggagcatcttctgacctcaaccgcct 7381 agacaaggacccgatttatgtgggtggattaccaaggtcaagagttgtaaggagaggtgt 7441 caccaccaaaagctttgtgggctgcatcaagaacctggaaatatccagatcaacctttga 7501 cttactcagaaattcctatggagtgagaaaaggctgtttactggagcccatccggagtgt 7561 tagcttcctgaaaggcggctacattgaattgccacccaaatctttgtcaccagaatcaga 7621 atggctggtaacatttgccaccacgaacagcagtggcatcatcctggctgccctcggcgg 7681 ggatgtggagaagcggggtgatcgtgaggaagcacacgtgcccttcttttccgtcatgct 7741 gatcggaggcaacattgaggtacatgtcaatcctggggatgggacaggcctgagaaaagc 7801 tctcctgcacgctcccacgggtacctgcagtgatggacaagcgcattccatctccttggt 7861 caggaatcggagaattatcactgtccaattggatgagaacaatcctgtggaaatgaagtt 7921 gggcacattagtagaaagcaggacgataaatgtgtccaatctgtacgtcgggggaattcc 7981 agagggagaggggacgtcactgctcacaatgagaagatcgttccatggctgtatcaaaaa 8041 cctgatcttcaatttggaacttttggatttcaacagtgcagttggccatgagcaagtcga 8101 cctggacacctgctggctgtcagaaaggcctaagctggctcccgatgcagaggacagcaa 8161 gctcttgccagagccccgggcttttccagaacagtgtgtggtggatgcagctctggagta 8221 cgttcccggcgctcaccagtttggtctcacacaaaacagccatttcatcttgccttttaa 8281 tcagtcggctgtcagaaagaagctctcggttgagctaagcatccgcacgttcgcctccag 8341 cggcctgatttactacatggctcatcagaaccaagcagactacgctgtgctccagctgca 8401 cgggggccgcctccacttcatgtttgaccttggcaaaggcagaacaaaggtctctcaccc 8461 tgcactgctcagtgatggcaagtggcacacggtcaagacagactatgttaaaagaaaagg 8521 cttcataactgtcgacggccgagagtctcccatggtgactgtggtgggagatggaaccat 8581 gctggatgtggagggtttgttctacctaggaggcctgccctcccagtaccaggccaggaa 8641 aattggaaatatcacccacagcatccctgcctgcattggggatgtgacggttaacagcaa 8701 acagctggacaaggacagcccggtgtctgccttcacggtgaacaggtgctacgcagtggc 8761 ccaggaaggaacatactttgacggaagcggatatgcagctcttgtcaaagagggctacaa 8821 agtccagtcagatgtgaacatcacactggagtttcgaacctcctcgcagaatggcgtcct 8881 cctggggatcagcactgccaaagtggatgccattggactagagcttgtggacggcaaggt 8941 cttgttccatgtcaacaatggtgctggcaggataacagctgcatatgagcccaaaaccgc 9001 cactgtgctctgtgatggaaaatggcacactcttcaagctaacaaaagcaaacaccgtat 9061 cactctgattgttgacgggaacgcagttggcgctgaaagtccacacacccagtctacctc 9121 agtggacaccaacaatcccatttatgttggtggctatcctgctggtgtgaagcaaaaatg 9181 cctgcgcagccagacctcgttccgcgggtgtttgaggaagctagctctgattaagagccc 9241 gcaggtgcagtcctttgacttcagcagagcgttcgaactgcacggagttttccttcattc 9301 ctgtcctgggaccgagtcctgaacttcaagcagaatcctcagttggaatcattgctaata 9361 ttttgaggagaagtgtatgtgtgaattaagaatctcttcagttcatatttcatttccaac 9421 tcaggttaagtgtttctggggagagatgttgtgtttacgttacactaaaaccacatgtgc 9481 aacaaatacctccattaaatggtctaaaatgtaaattgaattccctggctctctttttaa 9541 acgtatttttaaaaaaatctttatacacattgaatgttctgttgattacttgatagtatt 9601 ttatgtttttcattttgagctttttaaaaaagtatcaatacagatgataacagatca Fibulin-5(NCBIRef.:NM_006329.3;SEQIDNO:181) 1 cgcccctcgccttctgcccgggcgctcgcagccgagcgcggccggggaagggctctcctc 61 ccagcgccgagcactgggccctggcagacgccccaagattgttgtgaggagtctagccag 121 ttggtgagcgctgtaatctgaaccagctgtgtccagactgaggccccatttgcattgttt 181 aacatacttagaaaatgaagtgttcatttttaacattcctcctccaattggtttaatgct 241 gaattactgaagagggctaagcaaaaccaggtgcttgcgctgagggctctgcagtggctg 301 ggaggaccccggcgctctccccgtgtcctctccacgactcgctcggcccctctggaataa 361 aacacccgcgagccccgagggcccagaggaggccgacgtgcccgagctcctccgggggtc 421 ccgcccgcgagctttcttctcgccttcgcatctcctcctcgcgcgtcttggacatgccag 481 gaataaaaaggatactcactgttaccattctggctctctgtcttccaagccctgggaatg 541 cacaggcacagtgcacgaatggctttgacctggatcgccagtcaggacagtgtttagata 601 ttgatgaatgccgaaccatccccgaggcctgccgaggagacatgatgtgtgttaaccaaa 661 atggcgggtatttatgcattccccggacaaaccctgtgtatcgagggccctactcgaacc 721 cctactcgaccccctactcaggtccgtacccagcagctgccccaccactctcagctccaa 781 actatcccacgatctccaggcctcttatatgccgctttggataccagatggatgaaagca 841 accaatgtgtggatgtggacgagtgtgcaacagattcccaccagtgcaaccccacccaga 901 tctgcatcaatactgaaggcgggtacacctgctcctgcaccgacggatattggcttctgg 961 aaggccagtgcttagacattgatgaatgtcgctatggttactgccagcagctctgtgcga 1021 atgttcctggatcctattcttgtacatgcaaccctggttttaccctcaatgaggatggaa 1081 ggtcttgccaagatgtgaacgagtgtgccaccgagaacccctgcgtgcaaacctgcgtca 1141 acacctacggctctttcatctgccgctgtgacccaggatatgaacttgaggaagatggcg 1201 ttcattgcagtgatatggacgagtgcagcttctctgagttcctctgccaacatgagtgtg 1261 tgaaccagcccggcacatacttctgctcctgccctccaggctacatcctgctggatgaca 1321 accgaagctgccaagacatcaacgaatgtgagcacaggaaccacacgtgcaacctgcagc 1381 agacgtgctacaatttacaagggggcttcaaatgcattgaccccatccgctgtgaggagc 1441 cttatctgaggatcagtgataaccgctgtatgtgtcctgctgagaaccctggctgcagag 1501 accagccctttaccatcttgtaccgggacatggacgtggtgtcaggacgctccgttcccg 1561 ctgacatcttccaaatgcaagccacgacccgctaccctggggcctattacattttccaga 1621 tcaaatctgggaatgagggcagagaattttacatgcggcaaacgggccccatcagtgcca 1681 ccctggtgatgacacgccccatcaaagggccccgggaaatccagctggacttggaaatga 1741 tcactgtcaacactgtcatcaacttcagaggcagctccgtgatccgactgcggatatatg 1801 tgtcgcagtacccattctgagcctcgggctggagcctccgacgctgcctctcattggcac 1861 caagggacaggagaagagaggaaataacagagagaatgagagcgacacagacgttaggca 1921 tttcctgctgaacgtttccccgaagagtcagccccgacttcctgactctcacctgtacta 1981 ttgcagacctgtcaccctgcaggacttgccacccccagttcctatgacacagttatcaaa 2041 aagtattatcattgctcccctgatagaagattgttggtgaattttcaaggccttcagttt 2101 atttccactattttcaaagaaaatagattaggtttgcgggggtctgagtctatgttcaaa 2161 gactgtgaacagcttgctgtcacttcttcacctcttccactccttctctcactgtgttac 2221 tgctttgcaaagacccgggagctggcggggaaccctgggagtagctagtttgctttttgc 2281 gtacacagagaaggctatgtaaacaaaccacagcaggatcgaagggtttttagagaatgt 2341 gtttcaaaaccatgcctggtattttcaaccataaaagaagtttcagttgtccttaaattt 2401 gtataacggtttaattctgtcttgttcattttgagtatttttaaaaaatatgtcgtagaa 2461 ttccttcgaaaggccttcagacacatgctatgttctgtcttcccaaacccagtctcctct 2521 ccattttagcccagtgttttctttgaggaccccttaatcttgctttctttagaattttta 2581 cccaattggattggaatgcagaggtctccaaactgattaaatatttgaagagaaaaa

[0775] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a target integrin or a target integrin ligand (e.g., any of the exemplary target integrins or any of the exemplary integrin ligands described herein). Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0776] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a target integrin (e.g., any of the exemplary target integrins described herein) or a nucleic acid encoding an integrin ligands (e.g., any of the exemplary integrin ligands described herein). Antisense nucleic acids targeting a nucleic acid encoding a target integrin (e.g., any of the exemplary integrins described herein) or a nucleic acid encoding an integrin ligand (e.g., any of the exemplary integrin ligands described herein) can be designed using the software available at the Integrated DNA Technologies website.

[0777] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0778] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0779] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a target integrin (e.g., any of the exemplary target integrins described herein) or encoding a integrin ligand (e.g., any of the exemplary integrin ligands described herein) to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0780] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 0-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0781] Exemplary integrin inhibitors that are antisense nucleic acids include ATL1102 (e.g., Limmroth et al., Neurology 83(20):1780-1788, 2014; Li et al., Dig. Liver Dis. 39(6):557-565, 2007; Goto et al., Inflamm. Bowel Dis. 12(8):758-765, 2006).

[0782] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) can be designed based upon the nucleotide sequence of any of the integrin mRNA sequences or integrin ligand mRNA sequences disclosed herein or known in the art. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a target integrin mRNA or an integrin ligand mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, an integrin mRNA (e.g., any of the exemplary integrin mRNAs described herein) or an integrin ligand mRNA (e.g., any of the exemplary integrin ligand mRNAs described herein) can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0783] An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the target integrin (e.g., any of the exemplary target integrins described herein) or the integrin ligand (e.g., any of the exemplary integrin ligands described herein) (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0784] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Nat. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0785] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0786] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0787] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Nat. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Nat. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res., 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0788] Another means by which expression of a target integrin (e.g., any of the exemplary target integrins described herein) mRNA or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein)) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0789] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Nat. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0790] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0791] The siRNA molecules used to decrease expression of a target integrin (e.g., any of the exemplary target integrins described herein) mRNA or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0792] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing a target integrin (e.g., any of the exemplary target integrins described herein) mRNA or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 132-158, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0793] As described herein, inhibitory nucleic acids preferentially bind (e.g., hybridize) to a nucleic acid encoding a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein).

[0794] Non-limiting examples of integrin inhibitors that are short interfering RNAs (siRNAs) are described in Wang et al., Cancer Cell Int. 16:90, 2016). In some embodiments, the integrin inhibitor is a short hairpin RNA (shRNA).

[0795] Non-limiting examples of integrin inhibitors that are microRNA include miR-124 (Cai et al., Sci. Rep. 7:40733, 2017), miR-134 (Qin et al., Oncol. Rep. 37(2):823-830, 2017), miR-92b (Ma et al., Oncotarget 8(4):6681-6690, 2007), miR-17 (Gong et al., Oncol. Rep. 36(4), 2016), miR-338 (Chen et al., Oncol. Rep. 36(3):1467-74, 2016), and miR-30a-5p (Li et al., Int. J. Oncol. 48(3):1155-1164, 2016).

[0796] In some embodiments, the integrin inhibitor can include modified bases/locked nucleic acids (LNAs). In some embodiments, the integrin inhibitor is an aptamer (e.g., Berg et al., Mol. Ther. Nucl. Acids 5:e294, 2016; and Hussain et al., Nucleic Acid Ther. 23(3):203-212, 2013). Additional examples of integrin inhibitors that are inhibitory nucleic acids are described in Juliano et al., Theranostics 1:211-219, 2011; Millard et al., Theranostics 1:154-188, 2011; and Teoh et al., Curr. Mol. Med. 15:714-734, 2015. In some embodiments, the integrin inhibitor is an antisense nucleic acid, e.g., alicaforsen (Yacyshyn et al., Clin. Gastroenterol. Hepatol. 5(2):215-220, 2007).

[0797] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a target integrin (e.g., any of the exemplary target integrins described herein) or an integrin ligand (e.g., any of the exemplary integrin ligands described herein) can be administered to a subject (e.g., a human subject) in need thereof.

[0798] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0799] As is known in the art, the term thermal melting point (Tm) refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

[0800] In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding a target integrin, e.g., any of the exemplary target integrins described herein, or a nucleic acid encoding an integrin ligand, e.g., any of the exemplary integrin ligands described herein) with a T.sub.m of greater than 20 C., greater than 22 C., greater than 24 C., greater than 26 C., greater than 28 C., greater than 30 C., greater than 32 C., greater than 34 C., greater than 36 C., greater than 38 C., greater than 40 C., greater than 42 C., greater than 44 C., greater than 46 C., greater than 48 C., greater than 50 C., greater than 52 C., greater than 54 C., greater than 56 C., greater than 58 C., greater than 60 C., greater than 62 C., greater than 64 C., greater than 66 C., greater than 68 C., greater than 70 C., greater than 72 C., greater than 74 C., greater than 76 C., greater than 78 C., or greater than 80 C., e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

[0801] In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding a target integrin, e.g., any of the exemplary target integrins described herein, or a nucleic acid encoding an integrin ligand, e.g., any of the exemplary integrin ligands described herein) with a T.sub.m of about 20 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., about 24 C., or about 22 C. (inclusive); about 22 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., about 26 C., or about 24 C. (inclusive); about 24 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., about 28 C., or about 26 C. (inclusive); about 26 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., about 30 C., or about 28 C. (inclusive); about 28 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., about 32 C., or about 30 C. (inclusive); about 30 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., about 34 C., or about 32 C. (inclusive); about 32 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., about 36 C., or about 34 C. (inclusive); about 34 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., about 38 C., or about 36 C. (inclusive); about 36 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., about 40 C., or about 38 C. (inclusive); about 38 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., about 42 C., or about 40 C. (inclusive); about 40 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., about 44 C., or about 42 C. (inclusive); about 42 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., about 46 C., or about 44 C. (inclusive); about 44 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., about 48 C., or about 46 C. (inclusive); about 46 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., about 50 C., or about 48 C. (inclusive); about 48 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., about 52 C., or about 50 C. (inclusive); about 50 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., about 54 C., or about 52 C. (inclusive); about 52 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., about 56 C., or about 54 C. (inclusive); about 54 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., about 58 C., or about 56 C. (inclusive); about 56 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., about 60 C., or about 58 C. (inclusive); about 58 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., about 62 C., or about 60 C. (inclusive); about 60 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., about 64 C., or about 62 C. (inclusive); about 62 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., about 66 C., or about 64 C. (inclusive); about 64 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., about 68 C., or about 66 C. (inclusive); about 66 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., about 70 C., or about 68 C. (inclusive); about 68 C. to about 80 C., about 78 C., about 76 C., about 74 C., about 72 C., or about 70 C. (inclusive); about 70 C. to about 80 C., about 78 C., about 76 C., about 74 C., or about 72 C. (inclusive); about 72 C. to about 80 C., about 78 C., about 76 C., or about 74 C. (inclusive); about 74 C. to about 80 C., about 78 C., or about 76 C. (inclusive); about 76 C. to about 80 C. or about 78 C. (inclusive); or about 78 C. to about 80 C. (inclusive),

[0802] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater. Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0803] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Opin. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi: 10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US 2016/0090598.

[0804] In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.

[0805] In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0806] Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0807] Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.

[0808] Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.

[0809] In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

[0810] In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds. In some examples, certain organic solvents such as dimethylsulfoxide are used.

[0811] In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.

[0812] In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0813] In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.

[0814] In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

[0815] In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting an integrin can be administered to a subject (e.g., a human subject) in need of thereof.

[0816] In certain embodiments, the inhibitory nucleic acids are 10 to 40 (e.g., 10 to 30, 10 to 25, 10 to 20, 10 to 15, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40) nucleotides in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of the DNA or RNA.

Antibodies

[0817] In some embodiments, the integrin inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv).sub.2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv.sub.2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab).sub.2-scFV.sub.2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0818] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0819] Any of the antibodies or antigen-binding fragments thereof described herein can bind to any of the integrins described herein or any of the integrin ligands described herein.

[0820] In some embodiments, the antibody is a pan-31 antibody (e.g., OS2966 (Carbonell et al., Cancer Res. 73(10):3145-3154, 2013). In some embodiments, the integrin antibody is a monoclonal antibody (e.g., 17E6 (Castel et al., Eur. J. Cell. Biol. 79(7):502-512, 2000); Mitjans et al., Int. J. Cancer 87(5):716-723, 2000)). In some embodiments, the monoclonal antibody is vedolizumab (e.g., Entyvio) or a variant thereof (Feagan et al., N. Engl. J. Med 369:699-710, 2013; Sandborn et al., N. Engl. J. Med. 369:711-721, 2013; Sands et al., Gastroenterology 147:618-627, 2014; and Milch et al., Neuroimmunol. 264:123-126, 2013; Wyant et al., J. Crohns Colitis 10(12):1437-1444, 2016; and Feagan et al., Gastroenterology 142(5):S160-S161, 2012).

[0821] In some embodiments, the antibody can be a Fab fragment of a monoclonal chimeric mouse-human antibody (e.g., abciximab (ReoPro, c7E3), Kononczuk et al., Curr. Drug Targets 16(13):1429-1437, 2015; Jiang et al., Appl. Microbiol. Biotechnol. 98(1):105-114, 2014), or a variant thereof. In some embodiments, the integrin antibody is a humanized monoclonal antibody. In some embodiments, the humanized monoclonal antibody is natalizumab (Tysabri) (Targan et al., Gastroenterology 132(5):1672-1683, 2007; Sandborn et al., N. Engl. J. Med. 353(18):1912-1925, 2005; Nakamura et al., Intern Med. 56(2):211-214, 2017; Singh et al., J. Pediatr. Gastroenterol. Nutr. 62(6):863-866, 2016). In some embodiments, the humanized monoclonal antibody is vitaxin (MEDI-523) or a variant thereof (Huveneers et al., Int, J. Radiat. Biol. 81(11-12):743-751, 2007; Coleman et al., Circ. Res. 84(11):1268-1276, 1999). In some embodiments, the humanized monoclonal antibody is etaracizumab (Abegrin, MEDI-522, LM609) or a variant thereof (Hersey et al., Cancer 116(6):1526-1534, 2010; Delbaldo et al., Invest New Drugs 26(1):35-43, 2008). In some embodiments, the humanized monoclonal antibody is CNTO95 (Intetumumab) or a variant thereof (Jia et al., Anticancer Drugs 24(3):237-250, 2013; Heidenreich et al., Ann. Oncol. 24(2):329-336, 2013; Wu et al., J. Neurooncol. 110(1):27-36, 2012). In some embodiments, the humanized monoclonal antibody is efalizumab (Raptiva) or a variant thereof (Krueger et al., J. Invest. Dermatol. 128(11):2615-2624, 2008; Li et al., PNAS 106(11):4349-4354, 2009; Woolacott et al., Health Technol. Assess 10:1-233, 2006). In some embodiments, the humanized monoclonal antibody is STX-100 (Stromedix) or a variant thereof (van Aarsen et al., Cancer Res. 68:561-570, 2008; Lo et al., Am. J. Transplant. 13(12):3085-3093, 2013). In some embodiments, the humanized monoclonal antibody is 264RAD or a variant thereof (Eberlein et al., Oncogene 32(37):4406-4417, 2013).

[0822] In some embodiments, the humanized monoclonal antibody is rovelizumab or a variant thereof (Goodman et al., Trends Pharmacol. Sci 33:405-412, 2012). In some embodiments, the humanized monoclonal antibody is Cytolin or a variant thereof (Rychert et al., Virology J. 10:120, 2013). In some embodiments, the humanized monoclonal antibody is etrolizumab or a variant thereof (Vermeire et al., Lancet 384:309-318, 2014; Rutgeerts et al., Gut 62:1122-1130, 2013; Lin et al., Gastroenterology 146:307-309, 2014; Ludviksson et al., J. Immunol. 162(8):4975-4982, 1999; Stefanich et al., Br. J. Pharmacol. 162(8):1855-1870, 2011). In some embodiments, the humanized monoclonal antibody is abrilumab (AMG 181; MEDI-7183) or a variant thereof (Pan et al., Br. J. Pharmacol. 169(1):51-68, 2013; Pan et al., Br. J. Clin. Pharmacol. 78(6):1315-1333, 2014). In some embodiments, the humanized monoclonal antibody is PF-00547659 (SHP647) or a variant thereof (Vermeire et al., Gut 60(8):1068-1075, 2011; Sandborn et al., Gastroenterology 1448(4):S-162, 2015). In some embodiments, the humanized monoclonal antibody is SAN-300 (hAQC2) or a variant thereof (Karpusas et al., J. Mol. Biol. 327:1031-1041, 2003). In some embodiments, the humanized monoclonal antibody is DI176E6 (EMD 5257) or a variant thereof (Goodman et al., Trends Pharmacol. Sci 33:405-412, 2012; and Sheridan et al., Nat. Biotech. 32:205-207, 2014).

[0823] In some embodiments, the integrin antibody is a chimeric monoclonal antibody. In some embodiments, the chimeric monoclonal antibody is volociximab or a variant thereof (Kuwada et al., Curr. Opin. Mol. Ther. 9(1):92-98, 2007; Ricart et al., Clin. Cancer Res. 14(23):7924-7929, 2008; Ramakrishnan et al., J. Exp. Ther. Oncol. 5(4):273-86, 2006; Bell-McGuinn et al., Gynecol. Oncol. 121:273-279, 2011; Almokadem et al., Exp. Opin. Biol. Ther. 12:251-7, 2012).

[0824] In some embodiments, the antibody specifically binds one or more (e.g., 1, 2, 3, 4, or 5) integrin. In some embodiments, the antibody specifically binds an integrin dimer (e.g., MLN-00002, MLNO2 (Feagan et al., Clin. Gastroenterol. Hepatol. 6(12):1370-1377, 2008; Feagan et al., N. Engl. J. Med. 352(24):2499-2507, 2005). In certain embodiments, the antibody comprises or consists of an antigen-binding fragment of abciximab (Reopro) (Straub et al., Eur. J. Cardiothorac Surg. 27(4):617-621, 2005; Kim et al., Korean J. Intern. Med. 19(4):220-229, 2004). In some embodiments, the integrin inhibitor is an antibody-drug conjugate (e.g., IMGN388 (Bendell et al., EJC Suppl 8(7):152, 2010).

[0825] Further examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 5,919,792; 6,214,834; 7,074,408; 6,833,373; 7,655,624; 7,465,449; 9,558,899; 7,659,374; 8,562,986; 8,398,975; and 8,853,149; US 2007/0117849; US 2009/0180951; US 2014/0349944; US 2004/0018192; WO 11/137418; and WO 01/068586; each of which is incorporated by reference in its entirety.

[0826] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0827] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9M, about 0.510.sup.9 M, about 110.sup.10M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0828] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0829] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1(inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1(inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1(inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins

[0830] In some embodiments, the integrin inhibitor is a fusion protein (e.g., an Fc fusion protein of an extracellular domain of an integrin or an integrin receptor), a soluble receptor (e.g., the extracellular domain of an integrin or an integrin receptor), or a recombinant integrin binding protein (e.g., an integrin ligand). See, e.g., Lode et al., PNAS 96(4):1591-1596, 1999; Stephens et al., Cell Adhesion Comm. 7:377-390, 2000; and US 2008/0739003; incorporated by reference herein). Non-limiting examples of fusion proteins that are integrin inhibitors include Ag25426 (Proteintech).

Small Molecules Antagonists

[0831] In some embodiments, the integrin inhibitor is a small molecule. In some embodiments, the small molecule is a non-peptide small molecule. In some embodiments, the non-peptide small molecule is a RGD (ArgGlyAsp)-mimetic antagonist (e.g., tirofiban (Aggrastat); Pierro et al., Eur. J. Ophthalmol. 26(4):e74-76, 2016; Guan et al., Eur. J. Pharmacol 761:144-152, 2015. In some embodiments, the small molecule is 4 antagonist (e.g., firategrast (Miller et al., Lancet Neurol. 11(2):131-139, 2012) AJM300 (Yoshimura et al., Gastroenterology 149(7):1775-1783, 2015; Takazoe et al., Gastroenterology 136(5):A-181, 2009; Sugiura et al., J. Crohns Colitis 7(11):e533-542, 2013)). In some embodiments, the small molecule is 41 antagonist (e.g., IVL745 (Norris et al., J. Allergy Clin. Immunol. 116(4):761-767, 2005; Cox et al., Nat. Rev. Drug Discov. 9(10):804-820, 2010)), BIO-1211 (Abraham et al., Am. J. Respir. Crit. Care Med 162:603-611, 2000; Ramroodi et al., Immunol. Invest. 44(7):694-712, 2015; Lin et al., J. Med Chem. 42(5):920-934, 1999), HMR 1031 (Diamant et al., Clin. Exp. Allergy 35(8):1080-1087, 2005); valategrast (R411) (Cox et al., Nat. Rev. Drug Discov. 9(10):804-820, 2010), GW559090X (Ravensberg et al., Allergy 61(9):1097-1103, 2006), TR14035 (Sircar et al., Bioorg. Med Chem. 10(6):2051-2066, 2002; Cortijo et al., Br. J. Pharmacol. 147(6):661-670, 2006)). In some embodiments, the small molecule is v antagonist (e.g., L0000845704, SB273005). In some embodiments, the small molecule is 51 antagonist (e.g., JSM6427). In some embodiments, the small molecule is GLPG0974 (Vermeire et al., J. Crohns Colitis Suppl. 1:S39, 2015). In some embodiments, the small molecule is MK-0429 (Pickarksi et al., Oncol. Rep. 33(6):2737-45, 2015; Rosenthal et al., Asia Pac J. Clin. Oncol. 6:42-8, 2010). In some embodiments, the small molecule is JSM-6427 or a variant thereof (Zahn et al., Arch. Ophthalmol. 127(10):1329-1335, 2009; Stragies et al., J. Med Chem. 50:3786-94, 2007).

[0832] In some embodiments, the small molecule integrin inhibitor can be PTG-100, which is described in, e.g., Shames et al., Pharmakokinetics and Pharmacodynamics of the Novel Oral Peptide Therapeutic PTG-100(47 Integrin Antagonist) in Normal Healthy Volunteers, 24.sup.th United European Gastroentrology Week, October 15-19, Vienna, Austria, 2016.

[0833] In some embodiments, the small molecule targets 2 integrin. In some embodiments, the small molecule is SAR-118(SAR1118) or a variant thereof (Zhong et al., ACS Med. Chem. Lett. 3(3):203-206, 2012; Suchard et al., J. Immunol. 184:3917-3926, 2010; Yandrapu et al., J. Ocul. Pharmacol. Ther: 29(2):236-248, 2013; Semba et al., Am. J. Ophthalmol. 153:1050-60, 2012). In some embodiments, the small molecule is BMS-587101 or a variant thereof (Suchard et al., J. Immunol. 184(7):3917-3926, 2010; Potin et al., J. Med Chem. 49:6946-6949, 2006). See e.g., Shimaoka et al., Immunity 19(3):391-402, 2003; U.S. Pat. Nos. 7,138,417; 7,928,113; 7,943,660; and 9,216,174; US2008/0242710; and US 2008/0300237.

[0834] In some embodiments, the integrin inhibitor is an inhibitor as shown in the following table:

TABLE-US-00019 Target- Drug based Name Actions Structure Other Drug Names References ALPHA-4 INHIBITORS ELND-004 Elan Corp plc CD49d antagonist Alpha 4 inhibitor [00001] ELND-004 Soler-Ferran, Dulce, and Michael J Briskin. Integrin 47 Antagonists: Activities, Mechanisms of Action and Therapeutic Prospects. Current Immunology Reviews 8.2 (2012): 118-134. and U.S. Pat. No. 6,436,904; U.S. Pat. No. 6,492,421; U.S. Pat. No. 6,903,088; U.S. Pat. No. 6,939,855; U.S. Pat. No. 6,949,570; U.S. Pat. No. 7,030,114; U.S. Pat. No. 7,115,768; U.S. Pat. No. 7,166,580; U.S. Pat. No. 7,320,960; U.S. Pat. No. 7,452,912; U.S. Pat. No. 7,335,663 ELND-002 CD49d ELND-002; U.S. Pat. No. 6,436,904; (PEGylated antagonist ELND-002 U.S. Pat. No. 6,492,421; subcutaneous (PEGylated U.S. Pat. No. 6,903,088; formulation, subcutaneous U.S. Pat. No. 6,939,855; hematological formulation, U.S. Pat. No. 6,949,570; malignancies/ hematological U.S. Pat. No. 7,030,114; multiple malignancies/multiple U.S. Pat. No. 7,115,768; sclerosis) sclerosis), Elan; alpha- U.S. Pat. No. 7,166,580; Elan Corp 4 integrin inhibitor U.S. Pat. No. 7,320,960; plc (injectable, U.S. Pat. No. 7,452,912; hematologic U.S. Pat. No. 7,335,663 malignancies), Elan ELND-002 (oral, multiple sclerosis) Elan Corp plc CD49d antagonist [00002] ELND-002; ELND-002 (oral, multiple sclerosis), Elan; alpha- 4 integrin antagonists (oral, autoimmune diseases), Elan; alpha-4 integrin antagonists (oral, multiple sclerosis), U.S. Pat. No. 6,436,904; U.S. Pat. No. 6,492,421; U.S. Pat. No. 6,903,088; U.S. Pat. No. 6,939,855; U.S. Pat. No. 6,949,570; U.S. Pat. No. 7,030,114; U.S. Pat. No. 7,115,768; U.S. Pat. No. 7,166,580; U.S. Pat. No. 7,320,960; U.S. Pat. No. 7,452,912; Elan U.S. Pat. No. 7,335,663 alpha4- beta1/alpha4- beta7 antagonists (asthma), Roche Integrin alpha- 4/beta-1 antagonist; Integrin alpha- 4/beta-7 antagonist [00003] alpha4-beta1/alpha4- beta7 antagonist prodrugs (asthma), Roche; alpha4- beta1/alpha4-beta7 antagonists (asthma), Roche See chemical structure; Sidduri A et al. Identification of N- acyl 4-(5-pyrimidine-2,4- dionyl)phenylalanine derivatives and their orally active prodrug esters as dual-acting alpha4-beta1 and alpha4-beta7 receptor antagonists Bioorganic and Medicinal Chemistry Letters (2013) 23 (4) 1026-1031 alpha- 4/beta-7 integrin modulators (IBD), Morphic Therapeutic Integrin alpha- 4/beta-7 modulator [00004] alpha-4/beta-7 integrin modulators (IBD), Morphic Therapeutic See chemical structure ET-3764 Integrin ET-3764; integrin PCT/CA2017/000244, integrin alpha- alpha-4-beta-7- which published as alpha-4- 4/beta-7 targeting nacellin (oral, WO2018085921A1 beta-7- antagonist IBD), Encycle; targeting macrocyclic nacellin peptidomimetics, (oral, IBD), Encycle; nacellins Encycle (peptidomimetic Therapeutics macrocycles), Inc Encycle E-6007 Integrin E-6007; integrin See chemical structure; (ER- antagonist activation inhibitor Ohkuro M, et al. E6007: An 46419501) (ulcerative orally active inhibitor of Eisai Co colitis/Crohn's integrin activation for Ltd disease), EA Pharma; inflammatory bowel integrin activation disease inhibitor (ulcerative 2007 (May 20) Abs-S1588; colitis/Crohn's Digestive Disease Week disease), Eisai ER- 464195-01 integrin (analogue of E6007) EA Pharma Co., Ltd inhibits integrin activation by dissociating interaction between calreticulin (CRT) and integrin 4 (ITGA4) [00005] Ohkuro, Masayoshi, et al. Calreticulin and integrin alpha dissociation induces anti-inflammatory programming in animal models of inflammatory bowel disease. Nature communications 9.1 (2018): 1982 and WO2005063705 HCA-3551 Integrin HCA-3551; Hirano, Yuta, et al. EA Pharma alpha- alpha-4 Ameliorating effects of HCA Co Ltd 4/beta-1 integrin antagonist 3551, alpha 4 integrin antagonist; (multiple sclerosis), antagonist, on Theiler's Integrin Ajinomoto murine encephalomyelitis alpha- virus (TMEV)-induced 4/beta-7 demyelinating disease. antagonist Journal of Neuroimmunology 275.1 (2014): 157. DW-908e Pharmacopeia Inc Integrin alpha- 4/beta-1 antagonist [00006] DW-908e; PS-181895; PS-460644; PS- 489655; PS-969819; allergy therapeutics, Daiichi; asthma therapeutics, Daiichi; asthma/allergy therapeutics, Daiichi See chemical structure VLA-4 antagonists, Texas/ Schering- Plough Encysive Pharma- ceuticals Inc Integrin alpha- 4/beta-1 antagonist [00007] AVA-4746; TBC-3342; TBC-4746; VLA-4 antagonists, Encysive/Schering- Plough; VLA-4 antagonists, Texas/Schering- Plough; integrin alpha- 4/beta-1 antagonists, Schering- Plough/Encysive See chemical structure and WO-2004044046; EP-01203766; WO-2010008719 GW- 559090 GlaxoSmith Kline plc Integrin alpha- 4/beta-1 antagonist [00008] 559090; GW-559090; alpha-4 integrin antagonist (inhaled), GlaxoSmithKline See chemical structure and WO-00037444 TRK-170 Integrin TRK-170 Koga Y, Kainoh, M Effect of Toray alpha- an orally active small Industries 4/beta-1 molecule Inc antagonist; alpha4beta1/alpha4beta7 Integrin integrin antagonist, TRK- alpha- 170, on experimental colitis 4/beta-7 in mice International antagonist Congess on Immunology; 2010 (August 22-27) and Hiroe Hirokawa, et al. Inhibitory Effects of an Orally Active Small Molecule Alpha4beta1/Alpha4beta7 Integrin Antagonist, Trk-170, on Spontaneous Colitis in HLA-B27 Transgenic Rats Digestive Disease Week; 2014 (May 05) Abs Mo1706 integrin Integrin carotegrast; integrin WO-00216329 antagonists alpha- antagonists (inflammation), 4/beta-1 (inflammation), Ajinomoto antagonist Ajinomoto Co Inc TRK-720 Toray Industries Inc Integrin alpha- 4/beta-1 antagonist [00009] TRK-720; TRK-720 hydrate; VLA4 inhibitor (inhaled, asthma), Toray Shiraki M Physical characterization of trk-720 hydrate, the very late antigen-4 (via-4) inhibitor, as a solid form for inhalation: preparation of the hydrate by solvent exchange among its solvates and mechanistical considerations Journal of Pharmaceutical Sciences; (2010) 99 (9) 3986-4004 VLA-4 Integrin MK-0617 and [MK-0617] CARLEVARO, C. antagonists alpha- MK-0668 MANUEL, et al. Plausible (inflammatory 4/beta-1 binding mode of the active disorders) antagonist 41 antagonist, Mk-0617, Merck determined by docking and free energy calculations. Journal of Theoretical and Computational Chemistry 12.02 (2013): 1250108. and [MK-0668] Lin, Linus S., et al. Discovery of N-{N-[(3- Cyanophenyl) sulfonyl]-4 (R)-cyclobutylamino-(I)- prolyl}-4-[(3,5- dichloroisonicotinoyl) amino]-(I)-phenylalanine (MK-0668), an Extremely Potent and Orally Active Antagonist of Very Late Antigen-4. Journal of medicinal chemistry 52.11 (2009): 3449-3452. LFA- CD11a LFA-1/ICAM WO-2005105766 1/ICAM modulator; interaction inhibitors, interaction ICAM-1 Biogen Idec inhibitors modulator Biogen Inc alpha-4 Integrin alpha-4 beta-1/alpha-4 Lawson et al. Selection of a beta- alpha- beta-7 integrin 2-azabicyclo[2.2.2]octane- 1/alpha-4 4/beta-1 antagonists, Johnson based .alpha.(4).beta.(1) beta-7 antagonist; & Johnson integrin antagonist as an integrin Integrin inhaled anti-asthmatic agent antagonists alpha- Bioorganic and Medicinal Johnson & 4/beta-7 Chemistry; 2006 14 (12) Johnson antagonist 4208-4216 VLA- 4/VCAM antagonists (inflammation) Elan Corp plc Integrin alpha- 4/beta-1 antagonist; Vascular cell adhesion protein 1 modulator [00010] CT-737; CT-747; CT- 757; CT-767; VLA- 4/VCAM antagonists (inflammation), Elan/Wyeth; VLA- 4/VCAM antagonists (inflammation), Elan/Wyeth-Ayerst Xu Y, et al Arylsulfonamide pyrimidines as vla-4 antagonists Bioorganic and Medicinal Chemistry Letters; 2013 23 (10) 3070- 3074 integrin antagonists, Zeneca Group plc Integrin antagonist; Vascular cell adhesion protein 1 antagonist [00011] VCAM antagonists, Zeneca; fibronectin antagonists, Zeneca; integrin antagonists, Zeneca WO-09702289 and Haworth D et al., Anti- inflammatory activity of c(ILDV-NH(CH2)5CO), a novel, selective, cyclic peptide inhibitor of VLA-4- mediated cell adhesion British Journal of Pharmacology; 1999 126 (8) 1751-1760 SB-683698 Integrin SB-683698; Tsuda-Tsukimoto M, et al. Tanabe alpha- TR-14035; Characterization of Seiyaku Co 4/beta-1 TR-9109 hepatobiliary transport Ltd antagonist; systems of a novel Integrin alpha4beta1/alpha4beta7 alpha- dual antagonist, TR-14035; 4/beta-7 Pharmaceutical Research; antagonist 2006 23 (11) 2646-2656 R-1541 Roche Holding Co Integrin antagonist [00012] R-1541 See chemical structure alpha- 4/beta-7 antagonists, Millennium LeukoSite Inc Integrin alpha- 4/beta-7 antagonist; MAdCAM inhibitor [00013] A4B7 program, Millennium; alpha- 4/beta-7 antagonists, Millennium; autoimmune therapeutics, Genzyme/LeukoSite; beta-7 integrin receptor antagonists, LeukoSite; Harriman GC et al. Cell adhesion antagonists: Synthesis and evaluation of a novel series of phenylalanine based inhibitors Bioorganic and Medicinal Chemistry Letters; 2000 10 (14) 1497- 1499 inflammatory disease therapeutics, Genzyme/LeukoSite; inflammatory therapeutics, LeukoSite; small molecule IBS program, Millennium CP-664511 Integrin CP-664511; VLA-4 WO-00151487 Pfizer alpha- antagonists, Pfizer and 4/beta-1 Kudlacz E, et al. antagonist; Pulmonary eosinophilia in a Vascular murine model of allergic cell inflammation is attenuated adhesion by small molecule protein 1 alpha4beta1 antagonists antagonist Journal of Pharmacology and Experimental Therapeutics; 2002 301 (2) 747-752 BETA-7 INHIBITORS alpha Integrin alpha epsilon beta 7 US 09/856,544 and epsilon alpha-E integrin antagonists PCT/US99/27817 beta 7 antagonist; (inflammatory integrin Integrin disease), NIAID antagonists beta-7 (inflammatory antagonist disease), NIAID alpha- Integrin alpha-4/beta-7 integrin Callier Dublanchet A-C, 4/beta-7 alpha- inhibitors, Jouveinal et al. Potential alpha4beta7 integrin 4/beta-7 integrin-mediated cell inhibitors, antagonist adhesion inhibitors: Institut de synthesis and evaluation of Recherche novel pyrazolones Jouveinal derivatives and a study of SA (IRJ) their stability ACS Meeting; 2000 220th (Washington DC) MEDI 142 ITGAL-Integrin alpha-L lifitegrast Sunesis Pharma- ceuticals Inc CD11a antagonist (integrin alpha-L (ITGAL)) ICAM-1 inhibitor [00014] SAR-1118; SHP-606; SPD-606; Xiidra; dual LFA-1/ICAM-1 inhibitors (inflammatory diseases), SARcode; dual LFA-1/ICAM-1 inhibitors, Sunesis; Zhong M, et al. Structure- activity relationship (SAR) of the alpha-amino acid residue of potent tetrahydroisoquinoline (THIQ)-derived LFA- 1/ICAM-1 antagonists Bioorganic and Medicinal lifitegrast; lifitegrast Chemistry Letters; 2011 21 sodium (1) 307-310 BIRT-2584 CD11a BIRT-0377; BIRT- Wu J-P, et al. The Boehringer antagonist; 2584; BIRT-2584 XX; discovery of 1H- Ingelheim Cell BIRT-377; ICAM-1 imidazo[1,2-alfa]imidazol-2- International adhesion antagonists one derivatives as LFA-1 GmbH molecule (inflammation), inhibitors Inflammation inhibitor; Boehringer Ingelheim; Research; 2003 52 (Suppl ICAM-1 LFA-1 inhibitors 2) Abs 137 inhibitor; (inflammation/allergy), and ITGAL Boehringer Ingelheim WO-2007084882 LFA- CD11a CP151088; ICAM- Khojasteh SC, et al. 1/ICAM-1 modulator; 2078; ICAM-850; LFA- Preclinical absorption, interaction CD11b 1/ICAM-1 interaction distribution, metabolism and inhibitors, modulator; inhibitors, excretion (ADME) Genentech ICAM-1 Genentech/Roche; characterization of Inc modulator LFA-1/ICAM-1 ICAM1988, an LFA-1/ICAM interaction inhibitors, antagonist, and its prodrug Roche; Mac-1/ICAM-1 Xenobiotica; 2008 38 (3) interaction inhibitors, 340-352 Genentech; Mac- and 1/ICAM-1 interaction WO-02059114 inhibitors, Roche; RO-0276845; RO-5182851; RO-5184438; RO-5200045 LFA-1 inhibitors, Novartis Pharma AG CD11a antagonist; ICAM inhibitor [00015] LFA-1 inhibitors, Novartis; LFA-451; LFA-703; LFA-878; XVA-143 See chemical structure IC-776 ICOS Corp CD11a antagonist; ICAM inhibitor [00016] A-276594; A-286982; A-292949; A-295339; A-324920; IC-52593; IC-776; LFA-1 antagonists, ICOS; LFA-1/ICAM-1 inhibitors, Abbott/ICOS; LFA- 1/ICAM-1 inhibitors, ICOS; LFA-1/ICAM-1 inhibitors, ICOS/Biogen Pei Z, et al. Discovery of potent antagonists of leukocyte function- associated antigen- 1/inercellular adhesion molecule-1 interaction. 3. Amide (C-ring) structure- activity relationship and improvement of overall properties of arylthio cinnamides Journal of Medicinal Chemistry; 2001 44 (18) 2913-2920 [00017]
Other exemplary integrin inhibitors include the following: [0835] SMART anti-L-selectin Mab from PDL BioPharma Inc., which is L-Selectin antagonist, and described in WO-09706822, and Co M S, et al. Properties and pharmacokinetics of two humanized antibodies specific for L-selectin; Immunotechnology; 1999 4253-266; both of which are hereby incorporated by reference [0836] SEL-K2, an anti-PSGL-1 antibody, from Tetherex Pharmaceuticals Ic, which is described in Barbara Muz, et al. Inhibition of P-Selectin and PSGL-1 Using Humanized Monoclonal Antibodies Increases the Sensitivity of Multiple Myeloma Cells to Proteasome Inhibitors American Society of Hematology Annual Meeting and Exposition; 2014 56th (December 08) Abs 4758, which is hereby incorporated by reference [0837] Vatelizumab described in I. A. Antonijevic, et al. Safety, tolerability and pharmacodynamic characterization of vatelizumab, a monoclonal antibody targeting very-late-antigen (VLA)-2: a randomized, double-blind, placebo-controlled phase 1 study Abstract release date: Sep. 23, 2015) ECTRIMS Online Library. Oct. 9, 2015; and WO-2010095031; WO-2011104604; WO-2010052556, which are all hereby incorporated by reference [0838] anti-VCAM mAb, which is described in Soriano, Antonio, et al. VCAM-1, but not ICAM-1 or MAdCAM-1, immunoblockade ameliorates DSS-induced colitis in mice. Laboratory investigation 80.10 (2000): 1541; and Gerritsen M E, et al. (1995). Activation-dependent isolation and culture of murine pulmonary microvascular endothelium. Microcirculation 2:151-163.

Cyclic Peptides

[0839] In some embodiments, the integrin inhibitor is a cyclic peptide. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence as set forth in the amino acid sequence of a ligand recognition sequence of an endogenous integrin ligand. In some embodiments, the cyclic peptide competes for a target integrin ligand binding site with an endogenous integrin ligand. In some embodiments, the cyclic peptide includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8) D-amino acids. In some embodiments, the cyclic peptide is a synthetic cyclic peptide. In some embodiments, the synthetic cyclic peptide is a heptapeptide. In some embodiments, the synthetic cyclic peptide is eptifabitide (Integrilin), or a variant thereof. In some embodiments, the cyclic peptide comprises a heterocyclic nucleic (e.g., a benzodiazepinone, a piperazine, a benzoazepinone, a nitroaryl, an isoxazoline, an indazole, or a phenol; Spalluto et al., Curr. Med. Chem. 12:51-70, 2005). In some embodiments, the cyclic peptide is a macrocycle (see, e.g., Halland et al., ACS Med. Chem. Lett. 5(2):193-198, 2014). In some embodiments, the peptide is ALG-1001 or a variant thereof (Mathis et al., Retin. Phys. 9:70, 2012). In some embodiments, the cyclic peptide is an imidazolone-phenylalanine derivative, a heteroaryl, hetrocyclic, and aryl derivative, a bicyclic-aromatic amino acid derivative, a cyclohexane-carboxylic acid derivative, a di-aryl substituted urea derivative, a multimeric L-alanine derivative, a L-alanine derivative, or a pyrimidyl-sulfonamide derivative (see, e.g., U.S. Pat. Nos. 6,630,492; 6,794,506; 7,049,306; 7,371,854; 7,759,387; 8,030,328; 8,129,366; 7,820,687; 8,350,010; and 9,345,793).

Peptidomimetics

[0840] In some embodiments, the integrin inhibitor is a peptidomimetic. In some embodiments, the peptidomimetic has an integrin-ligand recognition motif (e.g., RGD, KTS, or MLD). See, e.g., Carron et al., Cancer Research 58:1930-1935, 1998; Fanelli et al., Vascular Cell 6:11, 2014; and De Marco et al., Curr. Top. Med. Chem. 16(3):343-359, 2016.

[0841] In some embodiments, the peptidomimetic is an RGD(ArgGlyAsp)-based peptide (U.S. Pat. No. 8,809,338, incorporated by reference in its entirety herein). In some embodiments, the RGD-based peptide can be cilengitide or a variant thereof (EMD 12974) (Mas-Moruno et al., Anticancer Agents Med. Chem. 10:753-768, 2010; Reardon et al., Future Oncol. 7(3):339-354, 2011; Beekman et al., Clin. Genitourin Cancer 4(4):299-302, 2006; SC56631 (e.g., Engleman et al., Am Soc. Cln. Invest. 99(9):2284-2292, 1997; Peng et al., Nature Chem Biol. 2:381-389, 2006). In some embodiments, the peptidomimetic can be a Lys-Gly-Asp (KGD)-based peptide. In some embodiments, the peptidomimetic can be vipegitide or a variant thereof (Momic et al., Drug Design Devel. Therapy 9:291-304, 2015). In some embodiments, the peptidomimetic can be a peptide conjugated with an antimicrobial synthetic peptide. (e.g., ACDCRGDCFC conjugated with (KLAKLAK).sub.2(Ellerby et al., Nat. Med. 5(9):1032-1038, 1999). See, e.g., U.S. Pat. No. 8,636,977.

Disintegrins

[0842] In some embodiments, the integrin inhibitor can be a disintegrin. The term disintegrin as used herein refers to a low molecular weight peptide integrin inhibitor derived from a snake venom (e.g., pit viper venom). In some embodiments, the disintegrin is a RGD(ArgGlyAsp)-, a KTS- or an MLD-based disintegrin.

[0843] Non-limiting examples of disintegrins include accutin, accurhagin-C, albolabrin, alternagin-c, barbourin, basilicin, bitisgabonin-1, bitisgabonin-2, bitistatin, cerastin, cereberin, cumanastatin 1, contortrostatin, cotiarin, crotatroxin, dendroaspin, disba-01, durissin, echistatin, EC3, elegantin, eristicophin, eristostatin, EMS11, EO4, EO5, flavoridin, flavostatin, insularin, jarastatin, jerdonin, jerdostatin, lachesin, lebein (e.g., lebein-1, lebein-2), leberagin-C, lebestatin, lutosin, molossin, obtustatin, ocellatusin, rhodocetin, rhodostomin, R-mojastin 1, salmosin, saxatilin, schistatin, tablysin-15, tergeminin, triflavin, trigramin, trimestatin, VA6, vicrostatin, viridin, viperstatin, VB7, VLO4, and VLO5, or a variant thereof. See, e.g., Arruda Macedo et al., Curr. Protein. Pept. Sci. 16(6):532-548, 2015; Hsu et al., Sci. Rep. 6:23387, 2016; Kele et al. Curr. Protein Pept. Sci. 6:532-548, 2015; Koh et al., Toxicon 59(4):497-506, 2012; Scarborough et al., J. Biol. Chem. 268:1058-1065, 1993; Kisiel et al., FEBS Lett. 577:478-482, 2004; Souza et al., Arch. Biochem. Biophys. 384:341-350, 2000; Eble et al., J. Biol. Chem. 278:26488-26496, 2003; Marcinkiewicz et al., J. Biol. Chem. 274:12468-12473, 1999; Calvete et al., J. Proteome Res. 6:326-336, 2007; Scibelli et al., FEMS Microbiol. Lett. 247:51-57, 2005; Oliva et al., Toxicon 50:1053-1063, 2007; Minea et al., Toxicon 59:472-486, 2012; Smith et al., FEBS Lett. 512:111-115, 2002; Tselepis et al., J. Biol. Chem. 272:21341-21348, 1997; Da Silva et al., Tromb. Res. 123:731-739, 2009; Thibault et al., Mol. Pharmacol. 58:1137-1145, 2000; Lu et al., Biochem. J 304:818-825, 1994; Yeh et al., Biochim. Biophys. Acta. 1425:493-504, 1998; Huang et al., Exp. Hematol. 36:1704-1713, 2008; Shih et al., Matrix Biol. 32:152-159, 2013; Wang et al., Br. J. Pharmacol. 160:1338-1351, 2010; Della-Casa et al., Toxicon 57:125-133, 2011; Sheu et al., Biochim. Biophys. Acta. 1336:445-454, 1997; Fujii et al., J. Mol. Biol. 332:115-122, 2003; Bilgrami et al., J. Mol. Biol. 341:829-837, 2004; Zhou et al., Toxicon 43:69-75, 2004; Scarborough et al., J. Biol. Chem. 268:1066-1073, 1993; Shebuski et al., J. Biol. Chem. 264:21550-21556, 1989; Lu et al., Biochem. J 304:929-936, 1994; McLane et al., Biochem. J. 301:429-436, 1994; Juarez et al., Toxicon 56:1052-1058, 2010; Olfa et al., Lab. Invest. 85:1507-1516, 2005; Elbe et al., Matrix Biol. 21:547-558, 2002; Bazan-Socha et al., Biochemistry 43:1639-1647, 2004; Danen et al., Exp. Cell. Res. 238:188-196, 1998; Marcinkiewicz et al., Biochemistry 38(40):13302-13309, 1999; Calvete et al., Biochem. J. 372:725-734, 2003; Swenson et al., Pathophysiol. Haemost. Thromb. 34:169-176, 2005; Kwon et al., PLoS One 8; e81165, 2013; Yang et al., Toxicon 45:661-669, 2005; Limam et al., Matrix Biol. 29:117-126, 2010; Gan et al., J. Biol. Chem. 263:19827-19832, 1988; Ma et al., Thromb. Haemost. 105(6):1032-1045, 2011; and U.S. Pat. No. 7,074,408, incorporated in their entirety herein.

Chemokine/Chemokine Receptor Inhibitors

[0844] The term chemokine/chemokine receptor inhibitors refers to an agent which decreases the ability of a chemokine to bind to its receptor, where the chemokine is one of CXCL10 (IL-10), CCL11, or an ELR chemokine, or the chemokine receptor is CCR2 or CCR9.

CXCL10 (IP-10) Inhibitors

[0845] As used herein CXCL10, interferon gamma-induced protein 10 and IP-10 can be used interchangeably. CXCL10 binds to the CXCR3 receptor (e.g., CXCR3-A or CXCR3-B).

[0846] The term CXCL10 inhibitor refers to an agent which decreases the ability of CXCL10 to bind to a CXCR3 receptor (e.g., CXCR3-A and/or CXCR3-B).

[0847] In some embodiments, the CXCL10 inhibitor can decrease the binding between CXCL10 and CXCR3-A by blocking the ability of CXCL10 to interact with CXCR3-A. In some embodiments, the CXCL10 inhibitor can decrease the binding between CXCL10 and CXCR3-B by blocking the ability of CXCL10 to interact with CXCR3-B.

[0848] In some instances, the CXCL10 inhibitor that decreases the binding between CXCL10 and a CXCR3 (e.g., CXCR3-A and/or CXCR3-B) is a small molecule. In some instances, the CXCL10 inhibitor that decreases the binding between CXCL10 and a CXCR3 (e.g., CXCR3-A and/or CXCR3-B) is an antibody or an antigen-binding antibody fragment. In some instances, the CXCL10 inhibitor that decreases the binding between CXCL10 and a CXCR3 (e.g., CXCR3-A and/or CXCR3-B) is a peptide (e.g., a peptide antagonist of a CXCR3 receptor, e.g., one or both of CXCR-A and/or CXCR-B).

[0849] Exemplary sequences for human CXCL10 and human CXCR3 are shown below.

Human CXCL10 (SEQ ID NO: 182)

[0850] vplsrtvrc tcisisnqpv nprsleklei ipasqfcprv eiiatmkkkg ekrclnpesk aiknllkavs kerskrsp

Human CXCR3 Isoform 1 (SEQ ID NO: 183)

[0851] mvlevsdhqv lndaevaall enfsssydyg enesdsccts ppcpqdfsln fdraflpaly
sllfllgllg ngavaavlls rrtalsstdt fllhlavadt llvltlplwa vdaavqwvfg
sglckvagal fninfyagal llacisfdry lnivhatqly rrgpparvtl tclavwglcl
lfalpdfifl sahhderlna thcqynfpqv grtalrvlql vagfllpllv maycyahila
vllvsrgqrr lramrlvvvv vvafalcwtp yhlvvlvdil mdlgalarnc gresrvdvak
svtsglgymh cclnpllyaf vgvkfrermw mlllrlgcpn qrglqrqpss srrdsswset
seasysgl

Human CXCR3 Isoform 2 (SEQ ID NO: 184)

[0852] melrkygpgr lagtviggaa qsksqtksds itkeflpgly tapsspfpps qvsdhqvlnd
aevaallenf sssydygene sdscctsppc pqdfslnfdr aflpalysll fllgllgnga
vaavllsrrt alsstdtfll hlavadtllv ltlplwavda avqwvfgsgl ckvagalfni
nfyagallla cisfdrylni vhatqlyrrg pparvtltcl avwglcllfa lpdfiflsah
hderlnathc qynfpqvgrt alrvlqlvag fllpllvmay cyahilavll vsrgqrrlra
mrlvvvvvva falcwtpyhl vvlvdilmdl galarnegre srvdvaksvt sglgymhccl
npllyafvgv kfrermwmll lrlgcpnqrg lqrqpsssrr dsswsetsea sysgl

CXCL10 InhibitorsAntibodies

[0853] In some embodiments, the CXCL10 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CXCL10 or a CXCR3 receptor (e.g., CXCR3-A and/or CXCR3-B), or both a CXCL10 and a CXCR3 receptor (e.g., CXCR3-A and/or CXCR3-B). In some embodiments, a CXCL10 inhibitor can bind to both CXCR3-A and CXCR3-B.

[0854] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; and Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0855] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0856] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; and Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0857] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; and Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety (e.g., the sections describing CXCL10 inhibitors).

[0858] In other instances, the CXCL10 inhibitor is a monoclonal antibody (mAb) (see, e.g., WO05/58815). For example, the CXCL10 inhibitor can be Eldelumab (MDX-1100 or BMS-936557), BMS-986184 (Bristol-Meyers Squibb), or NI-0801 (NovImmune). See, e.g., Kuhne et al., J. Immunol. 178(1):S241, 2007; Sandborn et al., J. Crohns Colitis 11(7):811-819, 2017; and Danese et al., Gastroenterology 147(5):981-989, 2014. Additional examples of CXCL10 inhibitors that are antibodies are described in U.S. Patent Application Publication Nos. 2017/0158757, 2017/0081413, 2016/0009808, 2015/0266951, 2015/0104866, 2014/0127229, 2014/0065164, 2013/0216549, 2010/0330094, 2010/0322941, 2010/0077497, 2010/0021463, 2009/0285835, 2009/0169561, 2008/0063646, 2005/0191293, 2005/0112119, 2003/0158392, 2003/0031645, and 2002/0018776; and WO 98/11218, each of which is incorporated by reference in its entirety (e.g., the description of CXCL10 inhibitors).

[0859] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0860] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0861] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0862] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1 s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0863] Additional examples of CXCL10 inhibitors that are antibodies or antigen-binding antibody fragments are known in the art.

CCL11 Inhibitors

[0864] The term CCL11 inhibitor refers to an agent which decreases the ability of CCL11 to bind to one or more of CCR2, CCR3, and CCR5.

[0865] In some embodiments, the CCL11 inhibitor can decrease the binding between CCL11 and CCR2 by blocking the ability of CCL11 to interact with CCR2. In some embodiments, the CCL11 inhibitor can decrease the binding between CCL11 and CCR3 by blocking the ability of CCL11 to interact with CCR3. In some embodiments, the CCL11 inhibitor can decrease the binding between CCL11 and CCR5 by blocking the ability of CCL11 to interact with CCR5.

[0866] In some embodiments, a CCL11 inhibitor is an antibody or an antigen-binding fragment thereof.

[0867] Exemplary sequences for human CCL11, human CCR2, human CCR3, and human CCR5 are shown below.

TABLE-US-00020 HumanCCL11 (SEQIDNO:185) mkvsaallwllliaaafspqglagpasvpttccfnlanrk iplqrlesyrritsgkcpqkavifktklakdicadpkkkw vqdsmkyldqksptpkp HumanCCR2IsoformA (SEQIDNO:186) mlstsrsrfirntnesgeevttffdydygapchkfdvkqi gaqllpplyslvfifgfvgnmlvvlilinckklkcltdiy llnlaisdllflitlplwahsaanewvfgnamcklftgly higyfggiffiilltidrylaivhavfalkartvtfgvvt svitwlvavfasvpgiiftkcqkedsvyvcgpyfprgwnn fhtimrnilglvlpllimvicysgilktllrcrnekkrhr avrviftimivyflfwtpynivillntfqeffglsncest sqldqatqvtetlgmthccinpiiyafvgekfrslfhial gcriaplqkpvcggpgvrpgknvkvttqglldgrgkgksi grapeaslqdkega HumanCCR2IsoformB (SEQIDNO:187) mlstsrsrfirntnesgeevttffdydygapchkfdvkqi gaqllpplyslvfifgfvgnmlvvlilinckklkcltdiy llnlaisdllflitlplwahsaanewvfgnamcklftgly higyfggiffiilltidrylaivhavfalkartvtfgvvt svitwlvavfasvpgiiftkcqkedsvyvcgpyfprgwnn fhtimrnilglvlpllimvicysgilktllrcrnekkrhr avrviftimivyflfwtpynivillntfqeffglsncest sqldqatqvtetlgmthccinpiiyafvgekfrrylsvff rkhitkrfckqcpvfyretvdgvtstntpstgeqevsagl HumanCCR3Isoform1 (SEQIDNO:188) mttsldtvetfgttsyyddvgllcekadtralmaqfvppl yslvftvgllgnvvvymilikyrrlrimtniyllnlaisd llflvtlpfwihyvrghnwvfghgmckllsgfyhtglyse iffiilltidrylaivhavfalrartvtfgvitsivtwgl avlaalpefifyeteelfeetlcsalypedtvyswrhfht lrmtifclvlpllvmaicytgiiktllrcpskkkykairl ifvimavffifwtpynvaillssyqsilfgndcerskhld lvmlvteviayshccmnpviyafvgerfrkylrhffhrhl lmhlgryipflpseklertssvspstaepelsivf HumanCCR3Isoform2 (SEQIDNO:189) mpfgirmllrahkpgssrrsemttsldtvetfgttsyydd vgllcekadtralmaqfvpplyslvftvgllgnvvvvmil ikyrrlrimtniyllnlaisdllflvtlpfwihyvrghnw vfghgmckllsgfyhtglyseiffiilltidrylaivhav falrartvtfgvitsivtwglavlaalpefifyeteelfe etlcsalypedtvyswrhfhtlrmtifclvlpllvmaicy tgiiktllrcpskkkykairlifvimavffifwtpynvai llssyqsilfgndcerskhldlvmlvteviayshccmnpv iyafvgerfrkylrhffhrhllmhlgryipflpseklert ssvspstaepelsivf HumanCCR3Isoform3 (SEQIDNO:190) mpfgirmllrahkpgrsemttsldtvetfgttsyyddvgl lcekadtralmaqfvpplyslvftvgllgnvvvvmiliky rrlrimtniyllnlaisdllflvtlpfwihyvrghnwvfg hgmckllsgfyhtglyseiffiilltidrylaivhavfal rartvtigvitsivtwglavlaalpefifyeteeffeetl csalypedtvyswrhfhtlrmtifclvlpllvmaicytgi iktllrcpskkkykairlifvimavffifwtpynvaills syqsilfgndcerskhldlvmlvteviayshccmnpviya fvgerfrkylrhffhrhllmhlgryipflpseklertssv spstaepelsivf HumanCCR5 (SEQIDNO:191) mdyqvsspiydinyytsepcqkinvkqiaarllpplyslv fifgfvgnmlvililinckrlksmtdiyllnlaisdlffl ltvpfwahyaaaqwdfgntmcqlltglyfigffsgiffii lltidrylavvhavfalkartvtfgvvtsvitwvvavfas lpgiiftrsqkeglhytcsshfpysqyqfwknfqtlkivi lglvlpllvmvicysgilktllrcrnekkrhravrlifti mivyflfwapynivlllntfqeffglnncsssnrldqamq vtetlgmthccinpiiyafvgekfrnyllvffqkhiakrf ckccsifqqeaperassvytrstgeqeisvgl

CCL11 InhibitorsAntibodies

[0868] In some embodiments, the CCL11 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCL11, CCR2, CCR3, or CCR5, or can specifically bind to two or more of CCL11, CCR2, CCR3, and CCR5. In some embodiments, a CCL11 inhibitor can bind to two or more of CCR2, CCR3, and CCR5.

[0869] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; and Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0870] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0871] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; and Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0872] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; and Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Pat. Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895; US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US 2017/0002082, each of which is incorporated by reference in its entirety.

[0873] In some examples the chemokine/chemokine receptor inhibitor is bertilimumab (Immune Pharmaceuticals), an anti-eotaxin-1 monoclonal antibody that targets CCL11, and is currently in a Phase II clinical study for ulcerative colitis. Additional examples of CCL11 inhibitors are described in U.S. Patent Application Publication Nos. 2016/0289329, 2015/0086546, 2014/0342450, 2014/0178367, 2013/0344070, 2013/0071381, 2011/0274696, 2011/0038871, 2010/0074886, 2009/0297502, 2009/0191192, 2009/0169541, 2009/0142339, 2008/0268536, 2008/0241923, 2008/0241136, 2005/0260139, 2005/0048052, 2004/0265303, 2004/0132980, 2004/0126851, 2003/0165494, 2002/0150576, 2002/0150570, 2002/0051782, 2002/0051781, 2002/0037285, 2002/0028436, 2002/0015700, 2002/0012664, 2017/0131282, 2016/0368979, 2016/0208011, 2011/0268723, 2009/0123375, 2007/0190055, 2017/0049884, 2011/0165182, 2009/0226434, 2009/0110686, 2009/0047735, 2009/0028881, 2008/0107647, 2008/0107595, 2008/0015348, 2007/0274986, 2007/0231327, 2007/0036796, 2007/0031408, 2006/0229336, 2003/0228306, 2003/0166870, 2003/0003440, 2002/0019345, and 2001/0000241, each of which is incorporated by reference in its entirety (e.g., the description of CCL11 inhibitors).

[0874] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0875] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0876] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0877] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0878] Additional examples of CCL11 inhibitors that are antibodies or antigen-binding antibody fragments are known in the art.

CXCL10 InhibitorsSmall Molecules and Peptides

[0879] In some instances, the CXCL10 inhibitor is a small molecule. For example, the CXCL10 inhibitor can be ganodermycin (see, e.g., Jung et al., J. Antiobiotics 64:683-686, 2011). Additional exemplary small molecule CXCL10 inhibitors are described in: U.S. Patent Application Publication No. 2005/0075333; U.S. Patent Application Publication No. 2004/0242498; U.S. Patent Application Publication No. 2003/0069234; U.S. Patent Application Publication No. 2003/0055054; U.S. Patent Application Publication No. 2002/0169159; WO 97/24325; WO 98/38167; WO 97/44329; WO 98/04554; WO 98/27815; WO 98/25604; WO 98/25605; WO 98/25617; WO 98/31364; Hesselgesser et al., J. Biol. Chem. 273(25):15687-15692 (1998); and Howard et al., J. Med. Chem. 41(13):2184-2193 (1998).

[0880] In some examples, the CXCL10 inhibitor is a peptide antagonist of a CXCR3 receptor (e.g., as described in U.S. Patent Application Publication No. 2007/0116669, 2006/0204498, and WO 98/09642). In some examples, the CXCL10 inhibitor is a chemokine mutant or analogue, e.g., those described in U.S. Pat. No. 5,739,103, WO 96/38559, and WO 98/06751. Additional examples of CXCL10 inhibitors that are small molecules or peptides are known in the art.

CCR2 Inhibitors

[0881] As used herein CCR2, CC chemokine receptor 2, or MCP-1 can be used interchangeably. CCL2, CCL8, and CCL16 each individually bind to CCR2.

[0882] The term CCR2 inhibitor refers to an agent which decreases the ability of CCR2 to bind to one or more (e.g., two, or three) of CCL2, CCL8, and CCL16.

[0883] In some embodiments, the CCR2 inhibitor can decrease the binding between CCL2 and CCR2 by blocking the ability of CCL2 to interact with CCR2. In some embodiments, the CCR2 inhibitor can decrease the binding between CCL8 and CCR2 by blocking the ability of CCL8 to interact with CCR2. In some embodiments, the CCR2 inhibitor can decrease the binding between CCL16 and CCR2 by blocking the ability of CCL16 to interact with CCR2.

[0884] In some embodiments, the CCR2 inhibitor decreases the ability of CCR2 to bind to each of CCL2 and CCL8. In some embodiments, the CCR2 inhibitor decreases the ability of CCR2 to bind to each of CCL2 and CCL16. In some embodiments, the CCR2 inhibitor decreases the ability of CCR2 to bind to each of CCL8 and CCL16. In some embodiments, the CCRS inhibitor decreases the ability of CCR2 to bind to each of CCL2, CCL8, and CCL16.

[0885] In some instances, the CCR2 inhibitor is a small molecule. In some instances, the CCR2 inhibitor is an antibody or an antigen-binding antibody fragment. In some instances, the CCR2 inhibitor is a peptide.

[0886] Exemplary sequences for human CCR2, human CCL2, human CCL8, and human CCL16 are shown below.

TABLE-US-00021 HumanCCR2IsoformA (SEQIDNO:192) mlstsrsrfirntnesgeevttffdydygapchkfdvkqi gaqllpplyslvfifgfvgnmlvvlilinckklkcltdiy llnlaisdllflitlplwahsaanewvfgnamcklftgly higyfggiffiilltidrylaivhavfalkartvtfgvvt svitwlvavfasvpgiiftkcqkedsvyvcgpyfprgwnn fhtimrnilglvlpllimvicysgilktllrcrnekkrhr avrviftimivyflfwtpynivillntfqeffglsncest sqldqatqvtetlgmthccinpiiyafvgekfrslfhial gcriaplqkpvcggpgvrpgknvkvttqglldgrgkgksi grapeaslqdkega HumanCCL2IsoformB (SEQIDNO:193) mlstsrsrfirntnesgeevttffdydygapchkfdvkqi gaqllpplyslvfifgfvgnmlvvlilinckklkcltdiy llnlaisdllflitlplwahsaanewvfgnamcklftgly higyfggiffiilltidrylaivhavfalkartvtfgvvt svitwlvavfasvpgiiftkcqkedsvyvcgpyfprgwnn fhtimrnilglvlpilimvicysgilktllrcrnekkrhr avrviftimivyflfwtpynivillntfqeffglsncest sqldqatqvtetlgmthccinpiiyafvgekfrrylsvff rkhitkrfckqcpvfyretvdgvtstntpstgeqevsagl HumanCCL8 (SEQIDNO:194) qpdsvsipitccfnvinrkipiqrlesytritniqcp keavifktkrgkevcadpkerwvrdsmkhldqifqnlkp HumanCCL16 (SEQIDNO:195) qpkvpewvntpstcclkyyekvlprrlvvgyrkalnc hlpaiifvtkrnrevanpnddwvqeyikdpnlpllptrn lstvkiitakngqpqllnsq

CCR2 InhibitorsAntibodies

[0887] In some embodiments, the CCR2 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCR2. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCL2. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCL8. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCL16. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCR2 and one or more of (e.g., one, two, or three) of CCL2, CCL8, and CCL16.

[0888] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; and Hudson et al., J Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; and Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0889] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0890] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; and Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0891] In some embodiments, the CCR2 inhibitor is a monoclonal antibody. For example, the CCR2 inhibitor can be MLN1202 (Millennium Pharmaceuticals), C775, STI-B0201, STI-B0211, STI-B0221, STI-B0232, carlumab (CNTO 888; Centocor, Inc.), or STI-B0234, or an antigen-binding fragment thereof. See also, e.g., Vergunst et al., Arthritis Rheum. 58(7):1931-1939, 2008. Additional examples of CCR2 inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., U.S. Patent Application Publication Nos. 2015/0086546, 2016/0272702, 2016/0289329, 2016/0083482, 2015/0361167; 2014/0342450, 2014/0178367, 2013/0344070, 2013/0071381, 2011/0274696, 2011/0059107, 2011/0038871, 2009/0068109, 2009/0297502, 2009/0142339, 2008/0268536, 2008/0241923, 2008/0241136, 2007/0128112, 2007/0116708, 2007/0111259, 2006/0246069, 2006/0039913, 2005/0232923, 2005/0260139, 2005/0058639, 2004/0265303, 2004/0132980, 2004/0126851, 2004/0219644, 2004/0047860, 2003/0165494, 2003/0211105, 2002/0150576, 2002/0051782, 2002/0042370, and 2002/0015700; and U.S. Pat. Nos. 6,312,689, 6,084,075, 6,406,694, 6,406,865, 6,696,550, 6,727,349, 7,442,775, 7,858,318, 5,859,205, 5,693,762, and 6,075,181, each of which is incorporated by reference (e.g., the description of the CCR2 inhibitors). Additional examples of CCR2 inhibitors are described in, e.g., WO 00/05265. Additional examples of CCR2 inhibitors that are antibodies or antigen-binding antibodies fragments are described in, e.g., Loberg et al., Cancer Res. 67(19):9417, 2007.

[0892] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0893] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0894] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0895] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0896] Additional examples of CCR2 inhibitors that are antibodies or antigen-binding antibody fragments are known in the art.

CCR2 InhibitorsSmall Molecules and Peptides

[0897] In some examples, the CCR2 inhibitor is a small molecule. For example, the CCR2 inhibitor can be elubrixin, PF-04634817, BMS-741672, or CCX872. See, e.g., U.S. Pat. No. 9,434,766; U.S. Patent Application Publication No. 20070021466; Deerberg et al., Org. Process Rev. Dev. 20(11):1949-1966, 2016; and Morganti et al., J. Neurosci. 35(2):748-760, 2015.

[0898] Additional non-limiting examples of CCR2 inhibitors that are small molecules include, e.g., the phenylamino substituted quaternary salt compounds described in U.S. Patent Application Publication No. 2009/0112004; the biaryl derivatives described in U.S. Patent Application Publication No. 2009/0048238; the pyrazol derivatives described in U.S. Patent Application Publication No. 2009/0029963; the heterocyclic compounds described in U.S. Patent Application Publication No. 2009/0023713; the imidazole derivatives described in U.S. Patent Application Publication No. 2009/0012063; the aminopyrrolidines described in U.S. Patent Application Publication No. 2008/0176883; the heterocyclic cyclopentyl tetrahydroisoquinolones and tetrahydropyridopyridines described in U.S. Patent Application Publication No. 2008/0081803; the heteroaryl sulfonamides described in U.S. Patent Application Publication No. 2010/0056509; the triazolyl pyridyl benzenesulfonamides described in U.S. Patent Application Publication No. 2010/0152186; the bicyclic and bridged nitrogen heterocycles described in U.S. Patent Application Publication No. 2006/0074121; the fused heteroaryl pyridyl and phenyl benzenesulfonamides described in WO 09/009740; and the 3-aminopyrrolidene derivatives described in WO 04/050024.

[0899] Additional non-limiting examples of CCR2 inhibitors include: N-((1R,3S)-3-isopropyl-3-{[3-(trifluoromethyl)-7,8-dihydro-1,6-naph-thyri-din-6(5H)-yl]carbonyl}cyclopentyl)-N-[(3S,4S)-3-methoxytetrahydro-2H-pyran- -4-yl]amine; 3[(3S,4R)-1-((1R,3S)-3-isopropyl-2-oxo-3-{[6-(trifluoromethyl)-2H-1,3-ben-z-oxazin-3(4H)-yl]methyl}cyclopentyl)-3-methylpiperidin-4-yl]benzoic acid; (3S,48)-N-((1R,3S)-3-isopropyl-3-{[7-(trifluoromethyl)-3,4-dihydroisoquin-olin-2(1B)-yl]carbonyl}cyclopentyl)-3-methyltetrahydro-2H-p-yran-4-aminium; 3-[(3S,4R or 3R,4S)-1-((1R,3S)-3-Isopropyl-3-{[6-(trifluoromethyl)-2H-1,3-benzoxazin-3-(4H)-yl]carbonyl}cyclopentyl)-3-methylpiperidin-4-yl]benzoic acid; INCB3284; Eotaxin-3; PF-04178903 (Pfizer), and pharmaceutically acceptable salts thereof.

[0900] Additional non-limiting examples of CCR2 inhibitors include: bindarit (2-((1-benzyl-1H-indazol-3-yl)methoxy)-2-methylpropionic acid); AZD2423 (AstraZeneca); the indole describes described in U.S. Pat. Nos. 7,297,696, 6,962,926, 6,737,435, and 6,569,888; the bicyclic pyrrole derivatives described in U.S. Pat. Nos. 6,441,004 and 6,479,527; the CCR2 inhibitors described in U.S. Patent Application Publications Nos. 2005/0054668, 2005/0026975, 2004/0198719, and 2004/0047860, and Howard et al., Expert Opin. Ther. Patents 11(7):1147-1151 (2001).

[0901] Additional non-limiting examples of CCR2 inhibitors that are small molecules are described in, e.g., WO 97/24325; WO 98/38167; WO 97/44329; WO 98/04554; WO 98/27815; WO 98/25604; WO 98/25605; WO 98/25617; WO 98/31364; Hesselgesser et al., J. Biol. Chem. 273(25):15687-15692, 1998; and Howard et al., J. Med. Chem. 41(13):2184-2193, 1998.

[0902] In some embodiments, the CCR2 inhibitor is a small nucleic acid, e.g., NOX-E36 (a 40-nucleotide L-RNA oligonucleotide that is linked to a 40-kDa PEG; NOXXON Pharma AG).

[0903] In some embodiments, the CCR2 inhibitor is a peptide, e.g., a dominant negative peptide described in, e.g., Kiyota et al., Mol. Ther. 17(5):803-809, 2009, and U.S. Patent Application Publication No. 20070004906, or an antagonistic peptide, e.g., the antagonistic peptides described in WO 05/037305 and Jiang-Hong Gong, et al., J. Exp. Med. 186:131, 1997. Additional examples of CCR2 inhibitors that are peptides are described in, e.g., U.S. Pat. No. 5,739,103; WO 96/38559; WO 98/06751; and WO 98/09642. In some embodiments, a CCR2 inhibitor is a CCR2 mutein (e.g., U.S. Patent Application Publication No. 2004/0185450).

[0904] Additional examples of CCR2 inhibitors that are small molecules and peptides are known in the art.

CCR9 Inhibitors

[0905] As used herein CCR9 or CC chemokine receptor 9 can be used interchangeably. CCR9 specifically binds to CCL25.

[0906] The term CCR9 inhibitor refers to an agent which decreases the ability of CCR9 to bind to CCL25.

[0907] In some embodiments, the CCR9 inhibitor can decrease the binding between CCL25 and CCR9 by blocking the ability of CCL25 to interact with CCR9. In some instances, the CCR9 inhibitor is a small molecule. In some instances, the CCR9 inhibitor is an antibody or an antigen-binding antibody fragment.

[0908] Exemplary sequences for human CCR9 and CCL25 are shown below.

TABLE-US-00022 HumanCCR9IsoformA (SEQIDNO:196) mtptdftspipnmaddygsestssmedyvnfnftdfycek nnvrqfashflpplywlvfivgalgnslvilvywyctrvk tmtdmfllnlaiadllflvtlpfwaiaaadqwkfqtfmck vvnsmykmnfyscvllimcisvdryiaiaqamrahtwrek rllyskmvcftiwvlaaalcipeilysqikeesgiaictm vypsdestklksavltlkvilgfflpfvvmaccytiiiht liqakksskhkalkvtitvltvfvlsqfpyncillvqtid ayamfisncavstnidicfqvtqtiaffhsclnpvlyvfv gerfrrdlvktlknlgcisqaqwvsftrregslklssmll ettsgalsl HumanCCR9IsoformB (SEQIDNO:197) maddygsestssmedyvnfnftdfyceknnvrqfashflp plywlvfivgalgnslvilvywyctrvktmtdmfllnlai adllflvtlpfwaiaaadqwkfqtfmckvvnsmykmnfys cvllimcisvdryiaiaqamrahtwrekrllyskmvcfti wvlaaalcipeilysqikeesgiaictmvypsdestklks avltlkvilgfflpfvvmaccytiiihtliqakksskhka lkvtitvltvfvlsqfpyncillvqtidayamfisncavs tnidicfqvtqtiaffhsclnpvlyvfvgerfrrdlyktl knlgcisqaqwvsftrregslklssmllettsgalsl HumanCCL25Isoform1 (SEQIDNO:198) qgvfedcclayhypigwavlrrawtyriqevsgscnl paaifylpkrhrkvcgnpksrevqramklldarnkvfakl hhntqtfqagphavkklssgnsklssskfsnpissskrnv sllisansgl HumanCCL25Isoform2 (SEQIDNO:199) qgvfedcclayhypigwavlrrawtyriqevsgscnl paaifylpkrhrkvcgnpksrevqramklldarnkvfakl hhntqtfqgphavkklssgnsklssskfsnpissskrnvs llisansgl

CCR9 InhibitorsAntibodies

[0909] In some embodiments, the CCR9 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCR9. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CCL25. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to both CCR9 and CCL25.

[0910] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; and WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; and Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther 8(3):355-366, 2003; and Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0911] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0912] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; and Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0913] In other instances, the CCR9 inhibitor is a monoclonal antibody. For example, the CCR9 antibody can be 91R, see, e.g., Chamorro et al., MAbs 6(4): 1000-1012, 2014. Additional non-limiting examples of CCR9 inhibitors are described in, e.g., U.S. Patent Application Publication Nos. 2012/0100554, 2012/0100154, 2011/0123603, 2009/0028866, and 2005/0181501.

[0914] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0915] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510 8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110-M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0916] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0917] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0918] Additional examples of CCR9 inhibitors that are antibodies or antigen-binding antibody fragments are known in the art.

CCR9 InhibitorsSmall Molecules

[0919] In some instances, the CCR9 inhibitor is a small molecule. For example, the CCR9 inhibitor can be Traficet-EN (also called Vercirnon, CCX282, and GSK1605786) or Tu1652 CCX507. See, e.g., Eksteen et al., IDrugs 13(7):472-481, 2010; and Walters et al., Gastroenterology 144(5):S-815, 2013.

[0920] Additional examples of CCR9 inhibitors that are small molecules are known in the art.

ELR Chemokine Inhibitors

[0921] ELR chemokines are CXC chemokines that have a glutamic acid-leucine-arginine (ELR) motif. See, e.g., Strieter et al., J Biol. Chem. 270:27348-27357, 1995.

[0922] The term ELR chemokine inhibitor refers to an agent which decreases the ability of CXCR1 and/or CXCR2 to bind to one or more (e.g., two, three, four, five, six, seven, or eight) of CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, and CXCL8.

[0923] In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR1 and CXCL8 by blocking the ability of CXCR1 to interact with CXCL8. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR1 and CXCL6 by blocking the ability of CXCR1 to interact with CXCL6. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR1 and each of CXCL8 and CXCL6.

[0924] In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL1 by blocking the ability of CXCR2 to interact with CXCL1. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL2 by blocking the ability of CXCR2 to interact with CXCL2. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL3 by blocking the ability of CXCR2 to interact with CXCL3. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL4 by blocking the ability of CXCR2 to interact with CXCL4. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL5 by blocking the ability of CXCR2 to interact with CXCL5. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL6 by blocking the ability of CXCR2 to interact with CXCL6. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and CXCL7 by blocking the ability of CXCR2 to interact with CXCL7. In some embodiments, the ELR chemokine inhibitor can decrease the binding between CXCR2 and one or more (e.g., two, three, four, five, six, or seven) of CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, and CXCL7.

[0925] In some embodiments, the ELR chemokine inhibitor can decrease the binding of CXCR1 to one or both of CXCL6 and CXCL8, and can decrease the binding to CXCR2 to one or more (e.g., two, three, four, five, six, or seven) of CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, and CXCL7

[0926] In some instances, the ELR chemokine inhibitor is a small molecule. In some instances, the ELR chemokine inhibitor is an antibody or an antigen-binding antibody fragment.

[0927] Exemplary sequences for human CXCR1, human CXCR2, human CXCL1, human CXCL2, human CXCL3, human CXCL4, human CXCL5, human CXCL6, human CXCL7, and human CXCL8.

TABLE-US-00023 HumanCXCR1 (SEQIDNO:200) msnitdpqmwdfddlnftgmppadedyspcxletetlnky vviiayalvfllsllgnslvmlvilysrvgrsvtdvylln laladllfaltlpiwaaskvngwifgtflckvvsllkevn fysgilllacisvdrylaivhatrtltqkrhlvkfvclgc wglsmnlslpfflfrqayhpnnsspvcyevlgndtakwrm vlrilphtfgfivplfvmlfcygftlrtlfkahmgqkhra mrvifavvlifllcwlpynlvlladtlmrtqviqescerr nnigraldateilgflhsclnpiiyafigqnfrhgflkil amhglvskeflarhrvtsytsssvnvssnl HumanCXCR2 (SEQIDNO:201) medfnmesdsfedfwkgedlsnysysstlppflldaapce pesleinkyfvviiyalvfllsllgnslvmlvilysrvgr svtdvyllnlaladllfaltlpiwaaskvngwifgtflck vvsllkevnfysgilllacisvdrylaivhatrtltqkry lvkficlsiwglslllalpvllfrrtvyssnvspacyedm gnntanwrmllrilpqsfgfivpllimlfcygftlrtlfk ahmgqkhramrvifavvlifllcwlpynlvlladtlmrtq viqetcerrnhidraldateilgilhsclnpliyafigqk frhgllkilaihgliskdslpkdsrpsfvgsssghtsttl HumanCXCL1 (SEQIDNO:202) maraalsaapsnprllrvallllllvaagrraagasvate lrcqclqtlqgihpkniqsvnvkspgphcaqteviatlkn grkaclnpaspivkkiiekmlnsdksn HumanCXCL2 (SEQIDNO:203) maratlsaapsnprllrvallllllvaasrraagaplate lrcqclqtlqgihlkniqsvkvkspgphcaqteviatlkn gqkaclnpaspmvkkiiekmlkngksn HumanCXCL3 (SEQIDNO:204) asvvtelrcqclqtlqgihlkniqsvnvrspgphca qteviatlkngkkaclnpaspmvqkiiekilnkgstn HumanCXCL4 (SEQIDNO:205) mssaagfcasrpgllflgllllplvvafasaeaeedgdlq clcvkttsqvrprhitslevikagphcptaqliatlkngr kicldlqaplykkiikklles HumanCXCL5 (SEQIDNO:206) msllssraarvpgpssslcallvllllltqpgpiasagpa aavlrelrcvclqttqgvhpkmisnlqvfaigpqcskvev vaslkngkeicldpeapflkkviqkildggnken HumanCXCL6 (SEQIDNO:207) gpvsavltelrctclrvtlrvnpktigklqvfpagpqcskvev vaslkngkqvcldpeapflkkviqkildsgnkkn HumanCXCL7 (SEQIDNO:208) mslrldttpscnsarplhalqvllllsllltalasstkgq tkrnlakgkeesldsdlyaelrcmcikttsgihpkniqsl evigkgthcnqveviatlkdgrkicldpdaprikkivqkk lagdesad HumanCXCL8Isoform1 (SEQIDNO:209) egavlprsakelrcqciktyskpfhpkfikelrviesgph canteiivklsdgrelcldpkenwvqrvvekflkraens HumanCXCL8Isoform2 (SEQIDNO:210) egavlprsakelrcqciktyskpfhpkfikelrviesgph canteiivklsdgrelcldpkenwvqrvvekflkr

ELR Chemokine InhibitorsAntibodies

[0928] In some embodiments, the ELR chemokine inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CXCR1 and/or CXCR2. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to one or more (e.g., two, three, four, five, six, seven, or eight) of: CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, and CXCL8 (IL-8).

[0929] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; and WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; and Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther 8(3):355-366, 2003; and Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[0930] Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab).sub.2 fragment, and a Fab fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

[0931] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; and Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[0932] An ELR chemokine inhibitor can be, e.g., a monoclonal antibody. A non-limiting example of an ELR inhibitor is TAB-099MZ. Additional examples of ELR chemokine inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., U.S. Pat. No. 9,290,570; and U.S. Patent Application Publication Nos. 2004/0170628, 2010/0136031, 2015/0160227, 2015/0224190, 2016/0060347, 2016/0152699, 2016/0108117, 2017/0131282, 2016/0060347, 2014/0271647, 2014/0170156, 2012/0164143, 2010/0254941, 2009/0130110, 2008/0118517, 2004/0208873, 2003/0021790, 2002/0082396, and 2001/0006637, each of which is herein incorporated by reference (e.g., the portions describing ELR chemokine inhibitors).

[0933] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (K.sub.D) of less than 110.sup.5 M (e.g., less than 0.510.sup.5 M, less than 110.sup.6 M, less than 0.510.sup.6 M, less than 110.sup.7 M, less than 0.510.sup.7 M, less than 110.sup.8 M, less than 0.510.sup.8 M, less than 110.sup.9 M, less than 0.510.sup.9 M, less than 110.sup.10 M, less than 0.510.sup.10 M, less than 110.sup.11 M, less than 0.510.sup.11 M, or less than 110.sup.12 M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0934] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.D of about 110.sup.12 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510 8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, about 110.sup.11 M, or about 0.510.sup.11 M (inclusive); about 0.510.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, about 0.510.sup.10 M, or about 110.sup.11 M (inclusive); about 110.sup.11 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, about 110.sup.10 M, or about 0.510.sup.10 M (inclusive); about 0.510.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, about 0.510.sup.9 M, or about 110.sup.10 M (inclusive); about 110.sup.10 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, about 110.sup.9 M, or about 0.510.sup.9 M (inclusive); about 0.510.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, about 0.510.sup.8 M, or about 110.sup.9 M (inclusive); about 110.sup.9 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, about 110.sup.8 M, or about 0.510.sup.8 M (inclusive); about 0.510.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, about 0.510.sup.7 M, or about 110.sup.8 M (inclusive); about 110.sup.8 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, about 110.sup.7 M, or about 0.510.sup.7 M (inclusive); about 0.510.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, about 0.510.sup.6 M, or about 110.sup.7 M (inclusive); about 110.sup.7 M to about 110.sup.5 M, about 0.510.sup.5 M, about 110.sup.6 M, or about 0.510.sup.6 M (inclusive); about 0.510.sup.6 M to about 110.sup.5 M, about 0.510.sup.5 M, or about 110.sup.6 M (inclusive); about 110.sup.6 M to about 110.sup.5 M or about 0.510.sup.5 M (inclusive); or about 0.510.sup.5 M to about 110.sup.5 M (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0935] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.off of about 110.sup.6 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, about 110.sup.5 s.sup.1, or about 0.510.sup.5 s.sup.1 (inclusive); about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, about 0.510.sup.4 s.sup.1, or about 110.sup.5 s.sup.1 (inclusive); about 110.sup.5 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, about 110.sup.4 s.sup.1, or about 0.510.sup.4 s.sup.1 (inclusive); about 0.510.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, about 0.510.sup.3 s.sup.1, or about 110.sup.4 s.sup.1 (inclusive); about 110.sup.4 s.sup.1 to about 110.sup.3 s.sup.1, or about 0.510.sup.3 s.sup.1 (inclusive); or about 0.510.sup.5 s.sup.1 to about 110.sup.3 s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0936] In some embodiments, any of the antibodies or antigen-binding fragments described herein has a K.sub.on of about 110.sup.2 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, about 110.sup.3 M.sup.1s.sup.1, or about 0.510.sup.3 M.sup.1s.sup.1 (inclusive); about 0.510.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, about 0.510.sup.4 M.sup.1s.sup.1, or about 110.sup.3 M.sup.1s.sup.1 (inclusive); about 110.sup.3 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, about 110.sup.4 M.sup.1s.sup.1, or about 0.510.sup.4 M.sup.1s.sup.1 (inclusive); about 0.510.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, about 0.510.sup.5 M.sup.1s.sup.1, or about 110.sup.4 M.sup.1s.sup.1 (inclusive); about 110.sup.4 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, about 110.sup.5 M.sup.1s.sup.1, or about 0.510.sup.5 M.sup.1s.sup.1 (inclusive); about 0.510.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, about 0.510.sup.6 M.sup.1s.sup.1, or about 110.sup.5 M.sup.1s.sup.1 (inclusive); about 110.sup.5 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1, or about 0.510.sup.6 M.sup.1s.sup.1 (inclusive); or about 0.510.sup.6 M.sup.1s.sup.1 to about 110.sup.6 M.sup.1s.sup.1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

[0937] Additional examples of ELR chemokine inhibitors that are antibodies or antigen-binding antibody fragments are known in the art.

ELR Chemokine InhibitorsSmall Molecules

[0938] In some instances, the ELR chemokine inhibitor is, e.g., a small molecule. For example, the ELR chemokine inhibitor can be, e.g., LY-3041658 or repertaxin (Reparixin; DF 1681Y). Additional non-limiting examples of ELR chemokine inhibitors that are small molecules are described in, e.g., U.S. Patent Application Publication Nos. 2007/0248594, 2006/0014794, 2004/0063709, 2004/0034229, 2003/0204085, 2003/0097004, 2004/0186142, 2004/0235908, 2006/0025453, 2017/0224679, 2017/0190681, 2017/0144996, and 2017/0128474, each of which are incorporated by reference (e.g., the portions describing the ELR chemokine inhibitors).

[0939] In some embodiments, the ELR chemokine inhibitor is a peptide, e.g., any of the peptides described in U.S. Patent Application Publication Nos. 2009/0270318, 2009/0118469, and 2007/0160574, 2007/0021593, 2003/0077705, and 2007/0181987, each of which is incorporated by reference (e.g., the portions describing the ELR chemokine inhibitors).

[0940] Phosphodiesterase 4 (PDE4) Inhibitors

[0941] The term PDE4 inhibitor refers to an agent which decreases PDE4 activity in vitro or in a mammalian cell, e.g., as compared to the level of PDE4 activity in the absence of the agent; and/or decreases the level of a PDE4 protein in a mammalian cell contacted with the agent, e.g., as compared to the same mammalian cell not contacted with the agent. A non-limiting example of PDE4 activity is the degradation of cAMP.

[0942] In some embodiments, a PDE4 inhibitor can be a small molecule (e.g., an organic, an inorganic, or bioinorganic molecule) having a molecule weight of less than 900 Daltons (e.g., less than 500 Daltons). In some embodiments, a PDE4 inhibitor can be an inhibitory nucleic acid.

Small Molecules

[0943] In some embodiments, a PDE4 inhibitor is a small molecule. Non-limiting examples of small molecules that are PDE4 inhibitors are shown in Table A.

[0944] Table A. Exemplary Small Molecules that are PDE4 Inhibitors

TABLE-US-00024 TABLE A Exemplary Small Molecules that are PDE4 Inhibitors Originator Other Drug Drug Name Company Structure Names Indications apremilast Celgene Corp [00018] CC-10004; CC-110004; CDC-104; Otezla; apremilast; lead seICID (2), Celgene seICID (COPD), Celgene; Asthma; Atopic dermatitis; Crohns disease; Inflammatory disease; Rheumatoid arthritis CC-1088 Celgene Corp [00019] CC-1088; Corp CC-5048; CC-801; CDC-801; lead SeICID (1), Celgene Crohns disease; Inflammatory disease; Myelodysplastic syndrome tetomilast Otsuka Pharmaceutical Co Ltd [00020] OPC-6535; tetomilast Chronic obstructive pulmonary disease; Crohns disease; Inflammatory bowel disease; Respiratory disease; Ulcerative colitis KF-19514 Kyowa Hakko Kogyo Co Ltd [00021] KF-19514; PDE 4 inhibitors (asthma), Kyowa Allergic rhinitis; Asthma; Respiratory disease PF-06266047 Pfizer Inc PF-06266047 Schizophrenia SKF-107806 SmithKline SKF-107806 Asthma Beecham plc PDB-093 Wyeth- Ayerst Pharmaceuticals Inc [00022] PDB-093 Asthma PDE4 inhibitors (inhalant formulation, chronic obstructive pulmonary disease), AstraZeneca AstraZeneca plc [00023] PDE4 inhibitors (inhalant formulation, chronic obstructive pulmonary disease), AstraZeneca Chronic obstructive pulmonary disease tolafentrine Takeda GmbH [00024] BY-4070; tolafentrine Asthma TAK-648 Takeda TAK-648 Diabetic Pharmaceutical nephropathy; Co Ltd Non-insulin dependent diabetes CH-928 UCB CH-928 Asthma Celltech CH-673 UCB CH-673 Asthma Celltech CH-422 UCB CH-422 Asthma Celltech ABI-4 Pfizer Inc [00025] 18F-PF- 06445974; ABI-4; Fluorine-18-PF- 06445974; PDE4 inhibitor (psychotic disorders), Pfizer/ Northeastern University; PF-06445974- -[18F] roflumilast N- oxide (inhalant formulation, airway disorders), Incozen Incozen Therapeutics Pvt Ltd [00026] roflumilast N-oxide; roflumilast N-oxide (inhalant formulation, airway disorders), Inconzen Respiratory disease PDE4 allosteric inhibitors (mild cognitive impairment/ traumatic brain injury), Tetra Discovery Tetra Discovery Partners LLC PDE4 allosteric inhibitors (mild cognitive impairment/ traumatic brain injury), Tetra Discovery PDE4 inhibitors (inflammatory disorders), Kyorin Pharmaceuticals Kyorin Pharmaceutical Co Ltd [00027] PDE4 inhibitors (inflammatory disorders), Kyorin Pharmaceuticals Inflammatory disease BYK-321084 Takeda BYK-321084 Psoriasis Pharma A/S WAY-127093B Wyeth- Ayerst Pharmaceuticals Inc [00028] WAY-127093B Asthma NCS-613 Centre National de la Recherche Scientifique (CNRS) [00029] NCS-613 Cardiac failure SDZ-ISQ-844 Novartis AG [00030] SDZ-ISQ-844 Asthma dual long-acting beta2- adrenoceptor agonists/PDE4 inhibitors (inhalant, COPD), Gilead Gilead Sciences Inc [00031] GS-5759; dual long-acting beta2-adrenoceptor agonists/PDE4 inhibitors (inhalant, COPD), Gilead Chronic obstructive pulmonary disease Ro-20-1724 Roche Holding AG [00032] Ro-20-1724 Asthma; Psoriasis Hemay-005 Tianjin Hemay-005; Hemay Bio- TNF alpha Tech Co Ltd and IL-1 dual antagonist (inflammation), Tianjin Hemay Bio- Tech/Hainan Hailing Chemipharma Corporation PDE3/PDE4 inhibitors, Kyorin Kyorin Holdings Inc [00033] KCA-1490; PDE3/PDE4 inhibitors, Kyorin Respiratory disease phospho- diesterase inhibitors, Syntex Roche Palo Alto [00034] PDE4 inhibitors, Syntex; TVX-2706; nitraquazone; phosphodiesterase inhibitors, Syntex Inflammatory disease filaminast Wyeth- Ayerst Pharmaceuticals Inc [00035] PDA-641; WAY-PDA-641; filaminast Asthma; Inflammatory disease LASSBio-596 LASSBio [00036] LASSBio-596; PDE4/PDE5 inhibitor (acute lung injury/ asthma), LASSBio Asthma ASP-3258 Astellas Pharma Inc [00037] ASP-3258; PDE 4 inhibitor (airway inflammation), Astellas; phosphodiesterase 4 inhibitor (airway inflammation), Astellas Respiratory tract inflammation TAS-203 Taiho Pharmaceutical Co Ltd [00038] PDE 4 inhibitor (airway inflammation), Taiho; TAS-203; phosphodiesterase 4 inhibitor (airway inflammation), Taiho Respiratory tract inflammation PDE4 inhibitor (inflammatory disease/ autoimmune disease), Anacor Pharmaceuticals Anacor Pharmaceuticals Inc [00039] AN-3889; AN-5322; AN-6414; AN-6415; PDE4 inhibitor (inflammatory disease/autoimmune disease), Anacor Pharmaceuticals lotamilast Eisai Co Ltd [00040] E-6005; RVT-501; lotamilast GPD-1116 ASKA Pharmaceutical Co Ltd [00041] GPD-1116 Asthma; Chronic obstructive pulmonary disease; Emphysema cipamfylline SmithKline Beecham plc [00042] BRL-61063; HEP-688; cipamfylline Asthma; Atopic dermatitis Phospho- Spring Bank Phosphodiesterase Chronic diesterase Pharmaceuticals 3, 4 and 7 obstructive 3, 4 and 7 Inc inhibitors (oral, pulmonary inhibitors (oral, COPD), Spring Bank disease COPD), Spring Pharmaceuticals; Bank SMNH compounds Pharmaceuticals (oral, COPD), Spring Bank Pharmaceuticals; (oral, COPD), Spring Bank Pharmaceuticals; nucleotide based program (oral, COPD), Spring Bank Pharmaceuticals; small molecule nucleic acid hybrids (oral, COPD), Spring Bank Pharmaceuticals ZL-N-91 Zhejiang ZL-N-91 Lung University inflammation PDE 4 inhibitors (inflammation), Almirall Almirall Prodesfarma SA [00043] PDE 4 inhibitors (inflammation), Almirall Inflammatory disease CDP-840 UCB Celltech [00044] CDP-840 Asthma GSK-356278 GlaxoSmith Kline plc [00045] 356278; GSK-356278; PDE4 inhibitor (oral, depression/anxiety, GlaxoSmithKline Anxiety disorder; Depression; Huntingtons chorea cilomilast SmithKline Beecham plc [00046] Ariflo; SB-207499; cilomilast; oral phosphodiesterase 4 inhibitor (asthma/COPD), GSK Asthma; Chronic obstructive pulmonary disease PDE4 inhibitors (oral, COPD), GlaxoSmithKline GlaxoSmith Kline plc [00047] PDE4 inhibitors (oral, COPD), GlaxoSmithKline Chronic obstructive pulmonary disease dual PDE4/ L-type calcium channel inhibitors (hypertension), University of South Carolina University of South Carolina [00048] MNP-001; MS-23; MSP-001; dual PDE4/L-type calcium channel inhibitors (hypertension), University of South Carolina Hypertension PDE-4 inhibitor Crystal PDE-4 inhibitor Asthma (asthma), Genomics (asthma), CrystalGenomics Inc CrystalGenomics PDE 4 inhibitors (dermatitis/ rheumatoid arthritis), Kyowa Hakko Kirin Kyowa Hakko Kirin Co Ltd [00049] K-34; KF-66490; PDE 4 inhibitors (dermatitis/ rheumatoid arthritis), Kyowa Hakko Kirin Atopic dermatitis; Rheumatoid arthritis cilomilast (ophthalmic disease), Alcon GlaxoSmith Kline plc [00050] AL-38583; cilomilast; cilomilast (ophthalmic disease), Alcon; cilomilast (ophthalmic disease), GSK Allergic conjunctivitis; Ocular disease; Xerophthalmia OCID-2987 Orchid OCID-2987; Asthma; Pharma Ltd PDE IV inhibitor Chronic (inflammation), obstructive Orchid; PDE4 pulmonary inhibitor disease; (inflammation), Inflammatory Orchid; disease phosphodiesterase IV inhibitor (inflammation), Orchid roflumilast (dermatolocrical, psoriasis/atopic dermatitis), Nycomed Takeda Pharmaceuticals International GmbH [00051] roflumilast; roflumilast (dermatological, psoriasis/atopic dermatitis), Nycomed Atopic dermatitis; Psoriasis PDE 4 inhibitor Takeda PDE 4 inhibitor Inflammatory (inflammation), Pharmaceuticals (inflammation), disease Takeda Takeda Pharmaceuticals International Pharmaceuticals International GmbH International AN-2898 Anacor Pharmaceuticals Inc [00052] AN-2898; PDE4 inhibitor (topical, psoriasis/atopic dermatitis), Anacor Atopic dermatitis; Psoriasis dual p38/PDE4 inhibitors (inflammation), c- a-i-r biosciences c-a-i-r biosciences GmbH [00053] CBS-3595; dual p38/PDE4 inhibitors (inflammation), c-a-i-r biosciences; dual p38/ phosphodiesterase 4 inhibitors (inflammation), c-a-i-r biosciences Inflammatory disease ASP-9831 Astellas Pharma Inc [00054] ASP-9831; PDE4 inhibitor (hepatitis), Astellas; PDE4 inhibitor (non-alcoholic steatohepatitis), Astellas Non-alcoholic steatohepatitis phospho- VIA phosphodiesterase Vasculitis diesterase Pharmaceuticals 4 inhibitors 4 inhibitors Inc (vascular (vascular inflammation), VIA inflammation), Pharmaceuticals VIA Pharmaceuticals E-4021 Eisai Co Ltd [00055] 4- Piperidinecarboxylic acid, 1-[4-[(1,3- benzodioxol-5- ylmethyl)amino]- 6-chloro-2- guinazolinyl]-; E-4021 Angina; Cardiac failure piclamilast Rhone- Poulenc SA [00056] RP-73401; RPR-73401; piclamilast Arthritis; Asthma CD-160130 Curacyte AG CD-160130; Chronic PDE-4 inhibitor lymphocytic (oral, B-CLL), leukemia BlackSwan Pharma; PDE-4 inhibitor (oral, B-CLL), Curacyte Discovery; PDE-4 inhibitor (oral, B-cell chronic lymphocytic leukemia), Curacyte Discovery GSK-256066 (allergic rhinitis, intranasal formulation), GlaxoSmithKline GlaxoSmith Kline plc [00057] 256066; 256066 (allergic rhinitis, intranasal formulation), GlaxoSmithKline; GSK-256066; GSK-256066 (allergic rhinitis, intranasal formulation), GlaxoSmithKline Allergic rhinitis 4AZA-PDE4 4 AZA 4AZA-PDE4 Immune Bioscience disorder NV YM-393059 Astellas Pharma Inc [00058] YM-393059; dual PDE7A/PDE4 inhibitors (immune disorder), Astellas Immune disorder revamilast Glenmark Pharmaceuticals Ltd [00059] GRC-4039; PDE 4 inhibitor (inflammation), Glenmark; phosphodiesterase 4 inhibitor (inflammation), Glenmark; revamilast; revamilast (inflammation), Glenmark Asthma; Inflammatory disease; Multiple sclerosis; Rheumatoid arthritis AN-2728 Anacor Pharmaceuticals Inc [00060] AN-2728; EUCRISA; EUCRISA; Eucrysa; Eucrysa; PF- 06930164; crisaborole MK-0952 Merck & Co Inc [00061] MK-0952; MK-952; PDE4 inhibitor (AD), Merck & Co; phosphodiesterase type 4 inhibitor (Alzheimer's disease), Merck & Co Alzheimer's disease ibudilast (oral, neuropathic pain/opiate dependence/ neuro- degeneration/T BI/druq dependence), Avigen Inc [00062] AV-411; MN-166; glial activation inhibitor (oral, neuropathic pain/opiate dependence), Avigen; ibudilast; ibudilast (oral, MediciNova neuropathic pain/opiate dependence/ alcohol dependence), Avigen; ibudilast (oral, neuropathic pain/opiate dependence/ neurodegeneration/ TBI/drug dependence), MediciNova; neurodegeneration disease therapy, Avigen; neuropathic pain therapy, Avigen GP-0203 Centre GP-0203; Asthma; National de la PDE 4 inhibitor Chronic Recherche (COPD/asthma), obstructive Scientifique CNRS pulmonary (CNRS) disease dual PDE 3/4 Scottish dual PDE 3/4 Asthma inhibitors (oral, Biomedical inhibitors (oral, asthma), Scottish Ltd asthma), Scottish Biomedical Biomedical; dual phospho- diesterase 3/4 inhibitors (oral, asthma), Scottish Biomedical ELB-526 elbion AG ELB-526; Lung inhaled PDE 4 inflammation inhibitor (lung inflammation), elbion; inhaled phosphodiesterase 4 inhibitor (lung inflammation), elbion theophylline (SODAS/Pharma Zome), Elan Elan Corp plc [00063] Teonova; Theolan; once-daily theophylline (SODAS), Elan; theophylline; theophylline (PharmaZome), Elan; theophylline (SODAS), Elan; theophylline (SODAS/ PharmaZome), Elan; theophylline, Elan; twice-daily theophylline (PharmaZone), Elan CHF-6001 Chiesi Farmaceutici SpA [00064] CHF-5480; CHF-6001; PDE 4 inhibitors (inhalant formulation, COPD/asthma), Chiesi elbimilast elbion AG [00065] AWD-12-353; ELB-353; PDE4 inhibitor, BioTie; PDE4 inhibitor, elbion; elbimilast; ronomilast AWD-12-281 (topical cream), elbion/ GlaxoSmithKline elbion AG [00066] 842470; AWD-12-281; AWD-12-281 (dermatitis), elbion/ GlaxoSmithKline; AWD-12-281 (topical cream), elbion/ GlaxoSmithKline; GW-842470 Atopic dermatitis ibudilast (multiple sclerosis/ amyotrophic lateral sclerosis), MediciNova Kyorin Pharmaceutical Co Ltd [00067] Ketas; MN-166; ibudilast; ibudilast (multiple sclerosis), MediciNova; ibudilast (multiple sclerosis/ amyotrophic lateral sclerosis), Neurological disease MediciNova PDE 4 inhibitors (asthma), Dainippon Sumitomo Dainippon Pharmaceutical Co Ltd [00068] OS-0217; PDE 4 inhibitors (asthma), Dainippon; PDE 4 inhibitors (asthma), Dainippon Sumitomo Asthma oglemilast Glenmark Pharmaceuticals Ltd [00069] GRC-3886; oglemilast; oglemilast (oral, COPD/asthma), Glenmark Asthma; Chronic obstructive pulmonary disease; Rheumatoid arthritis R-1627 Roche R-1627 Alzheimers Holding AG disease ND-1510 Neuro3d SA ND-1510 Depression ND-1251 Neuro3d SA ND-1251 Depression PDE4 inhibitors Purdue PDE4 inhibitors Asthma (asthma), Purdue Pharma LP (asthma), Purdue WAY-122331 Wyeth- Ayerst Pharmaceuticals Inc [00070] WAY-122331 Cardiac failure GRC-3566 Glenmark GRC-3566 Asthma; Pharmaceuticals Chronic Ltd obstructive pulmonary disease tofimilast Pfizer Inc [00071] CP-325366; tofimilast Allergy; Respiratory disease BAY-61-9987 Bayer AG BAY-61-9987; Chronic low affinity obstructive phosphodiesterase pulmonary 4 inhibitor, Bayer disease; Respiratory disease rolipram Bayer Schering Pharma AG [00072] ME-3167; ZK-62711; rolipram Asthma; Depression; HIV infection; Multiple sclerosis; Neurodegenerative disease; Tardive dyskinesia MEM-1414 Memory MEM-1414; Alzheimers Pharmaceuticals PDE 4 inhibitor disease; Corp (Alzheimer's), Asthma Memory; PDE 4 inhibitor (Alzheimer's), Memory/Roche; R-1533 adenosine A3 antagonists, Novartis Novartis AG [00073] CGH-2466; CGS-2466; adenosine A3 antagonists, Novartis Asthma RPL-554 King's College London [00074] PDE 3/PDE 4 inhibitors, Kings College; PDE3/4 inhibitors (nasal, respiratory disease), Verona Pharma; PDE3/4 inhibitors (respiratory therapeutics), Rhinopharma; RPL-554; RPL-565; VMX-554; VMX-565; VRP-554; dual MRP4 Allergic rhinitis and PDE3/4 inhibitors (nasal, respiratory disease), Verona Pharma; trequinsin analogs (respiratory therapeutics), Kings College/Vernalis/ Rhinopharma CT-5357 UCB Celltech [00075] CT-5357 Inflammatory disease etazolate Diaxonhit [00076] EHT-0202; SQ-20009; etazolate; etazolate hydrochloride Alzheimers disease; Motor neurone disease; Neurodegenerative disease Org-30029 MSD OSS BV [00077] Org-30029 Asthma; Cardiac failure PDE4 inhibitors (respiratory tract inflammation), Zambon Zambon Co SpA [00078] PDE4 inhibitors (respiratory tract inflammation), Zambon; Z-15370; Z-15370A Respiratory tract inflammation Orp-20241 MSD OSS BV [00079] Org-20241 Asthma PDE3/PDE4 inhibitors (inflammatory diseases), Leiden/ Amsterdam Center for Drug Research/Altana Leiden/ Amsterdam Center for Drug Research [00080] PDE3/PDE4 inhibitors (inflammatory diseases), Leiden/Amsterdam Center for Drug Research/Altana; PDE3/PDE4 inhibitors (inflammatory diseases), Leiden/ Amsterdam Center for Drug Research/ Byk Gulden Asthma; Rheumatoid arthritis arofylline Almirall Prodesfarma SA [00081] LAS-31025; arofylline Asthma; Bronchitis; Chronic obstructive airway disease KW-4490 Kyowa Hakko Kogyo Co Ltd [00082] KW-4490 Asthma HT-0712 Inflazyme Pharmaceuticals [00083] HT-0712; IPL-455903; small-molecule PDE4 inhibitors (memory disorders), Inflazyme/ Helicon Amnesia; Cognitive disorder PDE 4 inhibitors (asthma/COPD/ rheumatoid arthritis), Merck Frosst UCB Celltech [00084] CT-2450; CT-2820; CT-3883; CT-5210; L-454560; L-787258; L-791943; L-826141; L-869298; MK-0359; PDE 4 inhibitors (asthma/COPD/ rheumatoid arthritis), Merck Frosst; PDE 4 inhibitors, Celltech/Merck Frosst Asthma; Chronic obstructive pulmonary disease; Rheumatoid arthritis PDE inhibitors, VIVUS Inc PDE 3 inhibitors, Erectile Vivus Vivus dysfunction PDE 4 inhibitors, Vivus; PDE inhibitors, Vivus; PDE5 inhibitors, Vivus; erectile dysfunction therapy, Vivus OX-914 Inflazyme BLX-028914; Asthma; Pharmaceuticals BLX-914; IPL-4088; Chronic IPL-4182; IPL-42 obstructive series; IPL-4722; pulmonary OX-914; PDE4 disease; inhibitors Inflammatory (inflammation), disease; Biolipox; Seasonal PDE4 inhibitors allergic (inflammation), rhinitis Inflazyme; PDE4 inhibitors (inflammation), Orexo SDZ-PDI-747 Novartis AG [00085] SDZ-PDI-747 Atopic dermatitis AP-0679 The Green AP-0679 Asthma Cross Corp Sch-351591 UCB Celltech [00086] D-4396; PDE 4 inhibitors, Schering- Plough/Celltech; PDE 4 inhibitors, Schering- Plough/ Chiroscience; Sch-351591; Sch-365351 Asthma; Chronic obstructive pulmonary disease; Inflammatory disease TA-7906 Tanabe Seiyaku Co Ltd [00087] PDE4 inhibitor (skin disease), Maruho; PDE4 inhibitors (inflammation), Tanabe Seiyaku; T-2585; T-2585.HCl; TA-7906 Atopic dermatitis; Dermatological disease; Inflammatory disease PDE4/MMP inhibitors, Rhone- Poulenc Rhone- Poulenc Rorer Inc [00088] HMR-1571; PDE4/MMP inhibitors, Rhone- Poulenc Atherosclerosis; Atopic dermatitis; Multiple sclerosis; Psoriasis; Rheumatoid arthritis lirimilast Bayer AG [00089] BAY-19-8004; lirimilast Asthma; Chronic obstructive pulmonary disease daxalipram Bayer Schering Pharma AG [00090] Mesopram; PDE 4 inhibitor (multiple sclerosis), Schering AG; SH-636; ZK-117137; daxalipram Multiple sclerosis roflumilast Takeda GmbH [00091] APTA-2217; B9302-107; BY-217; BYK-20869; Daliresp; Dalveza; Daxas; Libertek; Xevex; roflumilast; roflumist Non-insulin dependent diabetes; Pulmonary fibrosis PDE 4 inhibitors (asthma), Novartis Novartis UK Ltd [00092] NVP-ABE-171; PDE 4 inhibitors (asthma), Novartis; rolipram analogs, Novartis Asthma; Chronic obstructive pulmonary disease PDE III/IV Novartis PDE III/IV Asthma; inhibitors Pharma AG inhibitors, Inflammatory Novartis Novartis disease SeICIDs Celcrene Celgene Corp [00093] CC-10036; CC-10083; CC-110007; CC-110036; CC-110037; CC-110038; CC-110049; CC-110052; CC-110083; CC-11069; CC-111050; CC-13039; CC-14046; CC-17034; CC-17035; Autoimmune disease; Cancer; Congestive heart failure; Inflammatory disease; Respiratory disease CC-17075; CC-17085; CC-18062; CC-7075; PDE4/TNFalpha inhibitors, Celgene; SeICIDs, Celgene; selective cytokine inhibitory drugs, Celgene RPR-117658 Rhone- Poulenc Rorer Ltd [00094] RPR-117658 Inflammatory disease AWD-12-281 (inhaled), elbion/ GlaxoSmithKline; ASTA Medica AG [00095] 842470; AWD-12-281; AWD-12-281 (COPD), elbion/ GlaxoSmithKline AWD-12-281 (asthma), elbion/ GlaxoSmithKline; AWD-12-281 (inhaled), elbion/ GlaxoSmithKline; Asthma; Chronic obstructive pulmonary disease AWD-12-343; GW-842470 256066 (asthma, COPD, inhalant formulation), GlaxoSmithKline SmithKline Beecham Pharmaceuticals [00096] 256066; 256066 (asthma, COPD, inhalant formulation), GlaxoSmithKline; GSK-256066; GSK-256066 (asthma, COPD, inhalant formulation), GlaxoSmithKline; PDE 4 inhibitors Asthma; Chronic obstructive pulmonary disease; Inflammatory disease (inhaled, COPD/asthma/ allergic rhinitis), GlaxoSmithKline; SB-207499 analogs, GSK PDE4 inhibitors, Aventis PDE4 inhibitors, Autoimmune Aventis Pharma AG Aventis; disease PDE4 inhibitors, RPR; PDE4 inhibitors, Rhone- Poulenc Rorer arofylline derivatives, Almirall Almirall Prodesfarma SA [00097] arofylline derivatives, Almirall Asthma; Inflammatory disease RPR-132294 Rhone- Poulenc Rorer Ltd [00098] RPR-132294; RPR-132703 Respiratory disease ibudilast eye drops (ocular allergy), MSD Japan/Kyorin Kyorin Pharmaceutical Co Ltd [00099] Eyevinal; KC-404; Ketas (ocular); ibudilast; ibudilast eye drops (ocular allergy), Banyu/Kyorin; ibudilast eye drops (ocular allergy), MSD Japan/ Kyorin PDE 4 inhibitors (2), Pfizer Pfizer Inc [00100] CI-1018; CI-1044; PD-168787; PD-189659; PD-190036; PD-190749; PDE 4 inhibitors (2), Pfizer Asthma; Inflammatory disease YM-976 Yamanouchi Pharmaceutical Co Ltd [00101] PDE IV inhibitors, Yamanouchi; YM-976; phosphodiesterase inhibitors, Yamanouchi Asthma XT-611 Kanazawa University [00102] PDE IV inhibitor, Kanazawa University; XT-611 Osteoporosis losartan Almirall losartan Asthma derivatives, Prodesfarma derivatives, Almirall SA Almirall DWP-205 derivatives, Daewoong Daewoong Pharmaceutical Co Ltd [00103] DWP-205 derivatives, Daewoong; DWP-205297; phosphodiesterase 4 inhibitors, Daewoong Arthritis; Asthma WAY-126120 Wyeth-Ayerst PDE IV inhibitor, Asthma Pharmaceuticals Wyeth-Ayerst; Inc WAY-126120 YM-58997 Yamanouchi Pharmaceutical Co Ltd [00104] YM-58997 Asthma CP-293321 Pfizer Inc CP-293321 Inflammatory disease V-11294A Napp Pharmaceutical Group Ltd [00105] V-11294A; rolipram derivatives, Napp Depression; Inflammatory disease CH-3697 Chiroscience CH-3697 Asthma R&D Ltd CP-353164 Pfizer Inc [00106] CP-353164 Rheumatoid arthritis atizoram Pfizer Inc [00107] CP-80633; atizoram Asthma; Dermatitis D-4418 Chiroscience R&D Ltd [00108] D-4418 Asthma; Inflammatory disease RPR-114597 Rhone- Poulenc Rorer Inc [00109] RPR-114597 Inflammatory disease PDE 4 inhibitors (inflammation), Eli Lilly ICOS Corp [00110] IC-197; IC-246; IC-247; IC-485; IC-86518; IC-86518/ IC-86521; IC-86521; PDE 4 inhibitors (inflammation), Eli Lilly; PDE 4 inhibitors, ICOS Chronic obstructive pulmonary disease; Inflammatory disease; Rheumatoid arthritis PDE 4 inhibitors, Pfizer Pfizer Inc [00111] BHN; CP-220629; PDE 4 inhibitors, Pfizer; UK-500001 Asthma; Chronic obstructive pulmonary disease ZL-n-91 Guanqzhou ZL-n-91, Guangzhou Sinogen Guangzhou Sinogen Pharmaceutical Sinogen Pharmaceutical Co Ltd Pharmaceutical D-22888 ASTA Medica AG [00112] AWD-12-232; D-22888 Allergy; Asthma PDE4 PDE4 Takeda PDE4 inhibitor inhibitor Pharmaceutical (diabetic (diabetic Co Ltd nephropathy), nephropathy), Takeda Takeda Pharmaceutical Pharmaceutical GW-3600 GlaxoSmithKline Inc [00113] GW-3600; phosphodiesterase 4 inhibitor, Glaxo Asthma; Inflammatory disease; Rheumatoid arthritis [00114]

[0945] Additional examples of a small molecule that is a PDE4 inhibitor include: Apremilast (CC-10004; CC-110004; CDC-104; Otezla; lead seCID (2); seCID); CC-1088 (CC-1088; CC-5048; CC-801; CDC-801; lead SelCID (1)); Tetomilast (OPC-6535); KF-19514; PF-06266047; SKF-107806; PDB-093; Tolafentrine (BY-4070); TAK-648; CH-928; CH-673; CH-422; ABI-4 (18F-PF-06445974; Fluorine-18-PF-06445974); roflumilast; Roflumilast N-oxide (APTA-2217; B9302-107; BY-217; BYK-20860; Daliresp; Dalveza; Daxas; Libertek; Xevex; roflumist); NVP-ABE-171; BYK-321084; WAY-127093B; NCS-613; SDZ-ISQ-844; GS-5759; Ro-20-1724; Hemay-005; KCA-1490; TVX-2706; Nitraquazone; Filaminast (PDA-641; WAY-PDA-641); LASSBio-596; ASP-3258; TAS-203; AN-2889; AN-5322; AN-6414; AN-6415; Iotamilast (E-6005; RVT-501); GPD-1116; Cipamfylline (BRL-61063; HEP-688); MNP-001; MS-23; MSP-001; K-34; KF-66490; AL-38583 (cilomast); ZL-N-91; Almirall; CDP-840; GSK-356728; Cilomilast (Ariflo; SB-207499); OCID-2987; AN-2898; CBS-3595; ASP-9831 (ASP9831); E-4021 (4-Piperidinecarboxylic acid, 1-[4-[(1,3-benzodioxol-5-ylmethyl)amino]-6-chloro-2-quinazolinyl]); Piclamilast (RP-73401; RPR-73401); CD-160130; GSK-256066 (256066); 4AZA-PDE4; YM-393059; Revamilast (GRC-4039); AN-2728 (PF-06930164; crisaborole (Eucrisa)); MK-0952 (MK-952); Ibudilast (AV-411; MN-166; KC-404); GP-0203; ELB-526; Theophylline (Teonova); CHF-6001 (CHF-5480); Elbimilast (AWD-12-353; ELB-353; ronomilast); AWD-12-281 (842470); OS-0217; Oglemilast (GRC-3886); R-1627; ND-1510; ND-1251; WAY-122331; GRC-3566; Tofimilast (CP-325366); BAY-61-9987; Rolipram (ME-3167; ZK-62711); MEM-1414 (R-1533); Adenosine A3 antagonists (CGH-2466); RPL-554 (RPL-565; VMX-554; VMX-565; VRP-554; trequinsin analog); CT-5357; Etazolate (EHT-0202; SQ-20009; etazolate hydrochloride); Z-15370 (Z-15370A); Org-30029; Org-20241; Arofylline (LAS-31025); Arofylline derivatives; KW-4490; HT-0712 (IPL-455903); HT-0712; IPL-455903; CT-2450; CT-2820; CT-3883; CT-5210; L-454560; L-787258; L-791943; L-826141; L-869298; MK-0359; OX-914 (BLX-028914; BLX-914; IPL-4088; IPL-4182; IPL-4722); SDZ-PDI-747; AP-0679; Sch-351591 (D-4396; Sch-365351); TA-7906 (T-2585; TA-7906); HMR-1571; Lirimilast (BAY-19-8004); Daxalipram (Mesopram; SH-636; ZK-117137); SelCIs (CC-10036; CC-10083; CC-110007; CC-110036; CC-110037; CC-110038; CC-110049; CC-110052; CC-110083; CC-11069; CC-111050; CC-13039; CC-14046; CC-17034; CC-17035; CC-17075; CC-17085; CC-18062; CC-7075); RPR-117658; AWD-12-281 (842470; AWD-12-343; GW842470X); 256066 (GSK-256066; SB-207499); RPR-132294 (RPR-132703); CI-1018; CI-1044; PD-168787; PD-189659; PD-190036; PD-190749; YM-976; XT-611; Losartan derivatives; DWP-205 derivatives (DWP-205297); WAY-126120; YM-58997; CP-293321; V-11294A; CH-3697; CP-353164; Atizoram (CP-80633); D-4418; RPR-114597; IC-197; IC-246; IC-247; IC-485; IC-86518; IC-86518/IC-86521; IC-86521; CP-220629; ZL-n-91; D-22888 (AWD-12-232); GW-3600; GSK356278; TPI 1100; BPN14770; and MK-0873. See, e.g., Schafter et al. (2014) Cellular Signaling 26(9): 2016-2029); Gurney et al. (2011) Handb Exp Pharmacol 204: 167-192; Spadaccini et al. (2017) Intl J Mol Sciences 18: 1276; Bickston et al. (2012) Expert Opinion Invest Drugs 21:12, 1845-1849; Keshavarzian et al. (2007) Expert Opinion Invest Drugs 16:9, 1489-1506.

[0946] Additional examples of small molecules that are PDE4 inhibitors are described in, e.g., U.S. Patent Application Publication Nos. 2017/0348311, 20176/0319558, 2016/0213642, 2015/0328187, 2015/0306079, 2015/0272949, 2015/0272936, 2015/0080359, 2015/0051254, 2014/0350035, 2014/0148420, 2014/0121221, 2013/0252928, 2013/0237527, 2013/0225609, 2012/0309726, 2012/0196867, 2012/0088743, 2012/0059031, 2012/0035143, 2012/0028932, 2011/0021478, 2011/0021476, 2010/0234382, 2010/0129363, 2010/0069392, 2010/0056604, 2010/0048616, 2010/0048615, 2009/0099148, 2009/0093503, 2008/0287522, 2008/0255209, 2008/0255186, 2008/0221111, 2007/0232637, 2007/0208181, 2007/0167489, 2006/0269600, 2006/0183764, 2006/0154934, 2006/0094723, 2006/0079540, 2005/0267135, 2005/0234238, 2005/0033521, 2003/0229134, 2003/0220352, 2003/0212112, 2003/0158189, 2003/0069260, 2003/0050329, 2002/0058687, and 2002/0028842. Additional examples of small molecules that are PDE4 inhibitors are known in the art.

Inhibitory Nucleic Acids

[0947] In some embodiments, a PDE4 inhibitor can be an inhibitory nucleic acid. In some embodiments, the inhibitory nucleic acid can be an antisense nucleic acid, a ribozyme, and a small interfering RNA (siRNA). Examples of aspects of these different oligonucleotides are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of PDE4 mRNA in a mammalian cell can be synthesized in vitro.

[0948] Inhibitory nucleic acids that can decrease the expression of PDE4 mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an PDE4 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 1-5).

TABLE-US-00025 HumanPDE4mRNATranscriptVariant1 (SEQIDNO:1) 1cggccgggcgcacccgcggggccctgggctcgctggcttgcgcgcagctgagcggggtgt 61aggttggaagggccagggccccctggggcgcaagtgggggccggcgccatggaacccccg 121accgtcccctcggaaaggagcctgtctctgtcactgcccgggccccgggagggccaggcc 181accctgaagcctcccccgcagcacctgtggcggcagcctcggacccccatccgtatccag 241cagcgcggctactccgacagcgcggagcgcgccgagcgggagcggcagccgcaccggccc 301atagagcgcgccgatgccatggacaccagcgaccggcccggcctgcgcacgacccgcatg 361tcctggccctcgtccttccatggcactggcaccggcagcggcggcgcgggcggaggcagc 421agcaggcgcttcgaggcagagaatgggccgacaccatctcctggccgcagccccctggac 481tcgcaggcgagcccaggactcgtgctgcacgccggggcggccaccagccagcgccgggag 541tccttcctgtaccgctcagacagcgactatgacatgtcacccaagaccatgtcccggaac 601tcatcggtcaccagcgaggcgcacgctgaagacctcatcgtaacaccatttgctcaggtg 661ctggccagcctccggagcgtccgtagcaacttctcactcctgaccaatgtgcccgttccc 721agtaacaagcggtccccgctgggcggccccacccctgtctgcaaggccacgctgtcagaa 781gaaacgtgtcagcagttggcccgggagactctggaggagctggactggtgtctggagcag 841ctggagaccatgcagacctatcgctctgtcagcgagatggcctcgcacaagttcaaaagg 901atgttgaaccgtgagctcacacacctgtcagaaatgagcaggtccggaaaccaggtctca 961gagtacatttccacaacattcctggacaaacagaatgaagtggagatcccatcacccacg 1021atgaaggaacgagaaaaacagcaagcgccgcgaccaagaccctcccagccgcccccgccc 1081cctgtaccacacttacagcccatgtcccaaatcacagggttgaaaaagttgatgcatagt 1141aacagcctgaacaactctaacattccccgatttggggtgaagaccgatcaagaagagctc 1201ctggcccaagaactggagaacctgaacaagtggggcctgaacatcttttgcgtgtcggat 1261tacgctggaggccgctcactcacctgcatcatgtacatgatattccaggagcgggacctg 1321ctgaagaaattccgcatccctgtggacacgatggtgacatacatgctgacgctggaggat 1381cactaccacgctgacgtggcctaccataacagcctgcacgcagctgacgtgctgcagtcc 1441acccacgtactgctggccacgcctgcactagatgcagtgttcacggacctggagattctc 1501gccgccctcttcgcggctgccatccacgatgtggatcaccctggggtctccaaccagttc 1561ctcatcaacaccaattcggagctggcgctcatgtacaacgatgagtcggtgctcgagaat 1621caccacctggccgtgggcttcaagctgctgcaggaggacaactgcgacatcttccagaac 1681ctcagcaagcgccagcggcagagcctacgcaagatggtcatcgacatggtgctggccacg 1741gacatgtccaagcacatgaccctcctggctgacctgaagaccatggtggagaccaagaaa 1801gtgaccagctcaggggtcctcctgctagataactactccgaccgcatccaggtcctccgg 1861aacatggtgcactgtgccgacctcagcaaccccaccaagccgctggagctgtaccgccag 1921tggacagaccgcatcatggccgagttcttccagcagggtgaccgagagcgcgagcgtggc 1981atggaaatcagccccatgtgtgacaagcacactgcctccgtggagaagtctcaggtgggt 2041tttattgactacattgtgcacccattgtgggagacctgggcggaccttgtccacccagat 2101gcccaggagatcttggacactttggaggacaaccgggactggtactacagcgccatccgg 2161cagagcccatctccgccacccgaggaggagtcaagggggccaggccacccacccctgcct 2221gacaagttccagtttgagctgacgctggaggaggaagaggaggaagaaatatcaatggcc 2281cagataccgtgcacagcccaagaggcattgactgcgcagggattgtcaggagtcgaggaa 2341gctctggatgcaaccatagcctgggaggcatccccggcccaggagtcgttggaagttatg 2401gcacaggaagcatccctggaggccgagctggaggcagtgtatttgacacagcaggcacag 2461tccacaggcagtgcacctgtggctccggatgagttctcgtcccgggaggaattcgtggtt 2521gctgtaagccacagcagcccctctgccctggctcttcaaagcccccttctccctgcttgg 2581aggaccctgtctgtttcagagcatgccccgggcctcccgggcctcccctccacggcggcc 2641gaggtggaggcccaacgagagcaccaggctgccaagagggcttgcagtgcctgcgcaggg 2701acatttggggaggacacatccgcactcccagctcctggtggcggggggtcaggtggagac 2761cctacctgatccccagacctctgtccctgttcccctccactcctcccctcactcccctgc 2821tcccccgaccacctcctcctctgcctcaaagactcttgtcctcttgtccctcctgagaaa 2881aaagaaaacgaaaagtggggtttttttctgttttctttttttcccctttccccctgcccc 2941cacccacggggcctttttttggaggtgggggctggggaatgaggggctgaggtcccggaa 3001gggattttatttttttgaattttaattgtaacatttttagaaaaagaacaaaaaaagaaa 3061aaaaaaagaaagaaacacagcaactgtagatgctcctgttcctggttcccgctttccact 3121tccaaatccctcccctcaccttcccccactgccccccaagttccaggctcagtcttccag 3181ccgcctggggagtctctacctgggcccaagcaggtgtggggcctccttctgggcttttct 3241tctgaatttagaggatttctagaacgtggtcaggaatagccattctaggcggggctgggg 3301ccagggtggggggcagtcactgtgggaggtcccagctccagcccccctctggtttgctgc 3361ctcctctcccctctaaaaaagtcttccgcttgattttgcacaatcccggcgatactcctg 3421gcgatactgactagaaagtcagggagctgggggagctgttcactttaggatacgggggtg 3481gtatggaagggagcgttcacaccgccagcctcgggcctgggatttgaggagggccctaga 3541cctcctccactctccatcccctttcccttccactttgggttcactttgaattttctccgt 3601tttttggggcagtggctctgatccactcacccccccgccccccgccccacttctagctgc 3661ttctcctcttgtttctgccttaataattcccacggccacaggcaagggggttgcagtggc 3721cgcctgcaccttggatgaggcagggccaggcgcccagaacccccatcctggccgcacccc 3781cctttccagggtcctccggaccccaccttccacactctgatcacagcccccctacctttt 3841gccctaggaggaagcaataatggtgtataccctcattctcattcctgggcagcccttcct 3901tccaccctggcaccaaaataatttctcctccatccgtaccttgcctagcctctccctctc 3961ccccagctagtccctgagcaatacggcagacagatgcaagaccatttttctccaagccat 4021gggggactgtttggaaggaaagccccctctctccctcctcccctcgccctcggcctggtt 4081ctgcagctggaccgacctcattcatcgcctgccccctacccaattctgagcacacggtac 4141tgtagcccccagttcctccctagccttccatccctctgtccaccccagggggaggtaacc 4201ccgcactcacactcccttgatgctgtctgtacagggttcatattttgtagcgaaagtcgt 4261ttttgtcccagccggcgatcggagtgggccttttctttctttttgttcattctttacctt 4321tttttcttttctttctttcttttttgtacatactgtaaggttggtttgtaaattattcta 4381cggaggcaaaaagggaaaataaaaacttgcccttccctggctgacccagtcgggaaggta 4441gggaaggaggtctcccgttgggagagtctctgttcctgctgtattatacaaactgtacca 4501tagtcctgggaaaagggtggactcaccgctgttgttttatgggaagtcgtgtcatcctag 4561gggttggggctgggcagagcctgtcccctccccccttctccaggagccagggggtgactg 4621gagagacagacccacccccaagcagggctcctctccccagggtgagcacaggacctctgt 4681aagctgcttgtgtattgtccactttgacgatcagtcattcggtccgttgatcaataatcc 4741ttcgatcttgtctccaattaaaccgaggctttcaccgataaaaaaaaaaaaaaa HumanPDE4mRNATranscriptVariant2 (SEQIDNO:2) 1atggcgcggccgcgcggcctaggccgcatcccggagctgcaactggtggccttcccggtg 61gcggtggcggctgaggacgaggcgttcctgcccgagcccctggccccgcgcgcgccccgc 121cgcccgcgttcgccgccctcctcgcccgtcttcttcgccagcccgtccccaactttccgc 181agacgccttcggcttctccgcagctgccaggatttgggccgccaggcttgggctggggct 241ggcttcgaggcagagaatgggccgacaccatctcctggccgcagccccctggactcgcag 301gcgagcccaggactcgtgctgcacgccggggcggccaccagccagcgccgggagtccttc 361ctgtaccgctcagacagcgactatgacatgtcacccaagaccatgtcccggaactcatcg 421gtcaccagcgaggcgcacgctgaagacctcatcgtaacaccatttgctcaggtgctggcc 481agcctccggagcgtccgtagcaacttctcactcctgaccaatgtgcccgttcccagtaac 541aagcggtccccgctgggcggccccacccctgtctgcaaggccacgctgtcagaagaaacg 601tgtcagcagttggcccgggagactctggaggagctggactggtgtctggagcagctggag 661accatgcagacctatcgctctgtcagcgagatggcctcgcacaagttcaaaaggatgttg 721aaccgtgagctcacacacctgtcagaaatgagcaggtccggaaaccaggtctcagagtac 781atttccacaacattcctggacaaacagaatgaagtggagatcccatcacccacgatgaag 841gaacgagaaaaacagcaagcgccgcgaccaagaccctcccagccgcccccgccccctgta 901ccacacttacagcccatgtcccaaatcacagggttgaaaaagttgatgcatagtaacagc 961ctgaacaactctaacattccccgatttggggtgaagaccgatcaagaagagctcctggcc 1021caagaactggagaacctgaacaagtggggcctgaacatcttttgcgtgtcggattacgct 1081ggaggccgctcactcacctgcatcatgtacatgatattccaggagcgggacctgctgaag 1141aaattccgcatccctgtggacacgatggtgacatacatgctgacgctggaggatcactac 1201cacgctgacgtggcctaccataacagcctgcacgcagctgacgtgctgcagtccacccac 1261gtactgctggccacgcctgcactagatgcagtgttcacggacctggagattctcgccgcc 1321ctcttcgcggctgccatccacgatgtggatcaccctggggtctccaaccagttcctcatc 1381aacaccaattcggagctggcgctcatgtacaacgatgagtcggtgctcgagaatcaccac 1441ctggccgtgggcttcaagctgctgcaggaggacaactgcgacatcttccagaacctcagc 1501aagcgccagcggcagagcctacgcaagatggtcatcgacatggtgctggccacggacatg 1561tccaagcacatgaccctcctggctgacctgaagaccatggtggagaccaagaaagtgacc 1621agctcaggggtcctcctgctagataactactccgaccgcatccaggtcctccggaacatg 1681gtgcactgtgccgacctcagcaaccccaccaagccgctggagctgtaccgccagtggaca 1741gaccgcatcatggccgagttcttccagcagggtgaccgagagcgcgagcgtggcatggaa 1801atcagccccatgtgtgacaagcacactgcctccgtggagaagtctcaggtgggttttatt 1861gactacattgtgcacccattgtgggagacctgggcggaccttgtccacccagatgcccag 1921gagatcttggacactttggaggacaaccgggactggtactacagcgccatccggcagagc 1981ccatctccgccacccgaggaggagtcaagggggccaggccacccacccctgcctgacaag 2041ttccagtttgagctgacgctggaggaggaagaggaggaagaaatatcaatggcccagata 2101ccgtgcacagcccaagaggcattgactgcgcagggattgtcaggagtcgaggaagctctg 2161gatgcaaccatagcctgggaggcatccccggcccaggagtcgttggaagttatggcacag 2221gaagcatccctggaggccgagctggaggcagtgtatttgacacagcaggcacagtccaca 2281ggcagtgcacctgtggctccggatgagttctcgtcccgggaggaattcgtggttgctgta 2341agccacagcagcccctctgccctggctcttcaaagcccccttctccctgcttggaggacc 2401ctgtctgtttcagagcatgccccgggcctcccgggcctcccctccacggcggccgaggtg 2461gaggcccaacgagagcaccaggctgccaagagggcttgcagtgcctgcgcagggacattt 2521ggggaggacacatccgcactcccagctcctggtggcggggggtcaggtggagaccctacc 2581tgatccccagacctctgtccctgttcccctccactcctcccctcactcccctgctccccc 2641gaccacctcctcctctgcctcaaagactcttgtcctcttgtccctcctgagaaaaaagaa 2701aacgaaaagtggggtttttttctgttttctttttttcccctttccccctgcccccaccca 2761cggggcctttttttggaggtgggggctggggaatgaggggctgaggtcccggaagggatt 2821ttatttttttgaattttaattgtaacatttttagaaaaagaacaaaaaaagaaaaaaaaa 2881agaaagaaacacagcaactgtagatgctcctgttcctggttcccgctttccacttccaaa 2941tccctcccctcaccttcccccactgccccccaagttccaggctcagtcttccagccgcct 3001ggggagtctctacctgggcccaagcaggtgtggggcctccttctgggcttttcttctgaa 3061tttagaggatttctagaacgtggtcaggaatagccattctaggcggggctggggccaggg 3121tggggggcagtcactgtgggaggtcccagctccagcccccctctggtttgctgcctcctc 3181tcccctctaaaaaagtcttccgcttgattttgcacaatcccggcgatactcctggcgata 3241ctgactagaaagtcagggagctgggggagctgttcactttaggatacgggggtggtatgg 3301aagggagcgttcacaccgccagcctcgggcctgggatttgaggagggccctagacctcct 3361ccactctccatcccctttcccttccactttgggttcactttgaattttctccgttttttg 3421gggcagtggctctgatccactcacccccccgccccccgccccacttctagctgcttctcc 3481tcttgtttctgccttaataattcccacggccacaggcaagggggttgcagtggccgcctg 3541caccttggatgaggcagggccaggcgcccagaacccccatcctggccgcacccccctttc 3601cagggtcctccggaccccaccttccacactctgatcacagcccccctaccttttgcccta 3661ggaggaagcaataatggtgtataccctcattctcattcctgggcagcccttccttccacc 3721ctggcaccaaaataatttctcctccatccgtaccttgcctagcctctccctctcccccag 3781ctagtccctgagcaatacggcagacagatgcaagaccatttttctccaagccatggggga 3841ctgtttggaaggaaagccccctctctccctcctcccctcgccctcggcctggttctgcag 3901ctggaccgacctcattcatcgcctgccccctacccaattctgagcacacggtactgtagc 3961ccccagttcctccctagccttccatccctctgtccaccccagggggaggtaaccccgcac 4021tcacactcccttgatgctgtctgtacagggttcatattttgtagcgaaagtcgtttttgt 4081cccagccggcgatcggagtgggccttttctttctttttgttcattctttacctttttttc 4141ttttctttctttcttttttgtacatactgtaaggttggtttgtaaattattctacggagg 4201caaaaagggaaaataaaaacttgcccttccctggctgacccagtcgggaaggtagggaag 4261gaggtctcccgttgggagagtctctgttcctgctgtattatacaaactgtaccatagtcc 4321tgggaaaagggtggactcaccgctgttgttttatgggaagtcgtgtcatcctaggggttg 4381gggctgggcagagcctgtcccctccccccttctccaggagccagggggtgactggagaga 4441cagacccacccccaagcagggctcctctccccagggtgagcacaggacctctgtaagctg 4501cttgtgtattgtccactttgacgatcagtcattcggtccgttgatcaataatccttcgat 4561cttgtctccaattaaaccgaggctttcaccgataaaaaaaaaaaaaaa HumanPDE4mRNATranscriptVariant3 (SEQIDNO:3) 1atgcgctccggtgcagcgccccgggcccggccccggccccctgccctggcactgcccccc 61acgggccccgagtccctgacccacttccccttcagcgatgaggacacccgtcggcaccct 121ccgggcagatctgtcagcttcgaggcagagaatgggccgacaccatctcctggccgcagc 181cccctggactcgcaggcgagcccaggactcgtgctgcacgccggggcggccaccagccag 241cgccgggagtccttcctgtaccgctcagacagcgactatgacatgtcacccaagaccatg 301tcccggaactcatcggtcaccagcgaggcgcacgctgaagacctcatcgtaacaccattt 361gctcaggtgctggccagcctccggagcgtccgtagcaacttctcactcctgaccaatgtg 421cccgttcccagtaacaagcggtccccgctgggcggccccacccctgtctgcaaggccacg 481ctgtcagaagaaacgtgtcagcagttggcccgggagactctggaggagctggactggtgt 541ctggagcagctggagaccatgcagacctatcgctctgtcagcgagatggcctcgcacaag 601ttcaaaaggatgttgaaccgtgagctcacacacctgtcagaaatgagcaggtccggaaac 661caggtctcagagtacatttccacaacattcctggacaaacagaatgaagtggagatccca 721tcacccacgatgaaggaacgagaaaaacagcaagcgccgcgaccaagaccctcccagccg 781cccccgccccctgtaccacacttacagcccatgtcccaaatcacagggttgaaaaagttg 841atgcatagtaacagcctgaacaactctaacattccccgatttggggtgaagaccgatcaa 901gaagagctcctggcccaagaactggagaacctgaacaagtggggcctgaacatcttttgc 961gtgtcggattacgctggaggccgctcactcacctgcatcatgtacatgatattccaggag 1021cgggacctgctgaagaaattccgcatccctgtggacacgatggtgacatacatgctgacg 1081ctggaggatcactaccacgctgacgtggcctaccataacagcctgcacgcagctgacgtg 1141ctgcagtccacccacgtactgctggccacgcctgcactagatgcagtgttcacggacctg 1201gagattctcgccgccctcttcgcggctgccatccacgatgtggatcaccctggggtctcc 1261aaccagttcctcatcaacaccaattcggagctggcgctcatgtacaacgatgagtcggtg 1321ctcgagaatcaccacctggccgtgggcttcaagctgctgcaggaggacaactgcgacatc 1381ttccagaacctcagcaagcgccagcggcagagcctacgcaagatggtcatcgacatggtg 1441ctggccacggacatgtccaagcacatgaccctcctggctgacctgaagaccatggtggag 1501accaagaaagtgaccagctcaggggtcctcctgctagataactactccgaccgcatccag 1561gtcctccggaacatggtgcactgtgccgacctcagcaaccccaccaagccgctggagctg 1621taccgccagtggacagaccgcatcatggccgagttcttccagcagggtgaccgagagcgc 1681gagcgtggcatggaaatcagccccatgtgtgacaagcacactgcctccgtggagaagtct 1741caggtgggttttattgactacattgtgcacccattgtgggagacctgggcggaccttgtc 1801cacccagatgcccaggagatcttggacactttggaggacaaccgggactggtactacagc 1861gccatccggcagagcccatctccgccacccgaggaggagtcaagggggccaggccaccca 1921cccctgcctgacaagttccagtttgagctgacgctggaggaggaagaggaggaagaaata 1981tcaatggcccagataccgtgcacagcccaagaggcattgactgcgcagggattgtcagga 2041gtcgaggaagctctggatgcaaccatagcctgggaggcatccccggcccaggagtcgttg 2101gaagttatggcacaggaagcatccctggaggccgagctggaggcagtgtatttgacacag 2161caggcacagtccacaggcagtgcacctgtggctccggatgagttctcgtcccgggaggaa 2221ttcgtggttgctgtaagccacagcagcccctctgccctggctcttcaaagcccccttctc 2281cctgcttggaggaccctgtctgtttcagagcatgccccgggcctcccgggcctcccctcc 2341acggcggccgaggtggaggcccaacgagagcaccaggctgccaagagggcttgcagtgcc 2401tgcgcagggacatttggggaggacacatccgcactcccagctcctggtggcggggggtca 2461ggtggagaccctacctgatccccagacctctgtccctgttcccctccactcctcccctca 2521ctcccctgctcccccgaccacctcctcctctgcctcaaagactcttgtcctcttgtccct 2581cctgagaaaaaagaaaacgaaaagtggggtttttttctgttttctttttttcccctttcc 2641ccctgcccccacccacggggcctttttttggaggtgggggctggggaatgaggggctgag 2701gtcccggaagggattttatttttttgaattttaattgtaacatttttagaaaaagaacaa 2761aaaaagaaaaaaaaaagaaagaaacacagcaactgtagatgctcctgttcctggttcccg 2821ctttccacttccaaatccctcccctcaccttcccccactgccccccaagttccaggctca 2881gtcttccagccgcctggggagtctctacctgggcccaagcaggtgtggggcctccttctg 2941ggcttttcttctgaatttagaggatttctagaacgtggtcaggaatagccattctaggcg 3001gggctggggccagggtggggggcagtcactgtgggaggtcccagctccagcccccctctg 3061gtttgctgcctcctctcccctctaaaaaagtcttccgcttgattttgcacaatcccggcg 3121atactcctggcgatactgactagaaagtcagggagctgggggagctgttcactttaggat 3181acgggggtggtatggaagggagcgttcacaccgccagcctcgggcctgggatttgaggag 3241ggccctagacctcctccactctccatcccctttcccttccactttgggttcactttgaat 3301tttctccgttttttggggcagtggctctgatccactcacccccccgccccccgccccact 3361tctagctgcttctcctcttgtttctgccttaataattcccacggccacaggcaagggggt 3421tgcagtggccgcctgcaccttggatgaggcagggccaggcgcccagaacccccatcctgg 3481ccgcacccccctttccagggtcctccggaccccaccttccacactctgatcacagccccc 3541ctaccttttgccctaggaggaagcaataatggtgtataccctcattctcattcctgggca 3601gcccttccttccaccctggcaccaaaataatttctcctccatccgtaccttgcctagcct 3661ctccctctcccccagctagtccctgagcaatacggcagacagatgcaagaccatttttct 3721ccaagccatgggggactgtttggaaggaaagccccctctctccctcctcccctcgccctc 3781ggcctggttctgcagctggaccgacctcattcatcgcctgccccctacccaattctgagc 3841acacggtactgtagcccccagttcctccctagccttccatccctctgtccaccccagggg 3901gaggtaaccccgcactcacactcccttgatgctgtctgtacagggttcatattttgtagc 3961gaaagtcgtttttgtcccagccggcgatcggagtgggccttttctttctttttgttcatt 4021ctttacctttttttcttttctttctttcttttttgtacatactgtaaggttggtttgtaa 4081attattctacggaggcaaaaagggaaaataaaaacttgcccttccctggctgacccagtc 4141gggaaggtagggaaggaggtctcccgttgggagagtctctgttcctgctgtattatacaa 4201actgtaccatagtcctgggaaaagggtggactcaccgctgttgttttatgggaagtcgtg 4261tcatcctaggggttggggctgggcagagcctgtcccctccccccttctccaggagccagg 4321gggtgactggagagacagacccacccccaagcagggctcctctccccagggtgagcacag 4381gacctctgtaagctgcttgtgtattgtccactttgacgatcagtcattcggtccgttgat 4441caataatccttcgatcttgtctccaattaaaccgaggctttcaccgataaaaaaaaaaaa 4501aaa HumanPDE4mRNATranscriptVariant4 (SEQIDNO:4) 1tccgcagcctcctcctgggacccttgccctgcccccctcccatgggcacggaccccccac 61cgcctccacccactgccgcgggggggcccgttggggcccagggctggcgggccatgtaac 121cagggctgctgctgggagcgcggaggggaagggagcccccagccctgctgggccggccca 181ggcccctccgcggctcccccttccactacccacctgcccggcaccccctccccagtggtt 241gttaaccccgggactccccaagcccagcctctgtgtgcagcagccccaggcgggctaagt 301ctccaagatgcccttggtggatttcttctgcgagacctgctctaagccttggctggtggg 361ctggtgggaccagttcaaaaggatgttgaaccgtgagctcacacacctgtcagaaatgag 421caggtccggaaaccaggtctcagagtacatttccacaacattcctggacaaacagaatga 481agtggagatcccatcacccacgatgaaggaacgagaaaaacagcaagcgccgcgaccaag 541accctcccagccgcccccgccccctgtaccacacttacagcccatgtcccaaatcacagg 601gttgaaaaagttgatgcatagtaacagcctgaacaactctaacattccccgatttggggt 661gaagaccgatcaagaagagctcctggcccaagaactggagaacctgaacaagtggggcct 721gaacatcttttgcgtgtcggattacgctggaggccgctcactcacctgcatcatgtacat 781gatattccaggagcgggacctgctgaagaaattccgcatccctgtggacacgatggtgac 841atacatgctgacgctggaggatcactaccacgctgacgtggcctaccataacagcctgca 901cgcagctgacgtgctgcagtccacccacgtactgctggccacgcctgcactagatgcagt 961gttcacggacctggagattctcgccgccctcttcgcggctgccatccacgatgtggatca 1021ccctggggtctccaaccagttcctcatcaacaccaattcggagctggcgctcatgtacaa 1081cgatgagtcggtgctcgagaatcaccacctggccgtgggcttcaagctgctgcaggagga 1141caactgcgacatcttccagaacctcagcaagcgccagcggcagagcctacgcaagatggt 1201catcgacatggtgctggccacggacatgtccaagcacatgaccctcctggctgacctgaa 1261gaccatggtggagaccaagaaagtgaccagctcaggggtcctcctgctagataactactc 1321cgaccgcatccaggtcctccggaacatggtgcactgtgccgacctcagcaaccccaccaa 1381gccgctggagctgtaccgccagtggacagaccgcatcatggccgagttcttccagcaggg 1441tgaccgagagcgcgagcgtggcatggaaatcagccccatgtgtgacaagcacactgcctc 1501cgtggagaagtctcaggtgggttttattgactacattgtgcacccattgtgggagacctg 1561ggcggaccttgtccacccagatgcccaggagatcttggacactttggaggacaaccggga 1621ctggtactacagcgccatccggcagagcccatctccgccacccgaggaggagtcaagggg 1681gccaggccacccacccctgcctgacaagttccagtttgagctgacgctggaggaggaaga 1741ggaggaagaaatatcaatggcccagataccgtgcacagcccaagaggcattgactgcgca 1801gggattgtcaggagtcgaggaagctctggatgcaaccatagcctgggaggcatccccggc 1861ccaggagtcgttggaagttatggcacaggaagcatccctggaggccgagctggaggcagt 1921gtatttgacacagcaggcacagtccacaggcagtgcacctgtggctccggatgagttctc 1981gtcccgggaggaattcgtggttgctgtaagccacagcagcccctctgccctggctcttca 2041aagcccccttctccctgcttggaggaccctgtctgtttcagagcatgccccgggcctccc 2101gggcctcccctccacggcggccgaggtggaggcccaacgagagcaccaggctgccaagag 2161ggcttgcagtgcctgcgcagggacatttggggaggacacatccgcactcccagctcctgg 2221tggcggggggtcaggtggagaccctacctgatccccagacctctgtccctgttcccctcc 2281actcctcccctcactcccctgctcccccgaccacctcctcctctgcctcaaagactcttg 2341tcctcttgtccctcctgagaaaaaagaaaacgaaaagtggggtttttttctgttttcttt 2401ttttcccctttccccctgcccccacccacggggcctttttttggaggtgggggctgggga 2461atgaggggctgaggtcccggaagggattttatttttttgaattttaattgtaacattttt 2521agaaaaagaacaaaaaaagaaaaaaaaaagaaagaaacacagcaactgtagatgctcctg 2581ttcctggttcccgctttccacttccaaatccctcccctcaccttcccccactgcccccca 2641agttccaggctcagtcttccagccgcctggggagtctctacctgggcccaagcaggtgtg 2701gggcctccttctgggcttttcttctgaatttagaggatttctagaacgtggtcaggaata 2761gccattctaggcggggctggggccagggtggggggcagtcactgtgggaggtcccagctc 2821cagcccccctctggtttgctgcctcctctcccctctaaaaaagtcttccgcttgattttg 2881cacaatcccggcgatactcctggcgatactgactagaaagtcagggagctgggggagctg 2941ttcactttaggatacgggggtggtatggaagggagcgttcacaccgccagcctcgggcct 3001gggatttgaggagggccctagacctcctccactctccatcccctttcccttccactttgg 3061gttcactttgaattttctccgttttttggggcagtggctctgatccactcacccccccgc 3121cccccgccccacttctagctgcttctcctcttgtttctgccttaataattcccacggcca 3181caggcaagggggttgcagtggccgcctgcaccttggatgaggcagggccaggcgcccaga 3241acccccatcctggccgcacccccctttccagggtcctccggaccccaccttccacactct 3301gatcacagcccccctaccttttgccctaggaggaagcaataatggtgtataccctcattc 3361tcattcctgggcagcccttccttccaccctggcaccaaaataatttctcctccatccgta 3421ccttgcctagcctctccctctcccccagctagtccctgagcaatacggcagacagatgca 3481agaccatttttctccaagccatgggggactgtttggaaggaaagccccctctctccctcc 3541tcccctcgccctcggcctggttctgcagctggaccgacctcattcatcgcctgcccccta 3601cccaattctgagcacacggtactgtagcccccagttcctccctagccttccatccctctg 3661tccaccccagggggaggtaaccccgcactcacactcccttgatgctgtctgtacagggtt 3721catattttgtagcgaaagtcgtttttgtcccagccggcgatcggagtgggccttttcttt 3781ctttttgttcattctttacctttttttcttttctttctttcttttttgtacatactgtaa 3841ggttggtttgtaaattattctacggaggcaaaaagggaaaataaaaacttgcccttccct 3901ggctgacccagtcgggaaggtagggaaggaggtctcccgttgggagagtctctgttcctg 3961ctgtattatacaaactgtaccatagtcctgggaaaagggtggactcaccgctgttgtttt 4021atgggaagtcgtgtcatcctaggggttggggctgggcagagcctgtcccctccccccttc 4081tccaggagccagggggtgactggagagacagacccacccccaagcagggctcctctcccc 4141agggtgagcacaggacctctgtaagctgcttgtgtattgtccactttgacgatcagtcat 4201tcggtccgttgatcaataatccttcgatcttgtctccaattaaaccgaggctttcaccga 4261taaaaaaaaaaaaaaa HumanPDE4mRNATranscriptVariant5 (SEQIDNO:5) 1cgtcacgccccaggagaggcaataggaggccctggccctgccgacatggccaccgcagtc 61ccaacggcgcgctaggttggcgagatgaagaggagtcgcagtgccctgtccgtggcaggg 121accggggacgagaggtcgagggagacccccgaatccgaccgtgccaacatgctgggggcc 181gacctgcgtcgccctcgccgccgcctctcgtccggtcctggcctgggctgggcccagcct 241gagccctcggaccctggggtccctctgccgccacggcccaccaccctgccgctgctgatc 301ccaccgcggatttccatcaccagggccgagaacgacagcttcgaggcagagaatgggccg 361acaccatctcctggccgcagccccctggactcgcaggcgagcccaggactcgtgctgcac 421gccggggcggccaccagccagcgccgggagtccttcctgtaccgctcagacagcgactat 481gacatgtcacccaagaccatgtcccggaactcatcggtcaccagcgaggcgcacgctgaa 541gacctcatcgtaacaccatttgctcaggtgctggccagcctccggagcgtccgtagcaac 601ttctcactcctgaccaatgtgcccgttcccagtaacaagcggtccccgctgggcggcccc 661acccctgtctgcaaggccacgctgtcagaagaaacgtgtcagcagttggcccgggagact 721ctggaggagctggactggtgtctggagcagctggagaccatgcagacctatcgctctgtc 781agcgagatggcctcgcacaagttcaaaaggatgttgaaccgtgagctcacacacctgtca 841gaaatgagcaggtccggaaaccaggtctcagagtacatttccacaacattcctggacaaa 901cagaatgaagtggagatcccatcacccacgatgaaggaacgagaaaaacagcaagcgccg 961cgaccaagaccctcccagccgcccccgccccctgtaccacacttacagcccatgtcccaa 1021atcacagggttgaaaaagttgatgcatagtaacagcctgaacaactctaacattccccga 1081tttggggtgaagaccgatcaagaagagctcctggcccaagaactggagaacctgaacaag 1141tggggcctgaacatcttttgcgtgtcggattacgctggaggccgctcactcacctgcatc 1201atgtacatgatattccaggagcgggacctgctgaagaaattccgcatccctgtggacacg 1261atggtgacatacatgctgacgctggaggatcactaccacgctgacgtggcctaccataac 1321agcctgcacgcagctgacgtgctgcagtccacccacgtactgctggccacgcctgcacta 1381gatgcagtgttcacggacctggagattctcgccgccctcttcgcggctgccatccacgat 1441gtggatcaccctggggtctccaaccagttcctcatcaacaccaattcggagctggcgctc 1501atgtacaacgatgagtcggtgctcgagaatcaccacctggccgtgggcttcaagctgctg 1561caggaggacaactgcgacatcttccagaacctcagcaagcgccagcggcagagcctacgc 1621aagatggtcatcgacatggtgctggccacggacatgtccaagcacatgaccctcctggct 1681gacctgaagaccatggtggagaccaagaaagtgaccagctcaggggtcctcctgctagat 1741aactactccgaccgcatccaggtcctccggaacatggtgcactgtgccgacctcagcaac 1801cccaccaagccgctggagctgtaccgccagtggacagaccgcatcatggccgagttcttc 1861cagcagggtgaccgagagcgcgagcgtggcatggaaatcagccccatgtgtgacaagcac 1921actgcctccgtggagaagtctcaggtgggttttattgactacattgtgcacccattgtgg 1981gagacctgggcggaccttgtccacccagatgcccaggagatcttggacactttggaggac 2041aaccgggactggtactacagcgccatccggcagagcccatctccgccacccgaggaggag 2101tcaagggggccaggccacccacccctgcctgacaagttccagtttgagctgacgctggag 2161gaggaagaggaggaagaaatatcaatggcccagataccgtgcacagcccaagaggcattg 2221actgcgcagggattgtcaggagtcgaggaagctctggatgcaaccatagcctgggaggca 2281tccccggcccaggagtcgttggaagttatggcacaggaagcatccctggaggccgagctg 2341gaggcagtgtatttgacacagcaggcacagtccacaggcagtgcacctgtggctccggat 2401gagttctcgtcccgggaggaattcgtggttgctgtaagccacagcagcccctctgccctg 2461gctcttcaaagcccccttctccctgcttggaggaccctgtctgtttcagagcatgccccg 2521ggcctcccgggcctcccctccacggcggccgaggtggaggcccaacgagagcaccaggct 2581gccaagagggcttgcagtgcctgcgcagggacatttggggaggacacatccgcactccca 2641gctcctggtggcggggggtcaggtggagaccctacctgatccccagacctctgtccctgt 2701tcccctccactcctcccctcactcccctgctcccccgaccacctcctcctctgcctcaaa 2761gactcttgtcctcttgtccctcctgagaaaaaagaaaacgaaaagtggggtttttttctg 2821ttttctttttttcccctttccccctgcccccacccacggggcctttttttggaggtgggg 2881gctggggaatgaggggctgaggtcccggaagggattttatttttttgaattttaattgta 2941acatttttagaaaaagaacaaaaaaagaaaaaaaaaagaaagaaacacagcaactgtaga 3001tgctcctgttcctggttcccgctttccacttccaaatccctcccctcaccttcccccact 3061gccccccaagttccaggctcagtcttccagccgcctggggagtctctacctgggcccaag 3121caggtgtggggcctccttctgggcttttcttctgaatttagaggatttctagaacgtggt 3181caggaatagccattctaggcggggctggggccagggtggggggcagtcactgtgggaggt 3241cccagctccagcccccctctggtttgctgcctcctctcccctctaaaaaagtcttccgct 3301tgattttgcacaatcccggcgatactcctggcgatactgactagaaagtcagggagctgg 3361gggagctgttcactttaggatacgggggtggtatggaagggagcgttcacaccgccagcc 3421tcgggcctgggatttgaggagggccctagacctcctccactctccatcccctttcccttc 3481cactttgggttcactttgaattttctccgttttttggggcagtggctctgatccactcac 3541ccccccgccccccgccccacttctagctgcttctcctcttgtttctgccttaataattcc 3601cacggccacaggcaagggggttgcagtggccgcctgcaccttggatgaggcagggccagg 3661cgcccagaacccccatcctggccgcacccccctttccagggtcctccggaccccaccttc 3721cacactctgatcacagcccccctaccttttgccctaggaggaagcaataatggtgtatac 3781cctcattctcattcctgggcagcccttccttccaccctggcaccaaaataatttctcctc 3841catccgtaccttgcctagcctctccctctcccccagctagtccctgagcaatacggcaga 3901cagatgcaagaccatttttctccaagccatgggggactgtttggaaggaaagccccctct 3961ctccctcctcccctcgccctcggcctggttctgcagctggaccgacctcattcatcgcct 4021gccccctacccaattctgagcacacggtactgtagcccccagttcctccctagccttcca 4081tccctctgtccaccccagggggaggtaaccccgcactcacactcccttgatgctgtctgt 4141acagggttcatattttgtagcgaaagtcgtttttgtcccagccggcgatcggagtgggcc 4201ttttctttctttttgttcattctttacctttttttcttttctttctttcttttttgtaca 4261tactgtaaggttggtttgtaaattattctacggaggcaaaaagggaaaataaaaacttgc 4321ccttccctggctgacccagtcgggaaggtagggaaggaggtctcccgttgggagagtctc 4381tgttcctgctgtattatacaaactgtaccatagtcctgggaaaagggtggactcaccgct 4441gttgttttatgggaagtcgtgtcatcctaggggttggggctgggcagagcctgtcccctc 4501cccccttctccaggagccagggggtgactggagagacagacccacccccaagcagggctc 4561ctctccccagggtgagcacaggacctctgtaagctgcttgtgtattgtccactttgacga 4621tcagtcattcggtccgttgatcaataatccttcgatcttgtctccaattaaaccgaggct 4681ttcaccgataaaaaaaaaaaaaaa

[0949] An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a PDE4 protein. Non-coding regions (5 and 3 untranslated regions) are the 5 and 3 sequences that flank the coding region in a gene and are not translated into amino acids.

[0950] Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a PDE4 described herein. Antisense nucleic acids targeting a nucleic acid encoding a PDE4 can be designed using the software available at the Integrated DNA Technologies website.

[0951] An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

[0952] Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).

[0953] The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a PDE4 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

[0954] An antisense nucleic acid can be an -anomeric nucleic acid molecule. An -anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, -units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2-O-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

[0955] Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a PDE4 protein (e.g., specificity for a PDE4 mRNA, e.g., specificity for SEQ ID NO: 1, 2, 3, 4, or 5). Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a PDE4 mRNA can be designed based upon the nucleotide sequence of any of the PDE4 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a PDE4 mRNA (see, e.g., U.S. Pat. Nos. 4,987,071 and 5,116,742). Alternatively, a PDE4 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.

[0956] An inhibitor nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a PDE4 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the PDE4 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

[0957] In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Nat. Acad. Sci. U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.

[0958] PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

[0959] The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5-(4-methoxytrityl)amino-5-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5 end of DNA (Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5 PNA segment and a 3 DNA segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5 DNA segment and a 3 PNA segment (Peterser et al., Bioorganic Med. Chem. Lett. 5:1119-11124, 1975).

[0960] In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Nat. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc. Nat. Acad. Sci. U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

[0961] Another means by which expression of a PDE4 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a PDE4 polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3 terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).

[0962] RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Nat. Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Pat. No. 6,506,559 and US 2003/0056235, which are hereby incorporated by reference.

[0963] Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA or with plasmids engineered to make siRNA are routine in the art.

[0964] The siRNA molecules used to decrease expression of a PDE4 mRNA can vary in a number of ways. For example, they can include a 3 hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3 end, the 5 end, or both ends. For example, at least one strand of the RNA molecule can have a 3 overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4 or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

[0965] To further enhance the stability of the RNA duplexes, the 3 overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3 overhangs by 2-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing PDE4 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 1-5, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).

[0966] Non-limiting examples of siRNAs targeting PDE4 are described in Takakura et al., PLosOne 10(12):e0142981, 2015; Watanabe et al., Cell Signal 27(7):1517-1524, 2015; Suzuki et al., PLos One 11(7):e0158967, 2016; Kai et al., Mol. Ther. Nucl. Acids 6: 163-172, 2017). See, e.g., Cheng et al. Exp Ther Med 12(4): 2257-2264, 2016; Peter et al., J Immunol 178)8): 4820-4831; and Lynch et al. J Biolog Chem 280: 33178-33189. Additional examples of PDE4 inhibitory nucleic acids are described in U.S. Patent Application Publication Nos. 2010/0216703 and 2014/0171487, which are incorporated by reference in its entirety.

[0967] In some embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting PDE4 can be administered to a subject (e.g., a human subject) in need thereof.

[0968] In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5 or 3end of DNA or RNA.

[0969] Any of the inhibitor nucleic acids described herein can be formulated for administration to the gastrointestinal tract. See, e.g., the formulation methods described in US 2016/0090598 and Schoellhammer et al., Gastroenterology, doi: 10.1053/j.gastro.2017.01.002, 2017.

[0970] In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol. Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.

[0971] In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., J. Control Release 158(3): 362-370, 2012), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; Lin et al., Nanomedicine 9(1):105-120, 2014).

Additional Examples of Immune Modulators

[0972] An immune modulator as described herein can be an antibody or antigen-binding fragment, a nucleic acid (e.g., inhibitory nucleic acid), a small molecule, and a live biotherapeutic, such as a probiotic. In some embodiments, the immune modulator can be a drug or therapeutic used for the treatment of inflammatory bowel disease (IBD), for example, Crohn's Disease or Ulcerative Colitic (UC). Non-limiting immune modulators that useful for treating or preventing inflammatory bowel disease include substances that suppress cytokine production, down-regulate or suppress self-antigen expression, or mask MHC antigens. Non-limiting examples of immune modulators include, without limitation: CHST15 inhibitors (e.g., STNM01); IL-6 receptor inhibitora (e.g., tocilizumab); IL-12/IL-23 inhibitors (e.g., ustekinumab and brazikumab); integrin inhibitors (e.g., vedolizumab and natalizumab); JAK inhibitors (e.g., tofacitinib); SMAD7 inhibitors (e.g., Mongersen); IL-13 inhibitors; IL-1 receptor inhibitors; TLR agonists (e.g., Kappaproct); stem cells (e.g., Cx601); 2-amino-6-aryl-5-substituted pyrimidines (see U.S. Pat. No. 4,665,077); nonsteroidal anti-inflammatory drugs (NSAIDs); ganciclovir; tacrolimus; glucocorticoids such as Cortisol or aldosterone; anti-inflammatory agents such as a cyclooxygenase inhibitor; a 5-lipoxygenase inhibitor; or a leukotriene receptor antagonist; purine antagonists such as azathioprine or mycophenolate mofetil (MMF); alkylating agents such as cyclophosphamide; bromocryptine; danazol; dapsone; glutaraldehyde (which masks the MHC antigens, as described in U.S. Pat. No. 4,120,649); anti-idiotypic antibodies for MHC antigens and MHC fragments; cyclosporine; 6-mercaptopurine; steroids such as corticosteroids or glucocorticosteroids or glucocorticoid analogs, e.g., prednisone, methylprednisolone, including SOLU-MEDROL, methylprednisolone sodium succinate, and dexamethasone; dihydrofolate reductase inhibitors such as methotrexate (oral or subcutaneous); anti-malarial agents such as chloroquine and hydroxychloroquine; sulfasalazine; leflunomide; cytokine or cytokine receptor antibodies or antagonists including anti-interferon-alpha, -beta, or -gamma antibodies, anti-tumor necrosis factor(TNF)-alpha antibodies (infliximab (REMICADE) or adalimumab), anti-TNF-alpha immunoadhesin (etanercept), anti-TNF-beta antibodies, antiinterleukin-2 (IL-2) antibodies and anti-IL-2 receptor antibodies, and anti-interleukin-6 (IL-6) receptor antibodies and antagonists; anti-LFA-1 antibodies, including anti-CD 1 la and anti-CD 18 antibodies; anti-L3T4 antibodies; heterologous anti-lymphocyte globulin; pan-T antibodies, anti-CD3 or anti-CD4/CD4a antibodies; soluble peptide containing a LFA-3 binding domain (WO 90/08187 published Jul. 26, 1990); streptokinase; transforming growth factor-beta (TGF-beta); streptodomase; RNA or DNA from the host; FK506; RS-61443; chlorambucil; deoxyspergualin; rapamycin; T-cell receptor (Cohen et al, U.S. Pat. No. 5,114,721); T-cell receptor fragments (Offner et al, Science, 251: 430-432 (1991); WO 90/11294; Ianeway, Nature, 341: 482 (1989); and WO 91/01133); BAFF antagonists such as BAFF or BR3 antibodies or immunoadhesins and zTNF4 antagonists (for review, see Mackay and Mackay, Trends Immunol, 23: 113-5 (2002) and see also definition below); 10 biologic agents that interfere with T cell helper signals, such as anti-CD40 receptor or anti-CD40 ligand (CD 154), including blocking antibodies to CD40-CD40 ligand. (e.g., Durie et al, Science, 261 1328-30 (1993); Mohan et al, J. Immunol, 154: 1470-80 (1995)) and CTLA4-Ig (Finck et al, Science, 265: 1225-7 (1994)); and T-cell receptor antibodies (EP 340,109) such as T10B9. Non-limiting examples of agents also include the following: budenoside; epidermal growth factor; aminosalicylates; metronidazole; mesalamine; olsalazine; balsalazide; antioxidants; thromboxane inhibitors; IL-I receptor antagonists; anti-IL-I monoclonal antibodies; growth factors; elastase inhibitors; pyridinylimidazole compounds; TNF antagonists; IL-4, IL-10, IL-13 and/or TGF cytokines or agonists thereof (e.g., agonist antibodies); IL-11; glucuronide- or dextran-conjugated prodrugs of prednisolone, dexamethasone or budesonide; ICAM-I antisense phosphorothioate oligodeoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TPlO; T Cell Sciences, Inc.); slow-release mesalazine; antagonists of platelet activating factor (PAF); ciprofloxacin; and lignocaine.

[0973] Non-limiting examples of immune modulators that are useful for treating ulcerative colitis include sulfasalazine and related salicylate-containing drugs for mild cases and corticosteroid drugs for severe cases. Non-limiting examples of immune modulators that are useful for treating a liver disease or disorder (e.g., liver fibrosis or NASH) include: elafibranor (GFT 505; Genfit Corp.), obeticholic acid (OCA; Intercept Pharmaceuticals, Inc.), cenicriviroc (CVC; Allergan plc), selonsertib (formerly GS-4997; Gilead Sciences, Inc.), an anti-LOXL2 antibody (simtuzumab (formerly GS 6624; Gilead Sciences, Inc.)), GS-9450 (Gilead Sciences, Inc.), GS-9674 (Gilead Sciences, Inc.), GS-0976 (formerly NDI-010976; Gilead Sciences, Inc.), Emricasan (Conatus Pharmaceuticals, Inc.), Arachidyl-amido cholanoic acid (Aramchol; Galmed Pharmaceuticals Ltd.), AKN-083 (Allergan plc (Akarna Therapeutics Ltd.)), TGFTX4 (Genfit Corp.), TGFTX5 (Genfit Corp.), TGFTX1 (Genfit Corp.), a RoR agonist (e.g., LYC-55716; Lycera Corp.), an ileal bile acid transporter (iBAT) inhibitor (e.g., elobixibat, Albireo Pharma, Inc.; GSK2330672, GlaxoSmithKline plc; and A4250; Albireo Pharma, Inc.), stem cells, a CCR2 inhibitor, bardoxolone methyl (Reata Pharmaceuticals, Inc.), a bone morphogenetic protein-7 (BMP-7) mimetic (e.g., THR-123 (see, e.g., Sugimoto et al. (2012) Nature Medicine 18: 396-404)), an anti-TGF- antibody (e.g., fresolimumab; see also U.S. Pat. Nos. 7,527,791 and 8,383,780, incorporated herein by reference), pirfenidone (Esbriet, Genentech USA Inc.), an anti-integrin v6 antibody, an anti-connective tissue growth factor (CTGF) antibody (e.g., pamrevlumab; FibroGen Inc.), pentoxifylline, vascular endothelial growth factor (VEGF), a renin angiotensin aldosterone system (RAAS) inhibitor (e.g., a rennin inhibitor (e.g. pepstatin, CGP2928, aliskiren), or an ACE inhibitor (e.g., captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, fosinopril, and trandolapril)), thrombospondin, a statin, bardoxolone, a PDE5 inhibitor (e.g., sidenafil, vardenafil, and tadalafil), a NADPH oxidase-1 (NOX1) inhibitor (see, e.g., U.S. Publication No. 2011/0178082, incorporated herein by reference), a NADPH oxidase-4 (NOX4) inhibitor (see, e.g., U.S. Publication No. 2014/0323500, incorporated herein by reference), an ETA antagonist (e.g., sitaxentan, ambrisentan, atrasentan, BQ-123, and zibotentan), nintedanib (Boehringer Ingelheim), INT-767 (Intercept Pharmaceuticals, Inc.), VBY-376 (Virobay Inc.), PF-04634817(Pfizer), EXC 001 (Pfizer), GM-CT-01 (Galectin Therapeutics), GCS-100 (La Jolla Pharmaceuticals), hepatocyte growth factor mimetic (Refanalin; Angion Biomedica), SAR156597 (Sanofi), tralokinumab (AstraZeneca), pomalidomide (Celgene), STX-100 (Biogen IDEC), CC-930 (Celgene), anti-miR-21 (Regulus Therapeutics), PRM-151 (Promedior), BOT191 (BiOrion), Palomid 529 (Paloma Pharamaceuticals), IMD1041 (IMMD, Japan), serelaxin (Novartis), PEG-relaxin (Ambrx and Bristol-Myers Squibb), ANG-4011 (Angion Biomedica), FT011 (Fibrotech Therapeutics), pirfenidone (InterMune), F351 (pirfenidone derivative (GNI Pharma), vitamin E (e.g., tocotrienol (alpha, beta, gamma, and delta) and tocopherol (alpha, beta, gamma, and delta)), pentoxifylline, an insulin sensitizer (e.g., rosiglitazone and pioglitazone), cathepsin B inhibitor R-3020, etanercept and biosimilars thereof, peptides that block the activation of Fas (see, e.g., International Publication No. WO 2005/117940, incorporated herein by reference), caspase inhibitor VX-166, caspase inhibitor Z-VAD-fmk, fasudil, belnacasan (VX-765), and pralnacasan (VX-740).

[0974] Therapeutic agents that may be used for the treatment of the indications herein also include:

[0975] TNF inhibitors: tulinercept, DLX-105 (gel formulation);

[0976] IL-12/Il-23 inhibitors: AK-101;

[0977] IL-6R inhibitors: YSIL6, olokizumab (CDP-6038);

[0978] JAK inhibitors: PF-06700841, PF-06651600;

[0979] live biotherapeutics: Neuregulin 4; NN8555;

[0980] immune modulators: KHK-4083, GSK2618960, Toralizumab:

[0981] chemokines: GSK3050002 (previously known as KANAb071), E-6011, HGS-1025;

[0982] IL-1 inhibitors: K(D)PT;

[0983] IL-10 inhibitors: RG-7880;

[0984] CHST15 inhibitors: SB-012:

[0985] TLR agonists: BL-7040; EN-101; Monarsen.

[0986] In some embodiments, an immune modulator can decrease the activity and/or the level in a mammalian cell of its target receptor, such as TNF, IL-12/IL-23, IL-6R, JAK, a chemokine, IL-1, IL-10, CHST15, or TLR. In some embodiments, a immune modulator can decrease (e.g., by about 1% to about 99%, by about 1% to about 95%, by about 1% to about 90%, by about 1% to about 85%, by about 1% to about 80%, by about 1% to about 75%, by about 1% to about 70%, by about 1% to about 65%, by about 1% to about 60%, by about 1% to about 55%, by about 1% to about 50%, by about 1% to about 45%, by about 1% to about 40%, by about 1% to about 35%, by about 1% to about 30%, by about 1% to about 25%, by about 1% to about 20%, by about 1% to about 20%, by about 1% to about 15%, by about 1% to about 10%, by about 1% to about 5%, by about 5% to about 99%, by about 5% to about 90%, by about 5% to about 85%, by about 5% to about 80%, by about 5% to about 75%, by about 5% to about 70%, by about 5% to about 65%, by about 5% to about 60%, by about 5% to about 55%, by about 5% to about 50%, by about 5% to about 45%, by about 5% to about 40%, by about 5% to about 35%, by about 5% to about 30%, by about 5% to about 25%, by about 5% to about 20%, by about 5% to about 15%, by about 5% to about 10%, by about 10% to about 99%, about 10% to about 95%, about 10% to about 90%, about 10% to about 85%, by about 10% to about 80%, by about 10% to about 75%, by about 10% to about 70%, by about 10% to about 65%, by about 10% to about 60%, by about 10% to about 55%, by about 10% to about 50%, by about 10% to about 45%, by about 10% to about 40%, by about 10% to about 35%, by about 10% to about 30%, by about 10% to about 25%, by about 10% to about 20%, by about 10% to about 15%, by about 15% to about 99%, by about 15% to about 95%, by about 15% to about 90%, by about 15% to about 85%, by about 15% to about 80%, by about 15% to about 75%, by about 15% to about 70%, by about 15% to about 65%, by about 15% to about 60%, by about 15% to about 55%, by about 15% to about 50%, by about 15% to about 45%, by about 15% to about 40%, by about 15% to about 35%, by about 15% to about 30%, by about 15% to about 25%, by about 15% to about 20%, by about 20% to about 99%, by about 20% to about 95%, by about 20% to about 90%, by about 20% to about 85%, by about 20% to about 80%, by about 20% to about 75%, by about 20% to about 70%, by about 20% to about 65%, by about 20% to about 60%, by about 20% to about 55%, by about 20% to about 50%, by about 20% to about 45%, by about 20% to about 40%, by about 20% to about 35%, by about 20% to about 30%, by about 20% to about 25%, by about 25% to about 99%, about 25% to about 95%, by about 25% to about 90%, by about 25% to about 85%, by about 25% to about 80%, by about 25% to about 75%, by about 25% to about 70%, by about 25% to about 65%, by about 25% to about 60%, by about 25% to about 55%, by about 25% to about 50%, by about 25% to about 45%, by about 25% to about 40%, by about 25% to about 35%, by about 25% to about 30%, by about 30% to about 99%, by about 30% to about 95%, by about 30% to about 90%, by about 30% to about 85%, by about 30% to about 80%, by about 30% to about 75%, by about 30% to about 70%, by about 30% to about 65%, by about 30% to about 60%, by about 30% to about 55%, by about 30% to about 50%, by about 30% to about 45%, by about 30% to about 40%, by about 30% to about 35%, by about 35% to about 99%, by about 35% to about 95%, by about 35% to about 90%, by about 35% to about 85%, by about 35% to about 80%, by about 35% to about 75%, by about 35% to about 70%, by about 35% to about 65%, by about 35% to about 60%, by about 35% to about 55%, by about 35% to about 50%, by about 35% to about 45%, by about 35% to about 40%, by about 40% to about 99%, by about 40% to about 95%, by about 40% to about 90%, by about 40% to about 85%, by about 40% to about 80%, by about 40% to about 75%, by about 40% to about 70%, by about 40% to about 65%, by about 40% to about 60%, by about 40% to about 55%, by about 40% to about 50%, by about 40% to about 45%, by about 45% to about 99%, by about 45% to about 95%, by about 45% to about 90%, by about 45% to about 85%, by about 45% to about 80%, by about 45% to about 75%, by about 45% to about 70%, by about 45% to about 65%, by about 45% to about 60%, by about 45% to about 55%, by about 45% to about 50%, by about 50% to about 99%, by about 50% to about 95%, by about 50% to about 90%, by about 50% to about 85%, by about 50% to about 80%, by about 50% to about 75%, by about 50% to about 70%, by about 50% to about 65%, by about 50% to about 60%, by about 50% to about 55%, by about 55% to about 99%, by about 55% to about 95%, by about 55% to about 90%, by about 55% to about 85%, by about 55% to about 80%, by about 55% to about 75%, by about 55% to about 70%, by about 55% to about 65%, by about 55% to about 60%, by about 60% to about 99%, by about 60% to about 95%, by about 60% to about 90%, by about 60% to about 85%, by about 60% to about 80%, by about 60% to about 75%, by about 60% to about 70%, by about 60% to about 65%, by about 65% to about 99%, by about 65% to about 95%, by about 65% to about 90%, by about 65% to about 85%, by about 65% to about 80%, by about 65% to about 75%, by about 65% to about 70%, by about 70% to about 99%, by about 70% to about 95%, by about 70% to about 90%, by about 70% to about 85%, by about 70% to about 80%, by about 70% to about 75%, by about 75% to about 99%, by about 75% to about 95%, by about 75% to about 90%, by about 75% to about 85%, by about 75% to about 80%, by about 80% to about 99%, by about 80% to about 95%, by about 80% to about 90%, by about 80% to about 85%, by about 85% to about 99%, by about 85% to about 95%, by about 85% to about 90%, by about 90% to about 99%, by about 90% to about 95%, or by about 95% to about 99%) in the level of PDE4 protein in a mammalian cell contacted with the agent, e.g., as compared to the level of PDE4 protein in the same mammalian cell not contacted with the agent.

[0987] In some embodiments, a immune modulator can inhibit PDE4 activity with an IC.sub.50 of about 1 pM to about 100 M, about 1 pM to about 95 M, about 1 pM to about 90 M, about 1 pM to about 85 M, about 1 pM to about 80 M, about 1 pM to about 75 M, about 1 pM to about 70 M, about 1 pM to about 65 M, about 1 pM to about 60 M, about 1 pM to about 55 M, about 1 pM to about 50 M, about 1 pM to about 45 M, about 1 pM to about 40 M, about 1 pM to about 35 M, about 1 pM to about 30 M, about 1 pM to about 25 M, about 1 pM to about 20 M, about 1 pM to about 15 M, about 1 pM to about 10 M, about 1 pM to about 5 M, about 1 pM to about 1 M, about 1 pM to about 900 nM, about 1 pM to about 800 nM, about 1 pM to about 700 nM, about 1 pM to about 600 nM, about 1 pM to about 500 nM, about 1 pM to about 400 nM, about 1 pM to about 300 nM, about 1 pM to about 200 nM, about 1 pM to about 100 nM, about 1 pM to about 50 nM, about 1 pM to about 1 nM, about 1 pM to about 800 pM, about 1 pM to about 600 pM, about 1 pM to about 400 pM, about 1 pM to about 200 pM, about 200 pM to about 100 pM, about 200 pM to about 95 M, about 200 pM to about 90 M, about 200 pM to about 85 M, about 200 pM to about 80 M, about 200 pM to about 75 M, about 200 pM to about 70 M, about 200 pM to about 65 M, about 200 pM to about 60 M, about 200 pM to about 55 M, about 200 pM to about 50 M, about 200 pM to about 45 M, about 200 pM to about 40 M, about 200 pM to about 35 M, about 200 pM to about 30 M, about 200 pM to about 25 M, about 200 pM to about 20 M, about 200 pM to about 15 M, about 200 pM to about 10 M, about 200 pM to about 5 M, about 200 pM to about 1 M, about 200 pM to about 900 nM, about 200 pM to about 800 nM, about 200 pM to about 700 nM, about 200 pM to about 600 nM, about 200 pM to about 500 nM, about 200 pM to about 400 nM, about 200 pM to about 300 nM, about 200 pM to about 200 nM, about 200 pM to about 100 nM, about 200 pM to about 50 nM, about 200 pM to about 1 nM, about 200 pM to about 800 pM, about 200 pM to about 600 pM, about 200 pM to about 400 pM, about 400 pM to about 100 M, about 400 pM to about 95 M, about 400 pM to about 90 M, about 400 pM to about 85 M, about 400 pM to about 80 M, about 400 pM to about 75 M, about 400 pM to about 70 M, about 400 pM to about 65 M, about 400 pM to about 60 M, about 400 pM to about 55 M, about 400 pM to about 50 M, about 400 pM to about 45 M, about 400 pM to about 40 M, about 400 pM to about 35 M, about 400 pM to about 30 M, about 400 pM to about 25 M, about 400 pM to about 20 M, about 400 pM to about 15 M, about 400 pM to about 10 M, about 400 pM to about 5 M, about 400 pM to about 1 M, about 400 pM to about 900 nM, about 400 pM to about 800 nM, about 400 pM to about 700 nM, about 400 pM to about 600 nM, about 400 pM to about 500 nM, about 400 pM to about 400 nM, about 400 pM to about 300 nM, about 400 pM to about 200 nM, about 400 pM to about 100 nM, about 400 pM to about 50 nM, about 400 pM to about 1 nM, about 400 pM to about 800 pM, 400 pM to about 600 pM, about 600 pM to about 100 M, about 600 pM to about 95 M, about 600 pM to about 90 M, about 600 pM to about 85 M, about 600 pM to about 80 M, about 600 pM to about 75 M, about 600 pM to about 70 M, about 600 pM to about 65 M, about 600 pM to about 60 M, about 600 pM to about 55 M, about 600 pM to about 50 M, about 600 pM to about 45 M, about 600 pM to about 40 M, about 600 pM to about 35 M, about 600 pM to about 30 M, about 600 pM to about 25 M, about 600 pM to about 20 M, about 600 pM to about 15 M, about 600 pM to about 10 M, about 600 pM to about 5 M, about 600 pM to about 1 M, about 600 pM to about 900 nM, about 600 pM to about 800 nM, about 600 pM to about 700 nM, about 600 pM to about 600 nM, about 600 pM to about 500 nM, about 600 pM to about 400 nM, about 600 pM to about 300 nM, about 600 pM to about 200 nM, about 600 pM to about 100 nM, about 600 pM to about 50 nM, about 600 pM to about 1 nM, about 600 pM to about 800 pM, about 800 pM to about 100 M, about 800 pM to about 95 M, about 800 pM to about 90 M, about 800 pM to about 85 M, about 800 pM to about 80 M, about 800 pM to about 75 M, about 800 pM to about 70 M, about 800 pM to about 65 M, about 800 pM to about 60 M, about 800 pM to about 55 M, about 800 pM to about 50 M, about 800 pM to about 45 M, about 800 pM to about 40 M, about 800 pM to about 35 M, about 800 pM to about 30 M, about 800 pM to about 25 M, about 800 pM to about 20 M, about 800 pM to about 15 M, about 800 pM to about 10 M, about 800 pM to about 5 M, about 800 pM to about 1 M, about 800 pM to about 900 nM, about 800 pM to about 800 nM, about 800 pM to about 700 nM, about 800 pM to about 600 nM, about 800 pM to about 500 nM, about 800 pM to about 400 nM, about 800 pM to about 300 nM, about 800 pM to about 200 nM, about 800 pM to about 100 nM, about 800 pM to about 50 nM, about 800 pM to about 1 nM, about 1 nM to about 100 M, about 1 nM to about 95 M, about 1 nM to about 90 M, about 1 nM to about 85 M, about 1 nM to about 80 M, about 1 nM to about 75 M, about 1 nM to about 70 M, about 1 nM to about 65 M, about 1 nM to about 60 M, about 1 nM to about 55 M, about 1 nM to about 50 M, about 1 nM to about 45 M, about 1 nM to about 40 M, about 1 nM to about 35 M, about 1 nM to about 30 M, about 1 nM to about 25 M, about 1 nM to about 20 M, about 1 nM to about 15 M, about 1 nM to about 10 M, about 1 nM to about 5 M, about 1 nM to about 1 M, about 1 nM to about 900 nM, about 1 nM to about 800 nM, about 1 nM to about 700 nM, about 1 nM to about 600 nM, about 1 nM to about 500 nM, about 1 nM to about 400 nM, about 1 nM to about 300 nM, about 1 nM to about 200 nM, about 1 nM to about 100 nM, about 1 nM to about 50 nM, about 50 nM to about 100 M, about 50 nM to about 95 M, about 50 nM to about 90 M, about 50 nM to about 85 M, about 50 nM to about 80 M, about 50 nM to about 75 M, about 50 nM to about 70 M, about 50 nM to about 65 M, about 50 nM to about 60 M, about 50 nM to about 55 M, about 50 nM to about 50 M, about 50 nM to about 45 M, about 50 nM to about 40 M, about 50 nM to about 35 M, about 50 nM to about 30 M, about 50 nM to about 25 M, about 50 nM to about 20 M, about 50 nM to about 15 M, about 50 nM to about 10 M, about 50 nM to about 5 M, about 50 nM to about 1 M, about 50 nM to about 900 nM, about 50 nM to about 800 nM, about 50 nM to about 700 nM, about 50 nM to about 600 nM, about 50 nM to about 500 nM, about 50 nM to about 400 nM, about 50 nM to about 300 nM, about 50 nM to about 200 nM, about 50 nM to about 100 nM, about 100 nM to about 100 M, about 100 nM to about 95 M, about 100 nM to about 90 M, about 100 nM to about 85 M, about 100 nM to about 80 M, about 100 nM to about 75 M, about 100 nM to about 70 M, about 100 nM to about 65 M, about 100 nM to about 60 M, about 100 nM to about 55 M, about 100 nM to about 50 M, about 100 nM to about 45 M, about 100 nM to about 40 M, about 100 nM to about 35 M, about 100 nM to about 30 M, about 100 nM to about 25 M, about 100 nM to about 20 M, about 100 nM to about 15 M, about 100 nM to about 10 M, about 100 nM to about 5 M, about 100 nM to about 1 M, about 100 nM to about 900 nM, about 100 nM to about 800 nM, about 100 nM to about 700 nM, about 100 nM to about 600 nM, about 100 nM to about 500 nM, about 100 nM to about 400 nM, about 100 nM to about 300 nM, about 100 nM to about 200 nM, about 200 nM to about 100 M, about 200 nM to about 95 M, about 200 nM to about 90 M, about 200 nM to about 85 M, about 200 nM to about 80 M, about 200 nM to about 75 M, about 200 nM to about 70 M, about 200 nM to about 65 M, about 200 nM to about 60 M, about 200 nM to about 55 M, about 200 nM to about 50 M, about 200 nM to about 45 M, about 200 nM to about 40 M, about 200 nM to about 35 M, about 200 nM to about 30 M, about 200 nM to about 25 M, about 200 nM to about 20 M, about 200 nM to about 15 M, about 200 nM to about 10 M, about 200 nM to about 5 M, about 200 nM to about 1 M, about 200 nM to about 900 nM, about 200 nM to about 800 nM, about 200 nM to about 700 nM, about 200 nM to about 600 nM, about 200 nM to about 500 nM, about 200 nM to about 400 nM, about 200 nM to about 300 nM, about 300 nM to about 100 M, about 300 nM to about 95 M, about 300 nM to about 90 M, about 300 nM to about 85 M, about 300 nM to about 80 M, about 300 nM to about 75 M, about 300 nM to about 70 M, about 300 nM to about 65 M, about 300 nM to about 60 M, about 300 nM to about 55 M, about 300 nM to about 50 M, about 300 nM to about 45 M, about 300 nM to about 40 M, about 300 nM to about 35 M, about 300 nM to about 30 M, about 300 nM to about 25 M, about 300 nM to about 20 M, about 300 nM to about 15 M, about 300 nM to about 10 M, about 300 nM to about 5 M, about 300 nM to about 1 M, about 300 nM to about 900 nM, about 300 nM to about 800 nM, about 300 nM to about 700 nM, about 300 nM to about 600 nM, about 300 nM to about 500 nM, about 300 nM to about 400 nM, about 400 nM to about 100 M, about 400 nM to about 95 M, about 400 nM to about 90 M, about 400 nM to about 85 M, about 400 nM to about 80 M, about 400 nM to about 75 M, about 400 nM to about 70 M, about 400 nM to about 65 M, about 400 nM to about 60 M, about 400 nM to about 55 M, about 400 nM to about 50 M, about 400 nM to about 45 M, about 400 nM to about 40 M, about 400 nM to about 35 M, about 400 nM to about 30 M, about 400 nM to about 25 M, about 400 nM to about 20 M, about 400 nM to about 15 M, about 400 nM to about 10 M, about 400 nM to about 5 M, about 400 nM to about 1 M, about 400 nM to about 900 nM, about 400 nM to about 800 nM, about 400 nM to about 700 nM, about 400 nM to about 600 nM, about 400 nM to about 500 nM, about 500 nM to about 100 M, about 500 nM to about 95 M, about 500 nM to about 90 M, about 500 nM to about 85 M, about 500 nM to about 80 M, about 500 nM to about 75 M, about 500 nM to about 70 M, about 500 nM to about 65 M, about 500 nM to about 60 M, about 500 nM to about 55 M, about 500 nM to about 50 M, about 500 nM to about 45 M, about 500 nM to about 40 M, about 500 nM to about 35 M, about 500 nM to about 30 M, about 500 nM to about 25 M, about 500 nM to about 20 M, about 500 nM to about 15 M, about 500 nM to about 10 M, about 500 nM to about 5 M, about 500 nM to about 1 M, about 500 nM to about 900 nM, about 500 nM to about 800 nM, about 500 nM to about 700 nM, about 500 nM to about 600 nM, about 600 nM to about 100 M, about 600 nM to about 95 M, about 600 nM to about 90 M, about 600 nM to about 85 M, about 600 nM to about 80 M, about 600 nM to about 75 M, about 600 nM to about 70 M, about 600 nM to about 65 M, about 600 nM to about 60 M, about 600 nM to about 55 M, about 600 nM to about 50 M, about 600 nM to about 45 M, about 600 nM to about 40 M, about 600 nM to about 35 M, about 600 nM to about 30 M, about 600 nM to about 25 M, about 600 nM to about 20 M, about 600 nM to about 15 M, about 600 nM to about 10 M, about 600 nM to about 5 M, about 600 nM to about 1 M, about 600 nM to about 900 nM, about 600 nM to about 800 nM, about 600 nM to about 700 nM, about 700 nM to about 100 M, about 700 nM to about 95 M, about 700 nM to about 90 M, about 700 nM to about 85 M, about 700 nM to about 80 M, about 700 nM to about 75 M, about 700 nM to about 70 M, about 700 nM to about 65 M, about 700 nM to about 60 M, about 700 nM to about 55 M, about 700 nM to about 50 M, about 700 nM to about 45 M, about 700 nM to about 40 M, about 700 nM to about 35 M, about 700 nM to about 30 M, about 700 nM to about 25 M, about 700 nM to about 20 M, about 700 nM to about 15 M, about 700 nM to about 10 M, about 700 nM to about 5 M, about 700 nM to about 1 M, about 700 nM to about 900 nM, about 700 nM to about 800 nM, about 800 nM to about 100 M, about 800 nM to about 95 M, about 800 nM to about 90 M, about 800 nM to about 85 M, about 800 nM to about 80 M, about 800 nM to about 75 M, about 800 nM to about 70 M, about 800 nM to about 65 M, about 800 nM to about 60 M, about 800 nM to about 55 M, about 800 nM to about 50 M, about 800 nM to about 45 M, about 800 nM to about 40 M, about 800 nM to about 35 M, about 800 nM to about 30 M, about 800 nM to about 25 M, about 800 nM to about 20 M, about 800 nM to about 15 M, about 800 nM to about 10 M, about 800 nM to about 5 M, about 800 nM to about 1 M, about 800 nM to about 900 nM, about 900 nM to about 100 M, about 900 nM to about 95 M, about 900 nM to about 90 M, about 900 nM to about 85 M, about 900 nM to about 80 M, about 900 nM to about 75 M, about 900 nM to about 70 M, about 900 nM to about 65 M, about 900 nM to about 60 M, about 900 nM to about 55 M, about 900 nM to about 50 M, about 900 nM to about 45 M, about 900 nM to about 40 M, about 900 nM to about 35 M, about 900 nM to about 30 M, about 900 nM to about 25 M, about 900 nM to about 20 M, about 900 nM to about 15 M, about 900 nM to about 10 M, about 900 nM to about 5 M, about 900 nM to about 1 M, about 1 M to about 100 M, about 1 M to about 95 M, about 1 M to about 90 M, about 1 M to about 85 M, about 1 M to about 80 M, about 1 M to about 75 M, about 1 M to about 70 M, about 1 M to about 65 M, about 1 M to about 60 M, about 1 M to about 55 M, about 1 M to about 50 M, about 1 M to about 45 M, about 1 M to about 40 M, about 1 M to about 35 M, about 1 M to about 30 M, about 1 M to about 25 M, about 1 M to about 20 M, about 1 M to about 15 M, about 1 M to about 10 M, about 1 M to about 5 M, about 5 M to about 100 M, about 5 M to about 95 M, about 5 M to about 90 M, about 5 M to about 85 M, about 5 M to about 80 M, about 5 M to about 75 M, about 5 M to about 70 M, about 5 M to about 65 M, about 5 M to about 60 M, about 5 M to about 55 M, about 5 M to about 50 M, about 5 M to about 45 M, about 5 M to about 40 M, about 5 M to about 35 M, about 5 M to about 30 M, about 5 M to about 25 M, about 5 M to about 20 M, about 5 M to about 15 M, about 5 M to about 10 M, about 10 M to about 100 M, about 10 M to about 95 M, about 10 M to about 90 M, about 10 M to about 85 M, about 10 M to about 80 M, about 10 M to about 75 M, about 10 M to about 70 M, about 10 M to about 65 M, about 10 M to about 60 M, about 10 M to about 55 M, about 10 M to about 50 M, about 10 M to about 45 M, about 10 M to about 40 M, about 10 M to about 35 M, about 10 M to about 30 M, about 10 M to about 25 M, about 10 M to about 20 M, about 10 M to about 15 M, about 15 M to about 100 M, about 15 M to about 95 M, about 15 M to about 90 M, about 15 M to about 85 M, about 15 M to about 80 M, about 15 M to about 75 M, about 15 M to about 70 M, about 15 M to about 65 M, about 15 M to about 60 M, about 15 M to about 55 M, about 15 M to about 50 M, about 15 M to about 45 M, about 15 M to about 40 M, about 15 M to about 35 M, about 15 M to about 30 M, about 15 M to about 25 M, about 15 M to about 20 M, about 20 M to about 100 M, about 20 M to about 95 M, about 20 M to about 90 M, about 20 M to about 85 M, about 20 M to about 80 M, about 20 M to about 75 M, about 20 M to about 70 M, about 20 M to about 65 M, about 20 M to about 60 M, about 20 M to about 55 M, about 20 M to about 50 M, about 20 M to about 45 M, about 20 M to about 40 M, about 20 M to about 35 M, about 20 M to about 30 M, about 20 M to about 25 M, about 25 M to about 100 M, about 25 M to about 95 M, about 25 M to about 90 M, about 25 M to about 85 M, about 25 M to about 80 M, about 25 M to about 75 M, about 25 M to about 70 M, about 25 M to about 65 M, about 25 M to about 60 M, about 25 M to about 55 M, about 25 M to about 50 M, about 25 M to about 45 M, about 25 M to about 40 M, about 25 M to about 35 M, about 25 M to about 30 M, about 30 M to about 100 M, about 30 M to about 95 M, about 30 M to about 90 M, about 30 M to about 85 M, about 30 M to about 80 M, about 30 M to about 75 M, about 30 M to about 70 M, about 30 M to about 65 M, about 30 M to about 60 M, about 30 M to about 55 M, about 30 M to about 50 M, about 30 M to about 45 M, about 30 M to about 40 M, about 30 M to about 35 M, about 35 M to about 100 M, about 35 M to about 95 M, about 35 M to about 90 M, about 35 M to about 85 M, about 35 M to about 80 M, about 35 M to about 75 M, about 35 M to about 70 M, about 35 M to about 65 M, about 35 M to about 60 M, about 35 M to about 55 M, about 35 M to about 50 M, about 35 M to about 45 M, about 35 M to about 40 M, about 40 M to about 100 M, about 40 M to about 95 M, about 40 M to about 90 M, about 40 M to about 85 M, about 40 M to about 80 M, about 40 M to about 75 M, about 40 M to about 70 M, about 40 M to about 65 M, about 40 M to about 60 M, about 40 M to about 55 M, about 40 M to about 50 M, about 40 M to about 45 M, about 45 M to about 100 M, about 45 M to about 95 M, about 45 M to about 90 M, about 45 M to about 85 M, about 45 M to about 80 M, about 45 M to about 75 M, about 45 M to about 70 M, about 45 M to about 65 M, about 45 M to about 60 M, about 45 M to about 55 M, about 45 M to about 50 M, about 50 M to about 100 M, about 50 M to about 95 M, about 50 M to about 90 M, about 50 M to about 85 M, about 50 M to about 80 M, about 50 M to about 75 M, about 50 M to about 70 M, about 50 M to about 65 M, about 50 M to about 60 M, about 50 M to about 55 M, about 55 M to about 100 M, about 55 M to about 95 M, about 55 M to about 90 M, about 55 M to about 85 M, about 55 M to about 80 M, about 55 M to about 75 M, about 55 M to about 70 M, about 55 M to about 65 M, about 55 M to about 60 M, about 60 M to about 100 M, about 60 M to about 95 M, about 60 M to about 90 M, about 60 M to about 85 M, about 60 M to about 80 M, about 60 M to about 75 M, about 60 M to about 70 M, about 60 M to about 65 M, about 65 M to about 100 M, about 65 M to about 95 M, about 65 M to about 90 M, about 65 M to about 85 M, about 65 M to about 80 M, about 65 M to about 75 M, about 65 M to about 70 M, about 70 M to about 100 M, about 70 M to about 95 M, about 70 M to about 90 M, about 70 M to about 85 M, about 70 M to about 80 M, about 70 M to about 75 M, about 75 M to about 100 M, about 75 M to about 95 M, about 75 M to about 90 M, about 75 M to about 85 M, about 75 M to about 80 M, about 80 M to about 100 M, about 80 M to about 95 M, about 80 M to about 90 M, about 80 M to about 85 M, about 85 M to about 100 M, about 85 M to about 95 M, about 85 M to about 90 M, about 90 M to about 100 M, about 90 M to about 95 M, or about 95 M to about 100 M.

Exemplary Embodiments

Endoscopes, Ingestible Devices, and Reservoirs Containing the Drug

[0988] The GI tract can be imaged using endoscopes, or more recently ingestible devices that are swallowed.

[0989] The technology behind standard colonoscopy consists of a long, semi-rigid insertion tube with a steerable tip (stiff if compared to the colon), which is pushed by the physician from the outside. However, invasiveness, patient discomfort, fear of pain, and -more often than notthe need for conscious sedation limit the take-up of screening colonoscopy. Diagnosis and treatment in the GI tract are dominated by the use of flexible endoscopes. A few large companies, namely Olympus Medical Systems Co. (Tokyo, Japan), Pentax Medical Co. (Montvale, N.J., USA), Fujinon, Inc. (Wayne, N.J., USA) and Karl Storz GmbH & Co. KG (Tuttlingen, Germany), cover the majority of the market in flexible GI endoscopy.

[0990] Endoscopes may comprise a catheter. As an example, the catheter may be a spray catheter. As an example, a spray catheter may be used to deliver dyes for diagnostic purposes. As an example, a spray catheter may be used to deliver a therapeutic agent at an intended site in the GI tract. For example, the Olypmus PW-205V is a ready-to-use spray catheter that enables efficient spraying for maximal differentiation of tissue structures during endoscopy, but may also be used to deliver drugs.

[0991] Endoscopes may comprise a catheter. As an example, the catheter may be a spray catheter. As an example, a spray catheter may be used to deliver dyes for diagnostic purposes. As an example, a spray catheter may be used to deliver a therapeutic agent at the site of disease in the GI tract. For example, the Olypmus PW-205V is a ready-to-use spray catheter that enables efficient spraying for maximal differentiation of tissue structures during endoscopy, but may also be used to deliver drugs diseased tissue.

[0992] In a review of robotic endoscopic capsules, Journal of Micro-Bio Robotics 11.1-4 (2016): 1-18, Ciuti et al. state that progress in micro-electromechanical systems (MEMS) technologies have led to the development of new endoscopic capsules with enhanced diagnostic capabilities, in addition to traditional visualization of mucosa (embedding, e.g. pressure, pH, blood detection and temperature sensors).

[0993] Endoscopic capsules, however, do not have the capability of accurately locating a site autonomously. They require doctor oversight over a period of hours in order to manually determine the location. Autonomous ingestible devices are advantageous in that regard.

[0994] Ingestible devices are also advantageous over spray catheters in that they are less invasive, thereby allowing for regular dosing more frequently than spray catheters. Another advantage of ingestible devices is the greater ease with which they can access, relative to a catheter, certain sections of the GI tract such as the ascending colon, the cecum, and all portions of the small intestine.

Methods and Mechanisms for Localization

[0995] In addition to, or as an alternative, to directly visualizing the GI tract, one or more different mechanisms can be used to determine the location of an ingestible device within the GI tract. Various implementations may be used for localization of ingestible devices within the GI tract. For example, certain implementations can include one or more electromagnetic sensor coils, magnetic fields, electromagnetic waves, electric potential values, ultrasound positioning systems, gamma scintigraphy techniques or other radio-tracker technology have been described by others. Alternatively, imaging can be used to localize, for example, using anatomical landmarks or more complex algorithms for 3D reconstruction based on multiple images. Other technologies rely on radio frequency, which relies on sensors placed externally on the body to receive the strength of signals emitted by the capsule. Ingestible devices may also be localized based on reflected light in the medium surrounding the device; pH; temperature; time following ingestion; and/or acoustic signals.

[0996] The disclosure provides an ingestible device, as well as related systems and methods that provide for determining the position of the ingestible device within the GI tract of a subject with very high accuracy. In some embodiments, the ingestible device can autonomously determine its position within the GI tract of the subject.

[0997] Typically, the ingestible device includes one or more processing devices, and one more machine readable hardware storage devices. In some embodiments, the one or more machine readable hardware storage devices store instructions that are executable by the one or more processing devices to determine the location of the ingestible device in a portion of a GI tract of the subject. In certain embodiments, the one or more machine readable hardware storage devices store instructions that are executable by the one or more processing devices to transmit data to an external device (e.g., abase station external to the subject, such as abase station carried on an article worn by the subject) capable of implementing the data to determine the location of the device within the GI tract of the subject.

[0998] In some embodiments, the location of the ingestible device within the GI tract of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. In some embodiments, the location of the ingestible device within the GI tract of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. In such embodiments, the portion of the GI tract of the subject can include, for example, the esophagus, the stomach, duodenum, the jejunum, and/or the terminal ileum, cecum and colon. An exemplary and non-limiting embodiment is provided below in Example 14.

[0999] In certain embodiments, the location of the ingestible device within the esophagus of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. An exemplary and non-limiting embodiment is provided below in Example 14.

[1000] In some embodiments, the location of the ingestible device within the stomach of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. An exemplary and non-limiting embodiment is provided below in Example 14.

[1001] In certain embodiments, the location of the ingestible device within the duodenum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. An exemplary and non-limiting embodiment is provided below in Example 14.

[1002] In some embodiments, the location of the ingestible device within the jejunum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. An exemplary and non-limiting embodiment is provided below in Example 14.

[1003] In certain embodiments, the location of the ingestible device within the terminal ileum, cecum and colon of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1004] In some embodiments, the location of the ingestible device within the cecum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%. An exemplary and non-limiting embodiment is provided below in Example 14. In such embodiments, the portion of the portion of the GI tract of the subject can include, for example, the esophagus, the stomach, duodenum, the jejunum, and/or the terminal ileum, cecum and colon.

[1005] In certain embodiments, the location of the ingestible device within the esophagus of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1006] In some embodiments, the location of the ingestible device within the stomach of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1007] In certain embodiments, the location of the ingestible device within the duodenum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1008] In some embodiments, the location of the ingestible device within the jejunum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1009] In certain embodiments, the location of the ingestible device within the terminal ileum, cecum and colon of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1010] In some embodiments, the location of the ingestible device within the cecum of the subject can be determined to an accuracy of at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, 100%.

[1011] As used herein, the term reflectance refers to a value derived from light emitted by the device, reflected back to the device, and received by a detector in or on the device. For example, in some embodiments this refers to light emitted by the device, wherein a portion of the light is reflected by a surface external to the device, and the light is received by a detector located in or on the device.

[1012] As used herein, the term illumination refers to any electromagnetic emission. In some embodiments, an illumination may be within the range of Infrared Light (IR), the visible spectrum and ultraviolet light (UV), and an illumination may have a majority of its power centered at a particular wavelength in the range of 100 nm to 1000 nm. In some embodiments, it may be advantageous to use an illumination with a majority of its power limited to one of the infrared (750 nm-1000 nm), red (600 nm-750 nm), green (495 nm-600 nm), blue (400 nm-495 nm), or ultraviolet (100 nm-400 nm) spectrums. In some embodiments a plurality of illuminations with different wavelengths may be used. For illustrative purposes, the embodiments described herein may refer to the use of green or blue spectrums of light. However, it is understood that these embodiments may use any suitable light having a wavelength that is substantially or approximately within the green or blue spectra defined above, and the localization systems and methods described herein may use any suitable spectra of light.

[1013] Referring now to FIG. 1, shown therein is a view of an example embodiment of an ingestible device 100, which may be used to identify a location within a gastrointestinal (GI) tract. In some embodiments, ingestible device 100 may be configured to autonomously determine whether it is located in the stomach, a particular portion of the small intestine such as a duodenum, jejunum, or ileum, or the large intestine by utilizing sensors operating with different wavelengths of light. Additionally, ingestible device 100 may be configured to autonomously determine whether it is located within certain portions of the small intestine or large intestine, such as the duodenum, the jejunum, the cecum, or the colon.

[1014] Ingestible device 100 may have a housing 102 shaped similar to a pill or capsule. The housing 102 of ingestible device 100 may have a first end portion 104, and a second end portion 106. The first end portion 104 may include a first wall portion 108, and second end portion 106 may include a second wall portion 110. In some embodiments, first end portion 104 and second end portion 106 of ingestible device 100 may be manufactured separately, and may be affixed together by a connecting portion 112.

[1015] In some embodiments, ingestible device 100 may include an optically transparent window 114. Optically transparent window 114 may be transparent to various types of illumination in the visible spectrum, infrared spectrum, or ultraviolet light spectrum, and ingestible device 100 may have various sensors and illuminators located within the housing 102, and behind the transparent window 114. This may allow ingestible device 100 to be configured to transmit illumination at different wavelengths through transparent window 114 to an environment external to housing 102 of ingestible device 100, and to detect a reflectance from a portion of the illumination that is reflected back through transparent window 114 from the environment external to housing 102. Ingestible device 100 may then use the detected level of reflectance in order to determine a location of ingestible device 100 within a GI tract. In some embodiments, optically transparent window 114 may be of any shape and size, and may wrap around the circumference of ingestible device 100. In this case, ingestible device 100 may have multiple sets of sensors and illuminators positioned at different locations azimuthally behind window 114.

[1016] In some embodiments, ingestible device 100 may optionally include an opening 116 in the second wall portion 110. In some embodiments, the second wall portion 110 may be configured to rotate around the longitudinal axis of ingestible device 100 (e.g., by means of a suitable motor or other actuator housed within ingestible device 100). This may allow ingestible device 100 to obtain a fluid sample from the GI tract, or release a substance into the GI tract, through opening 116.

[1017] FIG. 2 shows an exploded view of ingestible device 100. In some embodiments, ingestible device 100 may optionally include a rotation assembly 118. Optional rotation assembly 118 may include a motor 118-1 driven by a microcontroller (e.g., a microcontroller coupled to printed circuit board 120), a rotation position sensing ring 118-2, and a storage sub-unit 118-3 configured to fit snugly within the second end portion 104. In some embodiments, rotation assembly 118 may cause second end portion 104, and opening 116, to rotate relative to the storage sub-unit 118-3. In some embodiments, there may be cavities on the side of storage sub-unit 118-3 that function as storage chambers. When the opening 116 is aligned with a cavity on the side of the storage sub-unit 118-3, the cavity on the side of the storage sub-unit 118-3 may be exposed to the environment external to the housing 102 of ingestible device 100. In some embodiments, the storage sub-unit 118-3 may be loaded with a medicament or other substance prior to the ingestible device 100 being administered to a subject. In this case, the medicament or other substance may be released from the ingestible device 100 by aligning opening 116 with the cavity within storage sub-unit 118-3. In some embodiments, the storage sub-unit 118-3 may be configured to hold a fluid sample obtained from the GI tract. For example, ingestible device 100 may be configured to align opening 116 with the cavity within storage sub-unit 118-3, thus allowing a fluid sample from the GI tract to enter the cavity within storage sub-unit 118-3. Afterwards, ingestible device 100 may be configured to seal the fluid sample within storage sub-unit 118-3 by further rotating the second end portion 106 relative to storage sub-unit 118-3. In some embodiments, storage sub-unit 118-3 may also contain a hydrophilic sponge, which may enable ingestible device 100 to better draw certain types of fluid samples into ingestible device 100. In some embodiments, ingestible device 100 may be configured to either obtain a sample from within the GI tract, or to release a substance into the GI tract, in response to determining that ingestible device 100 has reached a predetermined location within the GI tract. For example, ingestible device 100 may be configured to obtain a fluid sample from the GI tract in response to determining that the ingestible device has entered the jejunum portion of the small intestine (e.g., as determined by process 900 discussed in relation to FIG. 9). Other ingestible devices capable of obtaining samples or releasing substances are discussed in commonly-assigned PCT Application No. PCT/CA2013/000133 filed Feb. 15, 2013, commonly-assigned U.S. Provisional Application No. 62/385,553, and commonly-assigned U.S. Provisional Application No. 62/376,688, which each are hereby incorporated by reference herein in their entirety. It is understood that any suitable method of obtaining samples or releasing substances may be incorporated into some of the embodiments of the ingestible devices disclosed herein, and that the systems and methods for determining a location of an ingestible device may be incorporated into any suitable type of ingestible device.

[1018] Ingestible device 100 may include a printed circuit board (PCB) 120, and a battery 128 configured to power PCB 120. PCB 120 may include a programmable microcontroller, and control and memory circuitry for holding and executing firmware or software for coordinating the operation of ingestible device 100, and the various components of ingestible device 100. For example, PCB 120 may include memory circuitry for storing data, such as data sets of measurements collected by sensing sub-unit 126, or instructions to be executed by control circuitry to implement a localization process, such as, for example, one or more of the processes, discussed herein, including those discussed below in connection with one or more of the associated flow charts. PCB 120 may include a detector 122 and an illuminator 124, which together form sensing sub-unit 126. In some embodiments, control circuitry within PCB 120 may include processing units, communication circuitry, or any other suitable type of circuitry for operating ingestible device 100. For illustrative purposes, only a single detector 122 and a single illuminator 124 forming a single sensing sub-unit 126 are shown. However, it is understood that in some embodiments there may be multiple sensing sub-units, each with a separate illuminator and detector, within ingestible device 100. For example, there may be several sensing sub-units spaced azimuthally around the circumference of the PCB 120, which may enable ingestible device 100 to transmit illumination and detect reflectances or ambient light in all directions around the circumference of the device. In some embodiments, sensing sub-unit 126 may be configured to generate an illumination using illuminator 124, which is directed through the window 114 in a radial direction away from ingestible device 100. This illumination may reflect off of the environment external to ingestible device 100, and the reflected light coming back into ingestible device 100 through window 114 may be detected as a reflectance by detector 122.

[1019] In some embodiments, window 114 may be of any suitable shape and size. For example, window 114 may extend around a full circumference of ingestible device 100. In some embodiments there may be a plurality of sensing sub-units (e.g., similar to sensing sub-unit 126) located at different positions behind the window. For example, three sensing sub-units may be positioned behind the window at the same longitudinal location, but spaced 120 degrees apart azimuthally. This may enable ingestible device 100 to transmit illuminations in all directions radially around ingestible device 100, and to measure each of the corresponding reflectances.

[1020] In some embodiments, illuminator 124 may be capable of producing illumination at a variety of different wavelengths in the ultraviolet, infrared, or visible spectrum. For example, illuminator 124 may be implemented by using Red-Green-Blue Light-Emitting diode packages (RGB-LED). These types of RGB-LED packages are able to transmit red, blue, or green illumination, or combinations of red, blue, or green illumination. Similarly, detector 122 may be configured to sense reflected light of the same wavelengths as the illumination produced by illuminator 124. For example, if illuminator 124 is configured to produce red, blue, or green illumination, detector 122 may be configured to detect different reflectances produced by red, blue, or green illumination (e.g., through the use of an appropriately configured photodiode). These detected reflectances may be stored by ingestible device 100 (e.g., within memory circuitry of PCB 120), and may then be used by ingestible device 100 in determining a location of ingestible device 100 within the GI tract (e.g., through the use of process 500 (FIG. 5), process 600 (FIG. 6), or process 900 (FIG. 9)).

[1021] It is understood that ingestible device 100 is intended to be illustrative, and not limiting. It will be understood that modifications to the general shape and structure of the various devices and mechanisms described in relation to FIG. 1 and FIG. 2 may be made without significantly changing the functions and operations of the devices and mechanisms. For example, ingestible device 100 may have a housing formed from a single piece of molded plastic, rather than being divided into a first end portion 104 and a second end portion 106. As an alternate example, the location of window 114 within ingestible device 100 may be moved to some other location, such as the center of ingestible device 100, or to one of the ends of ingestible device 100. Moreover, the systems and methods discussed in relation to FIGS. 1-10 may be implemented on any suitable type of ingestible device, provided that the ingestible device is capable of detecting reflectances or levels of illumination in some capacity. For example, in some embodiments ingestible device 100 may be modified to replace detector 122 with an image sensor, and the ingestible device may be configured to measure relative levels of red, blue, or green light by decomposing a recorded image into its individual spectral components. Other examples of ingestible devices with localization capabilities, which may be utilized in order to implement the systems and methods discussed in relation to FIG. 1-11, are discussed in co-owned PCT Application No. PCT/US2015/052500 filed on Sep. 25, 2015, which is hereby incorporated by reference herein in its entirety. Furthermore, it should be noted that the features and limitations described in any one embodiment may be applied to any other embodiment herein, and the descriptions and examples relating to one embodiment may be combined with any other embodiment in a suitable manner.

[1022] FIG. 3 is a diagram of an ingestible device during an example transit through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure. Ingestible device 300 may include any portion of any other ingestible device discussed in this disclosure (e.g., ingestible device 100 (FIG. 1)), and may be any suitable type of ingestible device with localization capabilities. For example, ingestible device 300 may be one embodiment of ingestible device 100 without the optional opening 116 (FIG. 1) or optional rotation assembly 118 (FIG. 2)). In some embodiments, ingestible device 300 may be ingested by a subject, and as ingestible device 300 traverses the GI tract, ingestible device 300 may be configured to determine its location within the GI tract. For example, the movement of ingestible device 300 and the amount of light detected by ingestible device 300 (e.g., via detector 122 (FIG. 2)) may vary substantially depending on the location of ingestible device 300 within the GI tract, and ingestible device 300 may be configured to use this information to determine a location of ingestible device 300 within the GI tract. For instance, ingestible device 300 may detect ambient light from the surrounding environment, or reflectances based on illumination generated by ingestible device 300 (e.g., generated by illuminator 124 (FIG. 1)), and use this information to determine a location of ingestible device 300 through processes, such as described herein. The current location of ingestible device 300, and the time that ingestible device 300 detected each transition between the various portions of the GI tract, may then be stored by ingestible device 300 (e.g., in memory circuitry of PCB 120 (FIG. 2)), and may be used for any suitable purpose.

[1023] Shortly after ingestible device 300 is ingested, ingestible device will traverse the esophagus 302, which may connect the subject's mouth to a stomach 306. In some embodiments, ingestible device 300 may be configured to determine that it has entered the esophagus portion GI tract by measuring the amount and type of light (e.g., via detector 122 (FIG. 2)) in the environment surrounding the ingestible device 300. For instance, ingestible device 300 may detect higher levels of light in the visible spectrum (e.g., via detector 122 (FIG. 2)) while outside the subject's body, as compared to the levels of light detected while within the GI tract. In some embodiments, ingestible device 300 may have previously stored data (e.g., on memory circuitry of PCB 120 (FIG. 2)) indicating a typical level of light detected when outside of the body, and the ingestible device 300 may be configured to determine that entry to the body has occurred when a detected level of light (e.g., detected via detector 122 (FIG. 2)) has been reduced beyond a threshold level (e.g., at least a 20-30% reduction) for a sufficient period of time (e.g., 5.0 seconds).

[1024] In some embodiments, ingestible device 300 may be configured to detect a transition from esophagus 302 to stomach 306 by passing through sphincter 304. In some embodiments, ingestible device 300 may be configured to determine whether it has entered stomach 306 based at least in part on a plurality of parameters, such as but not limited to the use of light or temperature measurements (e.g., via detector 122 (FIG. 2) or via a thermometer within ingestible device 300), pH measurements (e.g., via a pH meter within ingestible device 300), time measurements (e.g., as detected through the use of clock circuitry included within PCB 120 (FIG. 2)), or any other suitable information. For instance, ingestible device 300 may be configured to determine that ingestible device 300 has entered stomach 306 after detecting that a measured temperature of ingestible device 300 exceeds 31 degrees Celsius. Additionally or alternately, ingestible device 300 may be configured to automatically determine it has entered stomach 306 after one minute (or another pre-set time duration parameter, 80 seconds, 90 seconds, etc.) has elapsed from the time that ingestible device 300 was ingested, or one minute (or another pre-set time duration parameter, 80 seconds, 90 seconds, etc.) from the time that ingestible device 300 detected that it has entered the GI tract.

[1025] Stomach 306 is a relatively large, open, and cavernous organ, and therefore ingestible device 300 may have a relatively large range of motion. By comparison, the motion of ingestible device 300 is relatively restricted within the tube-like structure of the duodenum 310, the jejunum 314, and the ileum (not shown), all of which collectively form the small intestine. Additionally, the interior of stomach 306 has distinct optical properties from duodenum 310 and jejunum 314, which may enable ingestible device 300 to detect a transition from stomach 306 to duodenum 310 through the appropriate use of measured reflectances (e.g., through the use of reflectances measured by detector 122 (FIG. 2)), as used in conjunction with process 600 (FIG. 6)).

[1026] In some embodiments, ingestible device 300 may be configured to detect a pyloric transition from stomach 306 to duodenum 310 through the pylorus 308. For instance, in some embodiments, ingestible device 300 may be configured to periodically generate illumination in the green and blue wavelengths (e.g., via illuminator 124 (FIG. 2)), and measure the resulting reflectances (e.g., via detector 122 (FIG. 2)). Ingestible device 300 may be configured to then use a ratio of the detected green reflectance to the detected blue reflectance to determine whether ingestible device 300 is located within the stomach 306, or duodenum 310 (e.g., via process 600 (FIG. 6)). In turn, this may enable ingestible device 300 to detect a pyloric transition from stomach 306 to duodenum 310, an example of which is discussed in relation to FIG. 6.

[1027] Similarly, in some embodiments, ingestible device 300 may be configured to detect a reverse pyloric transition from duodenum 310 to stomach 306. Ingestible device 300 will typically transition naturally from stomach 306 to duodenum 310, and onward to jejunum 314 and the remainder of the GI tract. However, similar to other ingested substances, ingestible device 300 may occasionally transition from duodenum 310 back to stomach 306 as a result of motion of the subject, or due to the natural behavior of the organs with the GI tract. To accommodate this possibility, ingestible device 300 may be configured to continue to periodically generate illumination in the green and blue wavelengths (e.g., via illuminator 124 (FIG. 2)), and measure the resulting reflectances (e.g., via detector 122 (FIG. 2)) to detect whether or not ingestible device 300 has returned to stomach 306. An exemplary detection process is described in additional detail in relation to FIG. 6.

[1028] After entering duodenum 310, ingestible device 300 may be configured to detect a transition to the jejunum 314 through the duodenojejunal flexure 312. For example, ingestible device 300 may be configured to use reflectances to detect peristaltic waves within the jejunum 314, caused by the contraction of the smooth muscle tissue lining the walls of the jejunum 314. In particular, ingestible device 300 may be configured to begin periodically transmitting illumination (and measuring the resulting reflectances (e.g., via detector 122 and illuminator 124 of sensing sub-unit 126 (FIG. 2)) at a sufficiently high frequency in order to detect muscle contractions within the jejunum 314. Ingestible device 300 may then determine that it has entered the jejunum 314 in response to having detected either a first muscle contraction, or a predetermined number of muscle contractions (e.g., after having detected three muscle contractions in sequence). The interaction of ingestible device 300 with the walls of jejunum 314 is also discussed in relation to FIG. 4, and an example of this detection process is described in additional detail in relation to FIG. 9.

[1029] FIG. 4 is a diagram of an ingestible device during an example transit through a jejunum, in accordance with some embodiments of the disclosure. Diagrams 410, 420, 430, and 440 depict ingestible device 400 as it traverses through a jejunum (e.g., jejunum 314), and how ingestible device 400 interacts with peristaltic waves formed by walls 406A and 406B (collectively, walls 406) of the jejunum. In some implementations, ingestible device 400 may include any portion of any other ingestible device discussed in this disclosure (e.g., ingestible device 100 (FIG. 1) or ingestible device 300 (FIG. 3)), and may be any suitable type of ingestible device with localization capabilities. For example, ingestible device 400 may be substantially similar to the ingestible device 300 (FIG. 3) or ingestible device 100 (FIG. 1), with window 404 being the same as window 114 (FIG. 1), and sensing sub-unit 402 being the same as sensing sub-unit 126 (FIG. 2).

[1030] Diagram 410 depicts ingestible device 400 within the jejunum, when the walls 406 of the jejunum are relaxed. In some embodiments, the confined tube-like structure of the jejunum naturally causes ingestible device 400 to be oriented longitudinally along the length of the jejunum, with window 404 facing walls 406. In this orientation, ingestible device 400 may use sensing sub-unit 402 to generate illumination (e.g., via illuminator 124 (FIG. 2)) oriented towards walls 406, and to detect the resulting reflectances (e.g., via detector 122 (FIG. 2)) from the portion of the illumination reflected off of walls 406 and back through window 404. In some embodiments, ingestible device 400 may be configured to use sensing sub-unit 402 to generate illumination and measure the resulting reflectance with sufficient frequency to detect peristaltic waves within the jejunum. For instance, in a healthy human subject, peristaltic waves may occur at a rate of approximately 0.1 Hz to 0.2 Hz. Therefore, the ingestible device 400 may be configured to generate illumination and measure the resulting reflectance at least once every 2.5 seconds (i.e., the minimum rate necessary to detect a 0.2 Hz signal), and preferably at a higher rate, such as once every 0.5 seconds, which may improve the overall reliability of the detection process due to more data points being available. It is understood that the ingestible device 400 need not gather measurements at precise intervals, and in some embodiments the ingestible device 400 may be adapted to analyze data gathered at more irregular intervals, provided that there are still a sufficient number of appropriately spaced data points to detect 0.1 Hz to 0.2 Hz signals.

[1031] Diagram 420 depicts ingestible device 400 within the jejunum, when the walls 406 of the jejunum begin to contract and form a peristaltic wave. Diagram 420 depicts contracting portion 408A of wall 406A and contracting portion 408B of wall 406B (collectively, contracting portion 408 of wall 406) that forma peristaltic wave within the jejunum. The peristaltic wave proceeds along the length of the jejunum as different portions of wall 406 contract and relax, causing it to appear as if contracting portions 408 of wall 406 proceed along the length of the jejunum (i.e., as depicted by contracting portions 408 proceeding from left to right in diagrams 410-430). While in this position, ingestible device 400 may detect a similar level of reflectance (e.g., through the use of illuminator 124 and detector 122 of sensing sub-unit 126 (FIG. 2)) as detected when there is no peristaltic wave occurring (e.g., as detected when ingestible device 400 is in the position indicated in diagram 410).

[1032] Diagram 430 depicts ingestible device 400 within the jejunum, when the walls 406 of the jejunum continue to contract, squeezing around ingestible device 400. As the peristaltic wave proceeds along the length of the jejunum, contracting portions 408 of wall 406 may squeeze tightly around ingestible device 400, bringing the inner surface of wall 406 into contact with window 404. While in this position, ingestible device 400 may detect a change in a reflectance detected as a result of illumination produced by sensing sub-unit 402. The absolute value of the change in the measured reflectance may depend on several factors, such as the optical properties of the window 404, the spectral components of the illumination, and the optical properties of the walls 406. However, ingestible device 400 may be configured to store a data set with the reflectance values over time, and search for periodic changes in the data set consistent with the frequency of the peristaltic waves (e.g., by analyzing the data set in the frequency domain, and searching for peaks between 0.1 Hz to 0.2 Hz). This may enable ingestible device 400 to detect muscle contractions due to peristaltic waves without foreknowledge of the exact changes in reflectance signal amplitude that may occur as a result of detecting the muscle contractions of the peristaltic wave. An example procedure for detecting muscle contractions is discussed further in relation to FIG. 9, and an example of a reflectance data set gathered while ingestible device 400 is located within the jejunum is discussed in relation to FIG. 10.

[1033] Diagram 440 depicts ingestible device 400 within the jejunum, when the peristaltic wave has moved past ingestible device 400. Diagram 440 depicts contracting portions 408 that form the peristaltic wave within the jejunum having moved past the end of ingestible device 400. The peristaltic wave proceeds along the length of the jejunum as different portions of wall 406 contract and relax, causing it to appear as if contracting portions 408 of wall 406 proceed along the length of the jejunum (i.e., as depicted by contracting portions 408 proceeding from left to right in diagrams 410-430). While in this position, ingestible device 400 may detect a similar level of reflectance (e.g., through the use of illuminator 124 and detector 122 of sensing sub-unit 126 (FIG. 2)) as detected when there is no peristaltic wave occurring (e.g., as detected when ingestible device 400 is in the position indicated in diagram 410, or diagram 420).

[1034] Depending on the species of the subject, peristaltic waves may occur relatively with relatively predictable regularity. After the peristaltic wave has passed over ingestible device 400 (e.g., as depicted in diagram 440), the walls 406 of the jejunum may relax again (e.g., as depicted in diagram 410), until the next peristaltic wave begins to form. In some embodiments, ingestible device 400 may be configured to continue to gather reflectance value data while it is within the GI tract, and may store a data set with the reflectance values over time. This may allow ingestible device 400 to detect each of the muscle contractions as the peristaltic wave passes over ingestible device 400 (e.g., as depicted in diagram 430), and may enable ingestible device 400 to both count the number of muscle contractions that occur, and to determine that a current location of the ingestible device 400 is within the jejunum. For example, ingestible device 400 may be configured to monitor for possible muscle contractions while is inside either the stomach or the duodenum, and may determine that ingestible device 400 has moved to the jejunum in response to detecting a muscle contraction consistent with a peristaltic wave.

[1035] FIG. 5 is a flowchart illustrating some aspects of a localization process used by the ingestible device. Although FIG. 5 may be described in connection with the ingestible device 100 for illustrative purposes, this is not intended to be limiting, and either portions or the entirety of the localization procedure 500 described in FIG. 5 may be applied to any device discussed in this application (e.g., the ingestible devices 100, 300, and 400), and any of the ingestible devices may be used to perform one or more parts of the process described in FIG. 5. Furthermore, the features of FIG. 5 may be combined with any other systems, methods or processes described in this application. For example, portions of the process in FIG. 5 may be integrated into or combined with the pyloric transition detection procedure described by FIG. 6, or the jejunum detection process described by FIG. 9.

[1036] At 502, the ingestible device (e.g., ingestible device 100, 300, or 400) gathers measurements (e.g., through detector 122 (FIG. 2)) of ambient light. For example, ingestible device 100 may be configured to periodically measure (e.g., through detector 122 (FIG. 2)) the level of ambient light in the environment surrounding ingestible device 100. In some embodiments, the type of ambient light being measured may depend on the configuration of detector 122 within ingestible device 100. For example, if detector 122 is configured to measure red, green, and blue wavelengths of light, ingestible device 100 may be configured to measure the ambient amount of red, green, and blue light from the surrounding environment. In some embodiments, the amount of ambient light measured by ingestible device 100 will be larger in the area external to the body (e.g., a well-lit room where ingestible device 100 is being administered to a subject) and in the oral cavity of the subject, as compared to the ambient level of light measured by ingestible device 100 when inside of an esophagus, stomach, or other portion of the GI tract (e.g., esophagus 302, stomach 306, duodenum 310, or jejunum 314 (FIG. 3)).

[1037] At 504, the ingestible device (e.g., ingestible device 100, 300, or 400) determines (e.g., via control circuitry within PCB 120 (FIG. 2)) whether the ingestible device has detected entry into the GI tract. For example, ingestible device 100 may be configured to determine when the most recent measurement of ambient light (e.g., the measurement gathered at 502) indicates that the ingestible device has entered the GI tract. For instance, the first time that ingestible device 100 gatherers a measurement of ambient light at 502, ingestible device 100 may store that measurement (e.g., via storage circuitry within PCB 120 (FIG. 2)) as a typical level of ambient light external to the body. Ingestible device 100 may be configured to then compare the most recent measurement of ambient light to the typical level of ambient light external to the body (e.g., via control circuitry within PCB 120 (FIG. 2)), and determine that ingestible device 100 has entered the GI tract when the most recent measurement of ambient light is substantially smaller than the typical level of ambient light external to the body. For example, ingestible device 100 may be configured to detect that it has entered the GI tract in response to determining that the most recent measurement of ambient light is less than or equal to 20% of the typical level of ambient light external to the body. If ingestible device 100 determines that it has detected entry into the GI tract (e.g., that ingestible device 100 has entered at least the esophagus 302 (FIG. 3)), process 500 proceeds to 506. Alternately, if ingestible device 100 determines that it has not detected entry into the GI tract (e.g., as a result of the most recent measurement being similar to the typical level of ambient light external to the body), process 500 proceeds back to 502 where the ingestible device 100 gathers further measurements. For instance, ingestible device 100 may be configured to wait a predetermined amount of time (e.g., five seconds, ten seconds, etc.), and then gather another measurement of the level of ambient light from the environment surrounding ingestible device 100.

[1038] At 506, the ingestible device (e.g., ingestible device 100, 300, or 400) waits for a transition from the esophagus to the stomach (e.g., from esophagus 302 to stomach 306 (FIG. 3)). For example, ingestible device 100 may be configured to determine that it has entered the stomach (e.g., stomach 306 (FIG. 3)) after waiting a predetermined period of time after having entered the GI tract. For instance, a typical esophageal transit time in a human patient may be on the order of 15-30 seconds. In this case, after having detected that ingestible device 100 has entered the GI tract at 504 (i.e., after detecting that ingestible device 100 has reached at least esophagus 302 (FIG. 3)), ingestible device 100 may be configured to wait one minute, or a similar amount of time longer than the typical esophageal transmit time (e.g., ninety-seconds), before automatically determining that ingestible device 100 has entered at least the stomach (e.g., stomach 306 (FIG. 3)).

[1039] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may also determine it has entered the stomach based on measurements of pH or temperature. For example, ingestible device 100 may be configured to determine that it has entered the stomach if a temperature of ingestible device has increased to at least 31 degrees Celsius (i.e., consistent with the temperature inside the stomach), or if a measured pH of the environment surrounding ingestible device 100 is sufficiently acidic (i.e., consistent with the acidic nature of gastric juices that may be found inside the stomach).

[1040] At 508, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating the ingestible device has entered the stomach (e.g., stomach 306 (FIG. 3)). For example, after having waited a sufficient amount of time at 506, ingestible device 100 may store data (e.g., within storage circuitry of PCB 120 (FIG. 2)) indicative of ingestible device 100 having entered at least the stomach. Once ingestible device 100 reaches at least the stomach, process 500 proceeds to 510 where ingestible device 100 may be configured to gather data to detect entry into the duodenum (e.g., duodenum 310 (FIG. 3)).

[1041] In some embodiments, process 500 may also simultaneously proceed from 508 to 520, where ingestible device 100 may be configured to gather data in order to detect muscle contractions and detect entry into the jejunum (e.g., jejunum 314 (FIG. 3)). In some embodiments, ingestible device 100 may be configured to simultaneously monitor for entry into the duodenum at 516-518, as well as detect for entry into the jejunum at 520-524. This may allow ingestible device 100 to determine when it has entered the jejunum (e.g., as a result of detecting muscle contractions), even when it fails to first detect entry into the duodenum (e.g., as a result of very quick transit times of the ingestible device through the duodenum).

[1042] At 510, the ingestible device (e.g., ingestible device 100, 300, or 400) gathers measurements of green and blue reflectance levels (e.g., through the use of illuminator 124 and detector 122 of sensing sub-unit 126 (FIG. 2)) while in the stomach (e.g., stomach 306 (FIG. 3)). For example, ingestible device 100 may be configured to periodically gather measurements of green and blue reflectance levels while in the stomach. For instance, ingestible device 100 may be configured to transmit a green illumination and a blue illumination (e.g., via illuminator 124 (FIG. 2)) every five to fifteen seconds, and measure the resulting reflectance (e.g., via detector 122 (FIG. 2)). Every time that ingestible device 100 gathers a new set of measurements, the measurements may be added to a stored data set (e.g., stored within memory circuitry of PCB 120 (FIG. 2)). The ingestible device 100 may then use this data set to determine whether or not ingestible device 100 is still within a stomach (e.g., stomach 306 (FIG. 3)), or a duodenum (e.g., duodenum 310 (FIG. 3)).

[1043] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may be configured to detect a first reflectance based on generating an illumination of a first wavelength in approximately the green spectrum of light (between 495-600 nm), and detecting a second reflectance based on generating an illumination of the second wavelength in approximately the blue spectrum of light (between 400-495 nm). In some embodiments, the ingestible device may ensure that the illumination in the green spectrum and the illumination in the blue spectrum have wavelengths separated by at least 50 nm. This may enable ingestible device 100 to sufficiently distinguish between the two wavelengths when detecting the reflectances (e.g., via detector 122 (FIG. 2)). It is understood that the separation of 50 nm is intended to be illustrative, and not limiting, and depending on the accuracy of the detectors within ingestible device 100, smaller separations may be possible to be used.

[1044] At 512, the ingestible device (e.g., ingestible device 100, 300, or 400) determines (e.g., using control circuitry within PCB 120 (FIG. 2)) whether the ingestible device has detected a transition from the stomach (e.g., stomach 306 (FIG. 3)) to a duodenum (e.g., duodenum 310 (FIG. 3)) based on a ratio of green and blue (G/B) reflectance levels. For example, ingestible device 100 may obtain (e.g., from memory circuitry of PCB 120 (FIG. 2)) a data set containing historical data for the respective ratio of the green reflectance to the blue reflectance as measured at a respective time. Generally speaking, a duodenum (e.g., duodenum 310 (FIG. 3)) of a human subject reflects a higher ratio of green light to blue light, as compared to the ratio of green light to blue light that is reflected by a stomach (e.g., stomach 306 (FIG. 3)). Based on this, ingestible device 100 may be configured to take a first set of ratios from the data set, representing the result of recent measurements, and compare them to a second set of ratios from the data set, representing the results of past measurements. When the ingestible device 100 determines that the mean value of the first set of ratios is substantially larger than the mean value of the second set of ratios (i.e., that the ratio of reflected green light to reflected blue light has increased), the ingestible device 100 may determine that it has entered the duodenum (e.g., duodenum 310 (FIG. 3)) from the stomach (e.g., stomach 306 (FIG. 3)). If the ingestible device 100 detects a transition from the stomach (e.g., stomach 306 (FIG. 3)) to a duodenum (e.g., duodenum 310 (FIG. 3)), process 500 proceeds to 514, where ingestible device 100 stores data indicating that the ingestible device 100 has entered the duodenum (e.g., duodenum 310 (FIG. 3)). Alternatively, if the ingestible device determines that the ingestible device has not transitioned from the stomach (e.g., stomach 306 (FIG. 3)) to the duodenum (e.g., duodenum 310 (FIG. 3)), process 500 proceeds back to 510 to gather more measurements of green and blue reflectance levels while still in the stomach (e.g., stomach 306 (FIG. 3)). An example procedure for using measurements of green and blue reflectances to monitor for transitions between the stomach and the duodenum is discussed in greater detail in relation to FIG. 6.

[1045] In some embodiments, the first time that ingestible device 100 detects a transition from the stomach (e.g., stomach 306 (FIG. 3)) to the duodenum (e.g., duodenum 310 (FIG. 3)), ingestible device 100 may be configured to take a mean of the second set of data, (e.g., the set of data previously recorded while in stomach 306 (FIG. 3)) and store this as a typical ratio of green light to blue light detected within the stomach (e.g., stomach 306 (FIG. 3)) (e.g., within memory circuitry of PCB 120 (FIG. 2)). This stored information may later be used by ingestible device 100 to determine when ingestible device 100 re-enters the stomach (e.g., stomach 306 (FIG. 3)) from the duodenum (e.g., duodenum 310 (FIG. 3)) as a result of a reverse pyloric transition.

[1046] At 514, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating that the ingestible device has entered the duodenum (e.g., duodenum 310 (FIG. 3)). For example, ingestible device 100 may store a flag within local memory (e.g., memory circuitry of PCB 120) indicating that the ingestible device 100 is currently in the duodenum. In some embodiments, the ingestible device 100 may also store a timestamp indicating the time when ingestible device 100 entered the duodenum. Once ingestible device 100 reaches the duodenum, process 500 proceeds to 520 where ingestible device 100 may be configured to gather data in order to detect muscle contractions and detect entry into the jejunum (e.g., jejunum 314 (FIG. 3)). Process 500 also proceeds from 514 to 516, where ingestible device 100 may be configured to gather data additional data in order to detect re-entry into the stomach (e.g., stomach 306 (FIG. 3)) from the duodenum (e.g., duodenum 310 (FIG. 3)).

[1047] At 516, the ingestible device (e.g., ingestible device 100, 300, or 400) gathers measurements (e.g., via sensing sub-unit 126 (FIG. 2)) of green and blue reflectance levels while in the duodenum (e.g., duodenum 310 (FIG. 3)). For example, ingestible device 100 may be configured to periodically gather measurements (e.g., via sensing sub-unit 126 (FIG. 2)) of green and blue reflectance levels while in the duodenum, similar to the measurements made at 510 while in the stomach. For instance, ingestible device 100 may be configured to transmit a green illumination and a blue illumination (e.g., via illuminator 124 (FIG. 2)) every five to fifteen seconds, and measure the resulting reflectance (e.g., via detector 122 (FIG. 2)). Every time that ingestible device 100 gathers a new set of measurements, the measurements may be added to a stored data set (e.g., stored within memory circuitry of PCB 120 (FIG. 2)). The ingestible device 100 may then use this data set to determine whether or not ingestible device 100 is still within the duodenum (e.g., duodenum 310 (FIG. 3)), or if the ingestible device 100 has transitioned back into the stomach (e.g., stomach 306 (FIG. 3)).

[1048] At 518, the ingestible device (e.g., ingestible device 100, 300, or 400) determines a transition from the duodenum (e.g., duodenum 310 (FIG. 3)) to the stomach (e.g., stomach 306 (FIG. 3)) based on a ratio of the measured green reflectance levels to the measured blue reflectance levels. In some embodiments, ingestible device 100 may compare the ratio of the measured green reflectance levels to the measured blue reflectance levels recently gathered by ingestible device 100 (e.g., measurements gathered at 516), and determine whether or not the ratio of the measured green reflectance levels to the measured blue reflectance levels is similar to the average ratio of the measured green reflectance levels to the measured blue reflectance levels seen in the stomach (e.g., stomach 306 (FIG. 3)). For instance, ingestible device 100 may retrieve data (e.g., from memory circuitry of PCB 120 (FIG. 2)) indicative of the average ratio of the measured green reflectance levels to the measured blue reflectance levels seen in the stomach, and determine that ingestible device 100 has transitioned back to the stomach if the recently measured ratio of the measured green reflectance levels to the measured blue reflectance levels is sufficiently similar to the average level in the stomach (e.g., within 20% of the average ratio of the measured green reflectance levels to the measured blue reflectance levels seen in the stomach, or within any other suitable threshold level). If the ingestible device detects a transition from the duodenum (e.g., duodenum 310 (FIG. 3)) to the stomach (e.g., stomach 306 (FIG. 3)), process 500 proceeds to 508 to store data indicating the ingestible device has entered the stomach (e.g., stomach 306 (FIG. 3)), and continues to monitor for further transitions. Alternatively, if the ingestible device does not detect a transition from the duodenum (e.g., duodenum 310 (FIG. 3)) to the stomach (e.g., stomach 306 (FIG. 3)), process 500 proceeds to 516 to gather additional measurements of green and blue reflectance levels while in the duodenum (e.g., duodenum 310 (FIG. 3)), which may be used to continuously monitor for possible transitions back into the stomach. An example procedure for using measurements of green and blue reflectances to monitor for transitions between the stomach and the duodenum is discussed in greater detail in relation to FIG. 6.

[1049] At 520, the ingestible device (e.g., ingestible device 100, 300, or 400) gathers periodic measurements of the reflectance levels (e.g., via sensing sub-unit 126 (FIG. 2)) while in the duodenum (e.g., duodenum 310 (FIG. 3)). In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may gather similar periodic measurements while in the stomach as well. In some embodiments, these periodic measurements may enable ingestible device 100 to detect muscle contractions (e.g., muscle contractions due to a peristaltic wave as discussed in relation to FIG. 4), which may be indicative of entry into a jejunum (e.g., jejunum 314 (FIG. 3)). Ingestible device 100 may be configured to gather periodic measurements using any suitable wavelength of illumination (e.g., by generating illumination using illuminator 124, and detecting the resulting reflectance using detector 122 (FIG. 2)), or combinations of wavelengths of illumination. For example, in some embodiments, ingestible device 100 may be configured to generate red, green, and blue illumination, store separate data sets indicative of red, green, and blue illumination, and analyze each of the data sets separately to search for frequency components in the recorded data indicative of detected muscle contractions. In some embodiments, the measurements gathered by ingestible device 100 at 520 may be sufficiently fast as to detect peristaltic waves in a subject. For instance, in a healthy human subject, peristaltic waves may occur at a rate of approximately 0.1 Hz to 0.2 Hz. Therefore, the ingestible device 400 may be configured to generate illumination and measure the resulting reflectance at least once every 2.5 seconds (i.e., the minimum rate necessary to detect a 0.2 Hz signal), and preferably at a higher rate, such as once every 0.5 seconds or faster, and store values indicative of the resulting reflectances in a data set (e.g., within memory circuitry of PCB 120 (FIG. 2)). After gathering additional data (e.g., after gathering one new data point, or a predetermined number of new data points), process 500 proceeds to 522, where ingestible device 100 determines whether or not a muscle contraction has been detected.

[1050] At 522, the ingestible device (e.g., ingestible device 100, 300, or 400) determines (e.g., via control circuitry within PCB 120 (FIG. 0.2)) whether the ingestible device detects a muscle contraction based on the measurements of reflectance levels (e.g., as gathered by sensing sub-unit 126 (FIG. 2)). For example, ingestible device 100 may obtain a fixed amount of data stored as a result of measurements made at 520 (e.g., retrieve the past minute of data from memory circuitry within PCB 120 (FIG. 2)). Ingestible device 100 may then convert the obtained data into the frequency domain, and search for peaks in a frequency range that would be consistent with peristaltic waves. For example, in a healthy human subject, peristaltic waves may occur at a rate of approximately 0.1 Hz to 0.2 Hz, and an ingestible device 100 may be configured to search for peaks in the frequency domain representation of the data between 0.1 Hz and 0.2 Hz above a threshold value. If the ingestible device 100 detects a contraction based on the reflectance levels (e.g., based on detecting peaks in the frequency domain representation of the data between 0.1 Hz and 0.2 Hz), process 500 proceeds to 524 to store data indicating that the device has entered the jejunum. Alternatively, if the ingestible device 100 does not detect a muscle contraction, process 500 proceeds to 520 to gather periodic measurements of the reflectance levels while in the duodenum (e.g., duodenum 310 (FIG. 3)). In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may store data (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating that a muscle contraction was detected, and process 500 will not proceed from 522 to 524 until a sufficient number of muscle contractions have been detected.

[1051] At 524, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating that the device has entered the jejunum (e.g., jejunum 314 (FIG. 3)). For example, in response to detecting that muscle contraction has occurred at 522, ingestible device 100 may determine that it has entered the jejunum 314, and is no longer inside of the duodenum (e.g., duodenum 310 (FIG. 3)) or the stomach (e.g., stomach 306 (FIG. 3)). In some embodiments, the ingestible device 100 may continue to measure muscle contractions while in the jejunum, and may store data indicative of the frequency, number, or strength of the muscle contractions over time (e.g., within memory circuitry of PCB 120 (FIG. 2)). In some embodiments, the ingestible device 100 may also be configured to monitor for one or more transitions. Such transitions can include a transition from the jejunum to the ileum, an ileoceacal transition from the ileum to the cecum, a transition from the cecum to the colon, or detect exit from the body (e.g., by measuring reflectances, temperature, or levels of ambient light).

[1052] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may also determine that it has entered the jejunum (e.g., jejunum 314 (FIG. 3)) after a pre-determined amount of time has passed after having detected entry into the duodenum (e.g., duodenum 310 (FIG. 3)). For example, barring a reverse pyloric transition from the duodenum (e.g., duodenum 310 (FIG. 3)) back to the stomach (e.g., stomach 306 (FIG. 3)), the typical transit time for an ingestible device to reach the jejunum from the duodenum in a healthy human subject is less than three minutes. In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may therefore be configured to automatically determine that it has entered the jejunum after spending at least three minutes within the duodenum. This determination may be made separately from the determination made based on measured muscle contractions (e.g., the determination made at 522), and in some embodiments, ingestible device 100 may determine that it has entered the jejunum in response to either detecting muscle contractions, or after three minutes has elapsed from having entered the duodenum (e.g., as determined by storing data at 514 indicative of the time that ingestible device entered the duodenum).

[1053] For illustrative purposes, 512-518 of process 500 describe the ingestible device (e.g., ingestible device 100, 300, or 400) measuring green reflectances and blue reflectances, calculating a ratio of the two reflectances, and using this information to determine when the ingestible device has transitioned between the duodenum and stomach. However, in some embodiments, other wavelengths of light may be used other than green and blue, provided that the wavelengths of light chosen have different reflective properties within the stomach and the duodenum (e.g., as a result of different reflection coefficients of the stomach tissue and the tissue of the duodenum).

[1054] It will be understood that the steps and descriptions of the flowcharts of this disclosure, including FIG. 5, are merely illustrative. Any of the steps and descriptions of the flowcharts, including FIG. 5, may be modified, omitted, rearranged, and performed in alternate orders or in parallel, two or more of the steps may be combined, or any additional steps may be added, without departing from the scope of the present disclosure. For example, the ingestible device 100 may calculate the mean and the standard deviation of multiple data sets in parallel in order to speed up the overall computation time. As another example, ingestible device 100 may gather data periodic measurements and detect possible muscle contractions (e.g., at 520-522) while simultaneously gathering green and blue reflectance levels to determine transitions to and from the stomach and duodenum (e.g., at 510-518). Furthermore, it should be noted that the steps and descriptions of FIG. 5 may be combined with any other system, device, or method described in this application, including processes 600 (FIG. 6) and 900 (FIG. 9), and any of the ingestible devices or systems discussed in this application (e.g., ingestible devices 100, 300, or 400) could be used to perform one or more of the steps in FIG. 5.

[1055] FIG. 6 is a flowchart illustrating some aspects of a process for detecting transitions from a stomach to a duodenum and from a duodenum back to a stomach, which may be used when determining a location of an ingestible device as it transits through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure. In some embodiments, process 600 may begin when an ingestible device first detects that it has entered the stomach, and will continue as long as the ingestible device determines that it is within the stomach or the duodenum. In some embodiments, process 600 may only be terminated when an ingestible device determines that it has entered the jejunum, or otherwise progressed past the duodenum and the stomach. Although FIG. 6 may be described in connection with the ingestible device 100 for illustrative purposes, this is not intended to be limiting, and either portions or the entirety of the duodenum detection process 600 described in FIG. 6 may be applied to any device discussed in this application (e.g., the ingestible devices 100, 300, or 400), and any of the ingestible devices may be used to perform one or more parts of the process described in FIG. 6. Furthermore, the features of FIG. 6 may be combined with any other systems, methods or processes described in this application. For example, portions of the process described by the process in FIG. 6 may be integrated into process 500 discussed in relation to FIG. 5.

[1056] At 602, the ingestible device (e.g., ingestible device 100, 300, or 400) retrieves a data set (e.g., from memory circuitry within PCB 120 (FIG. 2)) with ratios of the measured green reflectance levels to the measured blue reflectance levels over time. For example, ingestible device 100 may retrieve a data set from PCB 120 containing recently recorded ratios of the measured green reflectance levels to the measured blue reflectance levels (e.g., as recorded at 510 or 516 of process 500 (FIG. 5)). In some embodiments, the retrieved data set may include the ratios of the measured green reflectance levels to the measured blue reflectance levels over time. Example plots of data sets of ratios of the measured green reflectance levels to the measured blue reflectance levels are discussed further in relation to FIG. 7 and FIG. 8.

[1057] At 604, the ingestible device (e.g., ingestible device 100, 300, or 400) includes a new measurement (e.g., as made with sensing sub-unit 126 (FIG. 2)) of a ratio of the measured green reflectance level to the measured blue reflectance level in the data set. For example, ingestible device 100 may be configured to occasionally record new data by transmitting green and blue illumination (e.g., via illuminator 124 (FIG. 2)), detecting the amount of reflectance received due to the green and blue illumination (e.g., via detector 122 (FIG. 2)), and storing data indicative of the amount of the received reflectance (e.g., in memory circuitry of PCB 120 (FIG. 2)). The ingestible device 100 may be configured to record new data every five to fifteen seconds, or at any other convenient interval of time. For illustrative purposes, ingestible device 100 is described as storing and retrieving the ratio of the measured green reflectance levels to the measured blue reflectance levels (e.g., if the amount of detected green reflectance was identical to the amount of detected blue reflectance at a given time, the ratio of the green and blue reflectances would be 1.0 at that given time); however, it is understood that the green reflectance data and the blue reflectance data may be stored separately within the memory of ingestible device 100 (e.g., stored as two separate data sets within memory circuitry of PCB 120 (FIG. 2)).

[1058] At 606, the ingestible device (e.g., ingestible device 100, 300, or 400) retrieves a first subset of recent data by applying a first sliding window filter to the data set. For example, ingestible device 100 may use a sliding window filter to obtain a predetermined amount of the most recent data within the data set, which may include any new values of the ratio of the measured green reflectance level to the measured blue reflectance level obtained at 604. For instance, the ingestible device may be configured to select between ten and forty data points from the data set, or ingestible device 100 may be configured to select a predetermined range of data values between fifteen seconds of data and five minutes of data. In some embodiments, other ranges of data may be selected, depending on how frequently measurements are recorded, and the particular application at hand. For instance, any suitable amount of data may be selected in the sliding window, provided that it is sufficient to detect statistically significant differences between the data selected in a second sliding window (e.g., the second subset of data selected at 614).

[1059] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may also be configured to remove outliers from the data set, or to smooth out unwanted noise in the data set. For example, ingestible device 100 may select the first subset of data, or any other subset of data, by first obtaining a raw set of values by applying a window filter to the data set (e.g., selecting a particular range of data to be included). Ingestible device 100 may then be configured to identify outliers in the raw set of values; for instance, by identifying data points that are over three standard deviations away from the mean value of the raw set of values, or any other suitable threshold. Ingestible device 100 may then determine the subset of data by removing outliers from the raw set of values. This may enable ingestible device 100 to avoid spurious information when determining whether or not it is located within the stomach or the duodenum.

[1060] At 608, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether the most recently detected location was the duodenum (e.g., duodenum 310 (FIG. 3)). In some embodiments, ingestible device 100 may store a data flag (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating the most recent portion of the GI tract that the ingestible device 100 detected itself to be within. For instance, every time ingestible device 100 detects entry to the stomach (e.g., detects entry into stomach 306 (FIG. 3) as a result of the decision made at 610), a flag is stored in memory indicating the ingestible device 100 is in the stomach (e.g., as part of storing data at 612). If ingestible device 100 subsequently detects entry into the duodenum (e.g., detects entry into duodenum 310 (FIG. 3) as a result of a decision made at 624), another different flag is stored in memory indicating that the ingestible device 100 is in the duodenum (e.g., as part of storing data at 624). In this case, ingestible device 100 may retrieve the most recently stored flag at 608, and determine whether or not the flag indicates that the ingestible device 100 was most recently within the duodenum. If ingestible device 100 detects that it was most recently in the duodenum, process 600 proceeds to 610 where the ingestible device compares the recent measurements of the ratios of the measured green reflectance levels to the measured blue reflectance levels (e.g., measurements that include the recent measurement made at 606) to the typical ratios measured within the stomach, and uses this information to determine whether a reverse pyloric transition from the duodenum back to the stomach has occurred. Alternately, if ingestible device 100 detects that it was not most recently in the duodenum (e.g., because it was in the stomach instead), process 600 proceeds to 614 where the ingestible device compares the recent measurements of the ratios of the measured green reflectance levels to the measured blue reflectance levels (e.g., measurements that include the recent measurement made at 606) to past measurements, and uses this information to determine whether a pyloric transition from the stomach to the duodenum has occurred.

[1061] Process 600 proceeds from 608 to 610 when the ingestible device determined that it was most recently in the duodenum. At 610, the ingestible device (e.g., ingestible device 100, 300, or 400) determines (e.g., via control circuitry within PCB 120 (FIG. 2)) whether the current G/B signal is similar to a recorded average G/B signal in the stomach. For example, ingestible device 100 may be configured to have previously stored data (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicative of the average ratio of the measured green reflectance levels to the measured blue reflectance levels measured in the stomach. Ingestible device 100 may then retrieve this stored data indicative of the average ratio of the measured green reflectance levels to the measured blue reflectance levels in the stomach, and compare this against the recent measurements in order to determine whether or not ingestible device 100 has returned back to the stomach from the duodenum. For instance, ingestible device 100 may determine if the mean value of the first subset of recent data (i.e., the average value of the recently measured ratios of the measured green reflectance levels to the measured blue reflectance levels) is less than the average ratio of the measured green reflectance levels to the measured blue reflectance levels within the stomach, or less that the average ratio measured within the stomach plus a predetermined number times the standard deviation of the ratios measured within the stomach. For instance, if the average ratio of the measured green reflectance levels to the measured blue reflectance levels in the stomach was 1, with a standard deviation of 0.2, ingestible device 100 may determine whether or not the mean value of the first subset of data is less than 1.0+k*0.2, where k is a number between zero and five. It is understood that, in some embodiments, the ingestible device 100 may be configured to use a different threshold level to determine whether or not the mean value of the first subset of recent data is sufficiently similar to the average ratio of the measured green reflectance levels to the measured blue reflectance levels within the stomach. In response to determining that the recent ratio of the measured green reflectance levels to the measured blue reflectance levels is similar to the average ratio of measured green and blue reflectance levels seen in the stomach, process 600 proceeds to 612 where ingestible device 100 stores data indicating that it has re-entered the stomach from the duodenum. Alternately, in response to determining that the recent ratio of measured green and blue reflectance levels is sufficiently different from the average ratio of measured green and blue reflectance levels seen in the stomach, ingestible device 100 proceeds directly to 604, and continues to obtain new data on an ongoing basis.

[1062] At 612, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating a reverse pyloric transition from the duodenum to the stomach was detected. For example ingestible device 100 may store a data flag (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating that the ingestible device 100 most recently detected itself to be within the stomach portion of the GI tract (e.g., stomach 306 (FIG. 3)). In some embodiments, ingestible device 100 may also store data (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating a time that ingestible device 100 detected the reverse pyloric transition from the duodenum to the stomach. This information may be used by ingestible device 100 at 608, and as a result process 600 may proceed from 608 to 614, rather than proceeding from 618 to 610. After ingestible device 100 stores the data indicating a reverse pyloric transition from the duodenum to the stomach was detected, process 600 proceeds to 604 where ingestible device 100 continues to gather additional measurements, and continues to monitor for further transitions between the stomach and the duodenum.

[1063] Process 600 proceeds from 608 to 614 when the ingestible device determined that it was not most recently in the duodenum (e.g., as a result of having most recently been in the stomach instead). At 614, the ingestible device (e.g., ingestible device 100, 300, or 400) retrieves a second subset of previous data by applying a second sliding window filter to the data set. For example, ingestible device 100 may use a sliding window filter to obtain a predetermined amount of older data from a past time range, which may be separated from recent time range used to select the first subset of data gathered at 606 by a predetermined period of time. In some embodiments, any suitable amount of data may be selected by the first and second window filters, and the first and second window filters may be separated by any appropriate predetermined amount of time. For example, in some embodiments, the first window filter and the second window filter may each be configured to select a predetermined range of data values from the data set, the predetermined range being between fifteen seconds of data and five minutes of data. In some embodiments, the recent measurements and the past measurements may then be separated by a predetermined period of time that is between one to five times the predetermined range of data values. For instance, ingestible device 100 may select the first subset of data and the second subset of data to each be one minute of data selected from the dataset (i.e., selected to have a predetermined range of one minute), and the first subset of data and the second subset of data are selected from recorded measurements that are at least two minutes apart (i.e., the predetermined period of time is two minutes, which is twice the range used to select the subsets of data using the window filters). As another example, ingestible device 100 may select the first subset of data and the second subset of data to each be five minutes of data selected from the dataset (i.e., selected to have a predetermined range of five minutes), and the first subset of data and the second subset of data are selected from recorded measurements that are at least 10 minutes apart (i.e., the predetermined period of time is two minutes, which is twice the range used to select the subsets of data using the window filters).

[1064] In some embodiments, if ingestible device 100 recently transitioned to the stomach from the duodenum (e.g., as determined by checking for recent data stored within ingestible device 100 at 612), ingestible device 100 may select the second subset of data at 614 from a time frame when ingestible device 100 is known to be within the stomach. In some embodiments, ingestible device 100 may alternately select a previously recorded average and standard deviation for ratios of green reflectances and blue reflectances within the stomach (e.g., an average and standard deviation typical of data recorded within the stomach, as previously recorded within memory circuitry of PCB 120 at 620) in place of the second subset of data. In this case, ingestible device 100 may simply use the previously recorded average and previously recorded standard deviation when making a determination at 616, rather than expending resources to calculate the mean and standard deviation of the second subset.

[1065] At 616, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether the difference between the mean of the second subset and the mean of the first subset is greater than a predetermined multiple of the standard deviation of the first subset. For example, ingestible device 100 may compute a difference between a mean of the first subset of recent data and a mean of a second subset of past data, and determine whether this difference is greater than three times the standard deviation of the second subset of past data. In some embodiments, it is understood that any convenient threshold level may be used other than three times the standard deviation, such as any value between one and five times the standard deviation. Also, in some embodiments, the ingestible device may instead set the threshold level based on the standard deviation of the second subset instead of the first subset. In response to determining that the difference between the mean of the first subset and the mean of the second subset is greater than a predetermined multiple of the standard deviation of the second subset, process 600 proceeds to 618. Otherwise, process 600 proceeds back to 604, where the ingestible device 604 continues to gather new data to be used in monitoring for transitions between the stomach (e.g., stomach 306 (FIG. 3)) and the duodenum (e.g., duodenum 310 (FIG. 3)).

[1066] At 618, the ingestible device (e.g., ingestible device 100, 300, or 400) determines (e.g., via control circuitry within PCB 120 (FIG. 2)) whether the determination made at 616 is the first time that the difference between the mean of the first subset of recent data and the mean of the second subset of past data is calculated to be greater than the standard deviation of the second subset. If the ingestible device determines that this is the first time that the difference between the mean of the first subset and the mean of the second subset is calculated to be greater than the standard deviation of the second subset, process 600 proceeds to 620 to store the mean of the second subset of past data as an average G/B signal in the stomach. Alternatively, if the ingestible device determines that the immediately preceding determination made at 616 is not the first time that the difference between the mean of the first subset of recent data and the mean of the second subset of past data is calculated to be greater than the standard deviation of the second subset, process 600 proceeds directly to 622.

[1067] At 620, the ingestible device (e.g., ingestible device 100, 300, or 400) stores the mean of the second subset as an average G/B signal in the stomach. For example, ingestible device 100 may be configured to store the mean of the second subset of past data (e.g., store within memory circuitry of PCB 120 (FIG. 2)) as the average ratio of the measured green reflectance levels to the measured blue reflectance levels measured in the stomach. In some embodiments, ingestible device 100 may also store the standard deviation of the second subset of past data as a typical standard deviation of the ratios of the measured green reflectance levels to the measured blue reflectance levels detected within the stomach. This stored information may be used by the ingestible device later on (e.g., at 610) to compare against future data, which may enable the ingestible device to detect reverse pyloric transitions from the duodenum (e.g., duodenum 310 (FIG. 3)) back to the stomach (e.g., stomach 306 (FIG. 3)), and may generally be used in place of other experimental data gathered from the stomach (e.g., in place of the second subset of data at 616). After storing the mean of the second subset as an average G/B signal in the stomach, process 600 proceeds to 622.

[1068] At 622, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether a difference of the mean of the first subset of recent data to the mean of the second subset of past data is greater than a predetermined threshold, M. In some embodiments, the predetermined threshold, M, will be sufficiently large to ensure that the mean of the first subset is substantially larger than the mean of the second subset, and may enable ingestible device 100 to ensure that it detected an actual transition to the duodenum. This may be particularly advantageous when the determination made at 616 is potentially unreliable due to the standard deviation of the second subset of past data being abnormally small. For example, a typical value of the predetermined threshold M, may be on the order of 0.1 to 0.5. If ingestible device 100 determines that the difference of the mean of the first subset of recent data to the second subset of past data is greater than a predetermined threshold, process 600 proceeds to 624 to store data indicating that a pyloric transition from the stomach to the duodenum (e.g., from stomach 306 to duodenum 310 (FIG. 3)) was detected. Alternatively, if the ingestible device determines that the ratio of the mean of the first subset to the second subset is less than or equal to the predetermined threshold, M (i.e., determines that a transition to the duodenum has not occurred), process 600 proceeds directly to 604 where ingestible device 100 continues to make new measurements and monitor for possible transitions between the stomach and the duodenum.

[1069] In some embodiments, instead of using a difference of the mean of the first subset of recent data to the mean of the second subset of past data, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether the ratio of the mean of the first subset of recent data to the mean of the second subset of past data is greater than a predetermined threshold, M. In some embodiments, the predetermined threshold, M, will be sufficiently large to ensure that the mean of the first subset is substantially larger than the mean of the second subset, and may enable ingestible device 100 to ensure that it detected an actual transition to the duodenum. This may be particularly advantageous when the determination made at 616 is potentially unreliable due to the standard deviation of the second subset of past data being abnormally small. For example, a typical value of the predetermined threshold M, may be on the order of 1.2 to 2.0. It is understood any convenient type of threshold or calculation may be used to determine whether or not the first subset of data and the second subset of data are both statistically distinct from one another, and also substantially different from one another in terms of overall average value.

[1070] At 624, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating a pyloric transition from the stomach to the duodenum was detected. For example ingestible device 100 may store a data flag (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating that the ingestible device 100 most recently detected itself to be within the duodenum portion of the GI tract (e.g., duodenum 310 (FIG. 3)). In some embodiments, ingestible device 100 may also store data (e.g., within memory circuitry of PCB 120 (FIG. 2)) indicating a time that ingestible device 100 detected the pyloric transition from the stomach to the duodenum. This information may be used by ingestible device 100 at 608, and as a result process 600 may proceed from 608 to 610, rather than proceeding from 618 to 614. After ingestible device 100 stores the data indicating a pyloric transition from the stomach to the duodenum was detected, process 600 proceeds to 604 where ingestible device 100 continues to gather additional measurements, and continues to monitor for further transitions between the stomach and the duodenum.

[1071] It will be understood that the steps and descriptions of the flowcharts of this disclosure, including FIG. 6, are merely illustrative. Any of the steps and descriptions of the flowcharts, including FIG. 6, may be modified, omitted, rearranged, and performed in alternate orders or in parallel, two or more of the steps may be combined, or any additional steps may be added, without departing from the scope of the present disclosure. For example, the ingestible device 100 may calculate the mean and the standard deviation of multiple data sets in parallel in order to speed up the overall computation time. Furthermore, it should be noted that the steps and descriptions of FIG. 6 may be combined with any other system, device, or method described in this application, and any of the ingestible devices or systems discussed in this application could be used to perform one or more of the steps in FIG. 6. For example, portions of process 600 may be incorporated into 508-516 of process 500 (FIG. 5), and may be part of a more general process for determining a location of the ingestible device. As another example, the ratio of detected blue and green light (e.g., as measured and added to the data set at 604) may continue even outside of the stomach or duodenum, and similar information may be recorded by the ingestible device throughout its transit in the GI tract. Example plots of data sets of ratios of measured green and blue reflectance levels, which may be gathered throughout the GI tract, are discussed further in relation to FIG. 7 and FIG. 8 below.

[1072] FIG. 7 is a plot illustrating data collected during an example operation of an ingestible device (e.g., ingestible device 100, 300, or 400), which may be used when determining a location of an ingestible device as it transits through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure.

Although FIG. 7 may be described in connection with ingestible device 100 for illustrative purposes, this is not intended to be limiting, and plot 700 and data set 702 may be typical of data gathered by any device discussed in this application. Plot 700 depicts the ratios of the measured green reflectance levels to the measured blue reflectance levels over time. For example, ingestible device 100 may have computed the value for each point in the data set 702 by transmitting green and blue illumination at a given time (e.g., via illuminator 124 (FIG. 2)), measuring the resulting green and blue reflectances (e.g., via detector 122 (FIG. 2)), calculating the ratio of the resulting reflectances, and storing the ratio in the data set along with a timestamp indicating the time that the reflectances were gathered.

[1073] At 704, shortly after ingestible device 100 begins operation, ingestible device 100 determines that it has reached at least the stomach (e.g., as a result of making a determination similar to the determination discussed in relation to 506 in process 500 (FIG. 5)). Ingestible device 100 continues to gather additional measurements of green and blue reflectance levels, and at 706 ingestible device 100 determines that a pyloric transition has occurred from the stomach to the duodenum (e.g., as a result of making a determination similar to the determinations discussed in relation to 616-624 of process 600 (FIG. 6)). Notably, the values in data set 702 around 706 jump up precipitously, which is indicative of the higher ratios of measured green reflectance levels to measured blue reflectance levels typical of the duodenum.

[1074] The remainder of the data set 702 depicts the ratios of the measured green reflectance levels to the measured blue reflectance levels throughout the remainder of the GI tract. At 708, ingestible device 100 has reached the jejunum (e.g., as determined through measurements of muscle contractions, as discussed in relation to FIG. 9), and by 710, ingestible device 100 has reached the cecum. It is understood that, in some embodiments, the overall character and appearance of data set 702 changes within the small intestine (i.e., the duodenum, jejunum, and ileum) versus the cecum. Within the jejunum and ileum, there may typically be a wide variation in the ratios of the measured green reflectance levels to the measured blue reflectance levels, resulting in relatively noisy data with a high standard deviation. By comparison, within the cecum ingestible device 100 may measure a relatively stable ratio of the measured green reflectance levels to the measured blue reflectance levels. In some embodiments, ingestible device 100 may be configured to determine transitions from the small intestine to the cecum based on these differences. For example, ingestible device 100 may compare recent windows of data to past windows of data, and detect a transition to the cecum in response to determining that the standard deviation of the ratios in the recent window of data is substantially less than the standard deviation of the ratios in the past window of data.

[1075] FIG. 8 is another plot illustrating data collected during an example operation of an ingestible device, which may be used when determining a location of an ingestible device as it transits through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure. Similar to FIG. 7, FIG. 8 may be described in connection with the ingestible device 100 for illustrative purposes. However, this is not intended to be limiting, and plot 800 and data set 802 may be typical of data gathered by any device discussed in this application.

[1076] At 804, shortly after ingestible device 100 begins operation, ingestible device 100 determines that it has reached at least the stomach (e.g., as a result of making a determination similar to the determination discussed in relation to 506 in process 500 (FIG. 5)). Ingestible device 100 continues to gather additional measurements of green and blue reflectance levels (e.g., via sensing sub-unit 126 (FIG. 2)), and at 806 ingestible device 100 determines that a pyloric transition has occurred from the stomach to the duodenum (e.g., as a result of making a determination similar to the determinations discussed in relation to 616-624 of process 600 (FIG. 6)). Notably, the values in data set 802 around 806 jump up precipitously, which is indicative of the higher ratios of measured green reflectance levels to measured blue reflectance levels typical of the duodenum, before falling shortly thereafter. As a result of the reduced values in data set 802, ingestible device 100 determines that a reverse pyloric transition has occurred from the duodenum back to the stomach at 808 (e.g., as a result of making a determination similar to the determinations discussed in relation to 610-612 of process 600 (FIG. 6)). At 810, as a result of the values in data set 802 increasing again, ingestible device 100 determines that another pyloric transition has occurred from the stomach to the duodenum, and shortly thereafter ingestible device 100 proceeds onwards to the jejunum, ileum, and cecum.

[1077] The remainder of the data set 802 depicts the ratios of the measured green reflectance levels to the measured blue reflectance levels throughout the remainder of the GI tract. Notably, at 812, ingestible device reaches the transition point between the ileum and the cecum. As discussed above in relation to FIG. 7, the transition to the cecum is marked by a reduced standard deviation in the ratios of measured green reflectances and measured blue reflectances over time, and ingestible device 100 may be configured to detect a transition to the cecum based on determining that the standard deviation of a recent set of measurements is substantially smaller than the standard deviation of past measurements taken from the jejunum or ileum.

[1078] FIG. 9 is a flowchart of illustrative steps for detecting a transition from a duodenum to a jejunum, which may be used when determining a location of an ingestible device as it transits through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure. Although FIG. 9 may be described in connection with the ingestible device 100 for illustrative purposes, this is not intended to be limiting, and either portions or the entirety of process 900 described in FIG. 9 may be applied to any device discussed in this application (e.g., the ingestible devices 100, 300, and 400), and any of these ingestible devices may be used to perform one or more parts of the process described in FIG. 9. Furthermore, the features of FIG. 9 may be combined with any other systems, methods or processes described in this application. For example, portions of the process described by the process in FIG. 9 may be integrated into the localization process described by FIG. 5 (e.g., as part of 520-524 of process 500 (FIG. 5)). In some embodiments, an ingestible device 100 may perform process 900 while in the duodenum, or in response to detecting entry to the duodenum. In other embodiments, an ingestible device 100 may perform process 900 while in the stomach, or in response to detecting entry into the GI tract. It is also understood that process 900 may be performed in parallel with any other process described in this disclosure (e.g., process 600 (FIG. 6)), which may enable ingestible device 100 to detect entry into various portions of the GI tract, without necessarily detecting entry into a preceding portion of the GI tract. For illustrative purposes, FIG. 9 may be discussed in terms of ingestible device 100 generating and making determinations based on a single set of reflectance levels generated at a single wavelength by a single sensing sub-unit (e.g., sensing sub-unit 126 (FIG. 2)).

[1079] However, it is understood that ingestible device 100 may generate multiple wavelengths of illumination from multiple different sensing sub-units positioned around the circumference of ingestible device (e.g., multiple sensing sub-units positioned at different locations behind window 114 of ingestible device 100 (FIG. 1), and each of the resulting reflectances may be stored as a separate data set. Moreover, each of these sets of reflectance levels may be used to detect muscle contractions by running multiple versions of process 900, each one of which processes data for a different set of reflectances corresponding to data sets obtained from measurements of different wavelengths or measurements made by different sensing sub-units.

[1080] At 902, the ingestible device (e.g., ingestible device 100, 300, or 400) retrieves a set of reflectance levels. For example, ingestible device 100 may retrieve a data set of previously recorded reflectance levels from memory (e.g., from memory circuitry of PCB 120 (FIG. 2)). Each of the reflectance levels may correspond to reflectances previously detected by ingestible device 100 (e.g., via detector 122 (FIG. 2)) from illumination generated by ingestible device 100 (e.g., via illuminator 124 (FIG. 2)), and may represent a value indicative of an amount of light detected in a given reflectance. However, it is understood that any suitable frequency of light may be used, such as light in the infrared, visible, or ultraviolet spectrums. In some embodiments, the reflectance levels may correspond to reflectances previously detected by ingestible device 100 at periodic intervals.

[1081] At 904, the ingestible device (e.g., ingestible device 100, 300, or 400) includes new measurements of reflectance levels in the data set. For example, ingestible device 100 may be configured to detect a new reflectance (e.g., transmit illumination and detect the resulting reflectance using sensing sub-unit 126 (FIG. 2)) at regular intervals, or with sufficient speed as to detect peristaltic waves. For example, ingestible device 100 may be configured to generate illumination and measure the resulting reflectance once every three seconds (i.e., the minimum rate necessary to detect a 0.17 Hz signal), and preferably at a higher rate, as fast at 0.1 second or even faster. It is understood that the periodic interval between measurements may be adapted as needed based on the species of the subject, and the expected frequency of the peristaltic waves to be measured. Every time ingestible device 100 makes a new reflectance level measurement at 904, the new data is included to the data set (e.g., a data set stored within memory circuitry of PCB 120 (FIG. 2)).

[1082] At 906, the ingestible device (e.g., ingestible device 100, 300, or 400) obtains a first subset of recent data by applying a sliding window filter to the data set. For example, ingestible device 100 may retrieve a one-minute worth of data from the data set. If the data set includes values for reflectances measured every second, this would be approximately 60 data points worth of data. Any suitable type of window size may be used, provided that the size of the window is sufficiently large to detect peristaltic waves (e.g., fluctuations on the order of 0.1 Hz to 0.2 Hz for healthy human subjects). In some embodiments, ingestible device 100 may also clean the data, for example, by removing outliers from the first subset of data obtained through the use of the sliding window filter.

[1083] At 908, the ingestible device (e.g., ingestible device 100, 300, or 400) obtains a second subset of recent data by interpolating the first subset of recent data. For example, ingestible device 100 may interpolate the first subset of data in order to generate a second subset of data with a sufficient number of data points (e.g., data points spaced every 0.5 seconds or greater). In some embodiments, this may enable ingestible device 100 to also replace any outlier data points that may have been removed as part of applying the window filter at 906.

[1084] At 910, the ingestible device (e.g., ingestible device 100, 300, or 400) calculates a normalized frequency spectrum from the second subset of data. For example, ingestible device 100 may be configured to perform a fast Fourier transform to convert the second subset of data from a time domain representation into a frequency domain representation. It is understood that depending on the application being used, and the nature of the subset of data, any number of suitable procedures (e.g., Fourier transform procedures) may be used to determine a frequency spectrum for the second subset of data. For example, the sampling frequency and size of the second subset of data may be known in advance, and ingestible device 100 may be configured to have pre-stored values of a normalized discreet Fourier transform (DFT) matrix, or the rows of the DFT matrix corresponding to the 0.1 Hz to 0.2 Hz frequency components of interest, within memory (e.g., memory circuitry of PCB 120 (FIG. 2)). In this case, the ingestible device may use matrix multiplication between the DFT matrix and the data set to generate an appropriate frequency spectrum. An example data set and corresponding frequency spectrum that may be obtained by the ingestible device is discussed in greater detail in relation to FIG. 10.

[1085] At 912, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether at least a portion of the normalized frequency spectrum is between 0.1 Hz and 0.2 Hz above a threshold value of 0.5 Hz. Peristaltic waves in a healthy human subject occur at a rate between 0.1 Hz and 0.2 Hz, and an ingestible device experiencing peristaltic waves (e.g., ingestible device 400 detecting contractions in walls 406 of the jejunum (FIG. 4)) may detect sinusoidal variations in the amplitude of detected reflectances levels that follow a similar 0.1 Hz to 0.2 Hz frequency. If the ingestible device determines that a portion of the normalized frequency spectrum between 0.1 Hz and 0.2 Hz is above a threshold value of 0.5, this measurement may be consistent with peristaltic waves in a healthy human subject, and process 900 proceeds to 914 where ingestible device 100 stores data indicating a muscle contraction was detected. Alternatively, if the ingestible device determines that no portion of the normalized frequency spectrum between 0.1 Hz and 0.2 Hz above a threshold value of 0.5, process 900 proceeds directly to 904 to make new measurements and to continue to monitor for new muscle contractions. It is understood that a threshold value other than 0.5 may be used, and that the exact threshold may depend on the sampling frequency and type of frequency spectrum used by ingestible device 100.

[1086] At 914, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating a muscle contraction was detected. For example, ingestible device 100 may store data in memory (e.g., memory circuitry of PCB 120 (FIG. 2)) indicating that a muscle contraction was detected, and indicating the time that the muscle contraction was detected. In some embodiments, ingestible device 100 may also monitor the total number of muscle contractions detected, or the number of muscle contractions detected in a given time frame. In some embodiments, detecting a particular number of muscle contractions may be consistent with ingestible device 100 being within the jejunum (e.g., jejunum 314 (FIG. 3)) of a healthy human subject. After detecting a muscle contraction, process 900 proceeds to 916.

[1087] At 916, the ingestible device (e.g., ingestible device 100, 300, or 400) determines whether a total number of muscle contractions exceeds a predetermined threshold number. For example, ingestible device 100 may retrieve the total number of muscle contractions detected from memory (e.g., from memory circuitry of PCB 120 (FIG. 2)), and compare the total number to a threshold value. In some embodiments, the threshold value may be one, or any number larger than one. The larger the threshold value, the more muscle contractions need to be detected before ingestible device 100 stores data indicating that it has entered the jejunum. In practice, setting the threshold value as three or higher may prevent the ingestible device from detecting false positives (e.g., due to natural movement of the GI tract organs, or due to movement of the subject). If the total number of contractions exceeds the predetermined threshold number, process 900 proceeds to 918 to store data indicating detection of a transition from the duodenum to the jejunum. Alternatively, if the total number of contractions does not exceed a predetermined threshold number, process 900 proceeds to 904 to include new measurements of reflectance levels in the data set. An example plot of the muscle contractions detected over time is discussed in greater detail in relation to FIG. 11.

[1088] At 918, the ingestible device (e.g., ingestible device 100, 300, or 400) stores data indicating detection of a transition from the duodenum to the jejunum. For example, ingestible device 100 may store data in memory (e.g., from memory circuitry of PCB 120 (FIG. 2)) indicating that the jejunum has been reached. In some embodiments, if ingestible device 100 is configured to perform all or part of process 900 while in the stomach, ingestible device 100 may store data at 918 indicating detection of a transition from the stomach directly to the jejunum (e.g., as a result of transitioning too quickly through the duodenum for the pyloric transition to be detected using process 600 (FIG. 6)).

[1089] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may be configured to obtain a fluid sample from the environment external to a housing of the ingestible device in response to identifying a change in the location of the ingestible device. For example, ingestible device 100 may be configured to obtain a fluid sample from the environment external to the housing of ingestible device 100 (e.g., through the use of optional opening 116 and optional rotating assembly 118 (FIG. 2)) in response to determining that the ingestible device is located within the jejunum (e.g., jejunum 314 (FIG. 3)). In some embodiments, ingestible device 100 may also be equipped with appropriate diagnostics to detect certain medical conditions based on the retrieved fluid sample, such as small intestinal bacterial overgrowth (SIBO).

[1090] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may be configured to deliver a dispensable substance that is pre-stored within the ingestible device from the ingestible device into the gastrointestinal tract in response to identifying the change in the location of the ingestible device. For example, ingestible device 100 may have a dispensable substance pre-stored within the ingestible device 100 (e.g., within a storage chamber or cavity on optional storage sub-unit 118-3 (FIG. 2)), and ingestible device 100 may be configured to dispense the substance into the gastrointestinal tract (e.g., through the use of optional opening 116 and optional rotating assembly 118 (FIG. 2)) when the ingestible device 100 detects that the ingestible device 100 is located within the jejunum (e.g., jejunum 314 (FIG. 3)). In some embodiments, this may enable ingestible device 100 to deliver substances (e.g., therapeutics and medicaments) at targeted locations within the GI tract.

[1091] In some embodiments, the ingestible device (e.g., ingestible device 100, 300, or 400) may be configured to perform an action based on the total number of detected muscle contractions. For example, ingestible device 100 may be configured to retrieve data indicative of the total number of muscle contractions (e.g., from memory circuitry of PCB 120 (FIG. 2)), and compare that to an expected number muscle contractions in a healthy individual. In response, the ingestible device may either dispense a substance into the gastrointestinal tract (e.g., through the use of optional opening 116 and optional rotating assembly 118 (FIG. 2)), or may obtain a fluid sample from the environment external to the housing of ingestible device 100 (e.g., through the use of optional opening 116 and optional rotating assembly 118 (FIG. 2)). For instance, ingestible device 100 may be configured to obtain a sample in response to determining that a number of detected muscle contractions is abnormal, and differs greatly from the expected number. As another example, ingestible device 100 may be configured to deliver a substance into the GI tract (such as a medicament), in response to determining that the detected muscle contractions are consistent with a functioning GI tract in a healthy individual.

[1092] It will be understood that the steps and descriptions of the flowcharts of this disclosure, including FIG. 9, are merely illustrative. Any of the steps and descriptions of the flowcharts, including FIG. 9, may be modified, omitted, rearranged, performed in alternate orders or in parallel, two or more of the steps may be combined, or any additional steps may be added, without departing from the scope of the present disclosure. For example, the ingestible device 100 may calculate the mean and the standard deviation of multiple data sets in parallel (e.g., multiple data sets, each one corresponding to a different wavelength of reflectance or different sensing sub-unit used to detect the reflectance) in order to speed up the overall computation time. Furthermore, it should be noted that the steps and descriptions of FIG. 9 may be combined with any other system, device, or method described in this application, and any of the ingestible devices or systems discussed in this application could be used to perform one or more of the steps in FIG. 9.

[1093] FIG. 10 is a plot illustrating data collected during an example operation of an ingestible device, which may be used when detecting a transition from a duodenum to a jejunum, in accordance with some embodiments of the disclosure. Diagram 1000 depicts a time domain plot 1002 of a data set of reflectance levels measured by an ingestible device (e.g., the second subset of data discussed in relation to 908 of FIG. 9). In some embodiments, ingestible device 100 may be configured to gather data points at semi-regular intervals approximately 0.5 seconds apart. By comparison, diagram 1050 depicts a frequency domain plot 1004 of the same data set of reflectance levels measured by an ingestible device (e.g., as a result of ingestible device 100 calculating a frequency spectrum at 910 of FIG. 9). In some embodiments, ingestible device 100 may be configured to calculate the frequency spectrum through any convenient means.

[1094] In diagram 1050, the range of frequencies 1006 between 0.1 Hz and 0.2 Hz may be the range of frequencies that ingestible device 100 searches in order to detect muscle contractions. As shown in diagram 1050, there is a strong peak in the frequency domain plot 1004 around 0.14 Hz, which is consistent with the frequency of peristaltic motion in a healthy human individual. In this case, an ingestible device 100 analyzing frequency domain plot 1004 may be configured to determine that the data is consistent with a detected muscle contraction (e.g., using a process similar to 912 of process 900 (FIG. 9)), and may store data (e.g., in memory circuitry of PCB 120 (FIG. 2)) indicating that a muscle contraction has been detected. Because the muscle contraction was detected from the one-minute window of data ending at 118 minutes, ingestible device 100 may also store data indicating that the muscle contraction was detected at the 118-minute mark (i.e., which may indicate that the ingestible device 100 was turned on and ingested by the subject 118 minutes ago).

[1095] FIG. 11 is a plot illustrating muscle contractions detected by an ingestible device over time, which may be used when determining a location of an ingestible device as it transits through a gastrointestinal (GI) tract, in accordance with some embodiments of the disclosure. In some embodiments, ingestible device 100 may be configured to detect muscle contractions, and store data indicative of when each muscle contraction is detected (e.g., as part of 914 of process 900 (FIG. 9)). Plot 1100 depicts the detected muscle contractions 1106 over time, with each muscle contraction being represented by a vertical line reaching from O to 1 on the y-axis.

[1096] At 1102, around the 10-minute mark, ingestible device 100 first enters the duodenum (e.g., as determined by ingestible device 100 performing process 600 (FIG. 6)). Shortly thereafter, at 1108, ingestible device 100 begins to detect several muscle contractions 1106 in quick succession, which may be indicative of the strong peristaltic waves that form in the jejunum (e.g., jejunum 314 (FIG. 3)). Later, around 1110, ingestible device 100 continues to detect intermittent muscle contractions, which may be consistent with an ingestible device 100 within the ileum. Finally at 1104, ingestible device 100 transitions out of the small intestine, and into the cecum. Notably, ingestible device 100 detects more frequent muscle contractions in the jejunum portion of the small intestine as compared to the ileum portion of the small intestine, and ingestible device 100 does not measure any muscle contractions after having exited the small intestine. In some embodiments, ingestible device 100 may incorporate this information into a localization process. For example, ingestible device 100 may be configured to detect a transition from a jejunum to an ileum in response to determining that a frequency of detected muscle contractions (e.g., the number of muscle contractions measured in a given 10-minute window) has fallen below a threshold number. As another example, ingestible device 100 may be configured to detect a transition from an ileum to a cecum in response to determining that no muscle contractions have been detected for a threshold period of time. It is understood that these examples are intended to be illustrative, and not limiting, and that measurements of muscle contractions may be combined with any of the other processes, systems, or methods discussed in this disclosure.

[1097] FIG. 12 is a flowchart 1200 for certain embodiments for determining a transition of the device from the jejunum to the ileum. It is to be noted that, in general, the jejunum is redder and more vascular than the ileum. Moreover, generally, in comparison to the ileum, the jejunum has a thicker intestine wall with more messentary fat. These differences between the jejunum and the ileum are expected to result in differences in optical responses in the jejunum relative to the ileum. Optionally, one or more optical signals may be used to investigate the differences in optical responses. For example, the process can include monitoring a change in optical response in reflected red light, blue light, green light, ratio of red light to green light, ratio of red light to blue light, and/or ratio of green light to blue light. In some embodiments, reflected red light is detected in the process.

[1098] Flowchart 1200 represents a single sliding window process. In step 1210, the jejunum reference signal is determined based on optical reflection. Typically, this signal is as the average signal (e.g., reflected red light) over a period of time since the device was determined to enter the jejunum. The period of time can be, for example, from five minutes to 40 minutes (e.g., from 10 minutes to 30 minutes, from 15 minutes to 25 minutes). In step 1220, the detected signal (e.g., reflected red light) just after the period of time used in step 1210 is normalized to the reference signal determined in step 1210. In step 1230, the signal (e.g., reflected red light) is detected. In step 1240, the mean signal detected based on the single sliding window is compared to a signal threshold. The signal threshold in step 1240 is generally a fraction of the reference signal of the jejunum reference signal determined in step 1210. For example, the signal threshold can be from 60% to 90% (e.g., from 70% to 80%) of the jejunum reference signal. If the mean signal exceeds the signal threshold, then the process determines that the device has entered the ileum at step 1250. If the mean signal does not exceed the signal threshold, then the process returns to step 1230.

[1099] FIG. 13 is a flowchart 1200 for certain embodiments for determining a transition of the device from the jejunum to the ileum using a two sliding window process. In step 1310, the jejunum reference signal is determined based on optical reflection. Typically, this signal is as the average signal (e.g., reflected red light) over a period of time since the device was determined to enter the jejunum. The period of time can be, for example, from five minutes to 40 minutes (e.g., from 10 minutes to 30 minutes, from 15 minutes to 25 minutes). In step 1320, the detected signal (e.g., reflected red light) just after the period of time used in step 1310 is normalized to the reference signal determined in step 1310. In step 1330, the signal (e.g., reflected red light) is detected. In step 1340, the mean difference in the signal detected based on the two sliding windows is compared to a signal threshold. The signal threshold in step 1340 is based on whether the mean difference in the detected signal exceeds a multiple (e.g., from 1.5 times to five times, from two times to four times) of the detected signal of the first window. If signal threshold is exceeded, then the process determines that the device has entered the ileum at step 1350. If the signal threshold is not exceeded, then the process returns to step 1330.

[1100] FIG. 14 is a flowchart 1400 for a process for certain embodiments for determining a transition of the device from the ileum to the cecum. In general, the process involves detecting changes in the reflected optical signal (e.g., red light, blue light, green light, ratio of red light to green light, ratio of red light to blue light, and/or ratio of green light to blue light). In some embodiments, the process includes detecting changes in the ratio of reflected red light to reflected green light, and also detecting changes in the ratio of reflected green light to reflected blue light. Generally, in the process 1400, the sliding window analysis (first and second windows) discussed with respect to process 600 is continued.

[1101] Step 1410 includes setting a first threshold in a detected signal, e.g., ratio of detected red light to detected green light, and setting a second threshold for the coefficient of variation for a detected signal, e.g., the coefficient of variation for the ratio of detected green light to detected blue light. The first threshold can beset to a fraction (e.g., from 0.5 to 0.9, from 0.6 to 0.8) of the average signal (e.g., ratio of detected red light to detected green light) in the first window, or a fraction (e.g., from 0.4 to 0.8, from 0.5 to 0.7) of the mean difference between the detected signal (e.g., ratio of detected red light to detected green light) in the two windows. The second threshold can be set to 0.1 (e.g., 0.05, 0.02).

[1102] Step 1420 includes detecting the signals in the first and second windows that are to be used for comparing to the first and second thresholds.

[1103] Step 1430 includes comparing the detected signals to the first and second thresholds. If the corresponding value is not below the first threshold or the corresponding value is not below the second threshold, then it is determined that the device has not left the ileum and entered the cecum, and the process returns to step 1420. If the corresponding value is below the first threshold and the corresponding value is below the second threshold, then it is determined that the device has left the ileum and entered the cecum, and the proceeds to step 1440.

[1104] Step 1450 includes determining whether it is the first time that that the device was determined to leave the ileum and enter the cecum. If it is the first time that the device was determined to leave the ileum and enter the cecum, then the process proceeds to step 1460. If it is not the first time that the device has left the ileum and entered the cecum, then the process proceeds to step 1470.

[1105] Step 1460 includes setting a reference signal. In this step the optical signal (e.g., ratio of detected red light to detected green light) as a reference signal.

[1106] Step 1470 includes determining whether the device may have left the cecum and returned to the ileum. The device is determined to have left the cecum and returned to the ileum if the corresponding detected signal (e.g., ratio of detected red light to detected green light) is statistically comparable to the reference signal (determined in step 1460) and the coefficient of variation for the corresponding detected signal (e.g., ratio of detected green light to detected blue light) exceeds the second threshold. If it is determined that the device may have left the cecum and returned to the ileum, the process proceeds to step 1480.

[1107] Step 1480 includes continuing to detect the relevant optical signals for a period of time (e.g., at least one minute, from five minutes to 15 minutes).

[1108] Step 1490 includes determining whether the signals determined in step 1480 indicate (using the methodology discussed in step 1470) that the device re-entered the ileum. If the signals indicate that the device re-entered the ileum, the process proceeds to step 1420. If the signals indicate that the device is in the cecum, the process proceeds to step 1492.

[1109] Step 1492 includes continuing to monitor the relevant optical signals for a period of time (e.g., at least 30 minutes, at least one hour, at least two hours).

[1110] Step 1494 includes determining whether the signals determined in step 1492 indicate (using the methodology discussed in step 1470) that the device re-entered the ileum. If the signals indicate that the device re-entered the ileum, the process proceeds to step 1420. If the signals indicate that the device is in the cecum, the process proceeds to step 1496.

[1111] At step 1496, the process determines that the device is in the cecum.

[1112] FIG. 15 is a flowchart 1500 for a process for certain embodiments for determining a transition of the device from the cecum to the colon. In general, the process involves detecting changes in the reflected optical signal (e.g., red light, blue light, green light, ratio of red light to green light, ratio of red light to blue light, and/or ratio of green light to blue light). In some embodiments, the process includes detecting changes in the ratio of reflected red light to reflected green light, and also detecting changes in the ratio of reflected blue light. Generally, in the process 1500, the sliding window analysis (first and second windows) discussed with respect to process 1400 is continued.

[1113] In step 1510, optical signals (e.g., the ratio of reflected red signal to reflected green signal, and reflected blue signal) are collected for a period of time (e.g., at least one minute, at least five minutes, at least 10 minutes) while the device is in the cecum (e.g., during step 1480). The average values for the recorded optical signals (e.g., the ratio of reflected red signal to reflected green signal, and reflected blue signal) establish the cecum reference signals.

[1114] In step 1520, the optical signals are detected after it has been determined that the device entered the cecum (e.g., at step 1440). The optical signals are normalized to the cecum reference signals.

[1115] Step 1530 involves determining whether the device has entered the colon. This includes determining whether any of three different criteria are satisfied. The first criterion is satisfied if the mean difference in the ratio of a detected optical signal (e.g., ratio of detected red signal to the detected green) is a multiple greater than one (e.g., 2X, 3X, 4X) the standard deviation of the corresponding signal (e.g., ratio of detected red signal to the detected green) in the second window. The second criterion is satisfied if the mean of a detected optical signal (e.g., a ratio of detected red light to detected green light) exceeds a given value (e.g., exceeds one). The third criterion is satisfied if the coefficient of variation of an optical signal (e.g., detected blue light) in the first window exceeds a given value (e.g., exceeds 0.2). If any of the three criteria are satisfied, then the process proceeds to step 1540. Otherwise, none of the three criteria are satisfied, the process returns to step 1520.

[1116] For illustrative purposes the disclosure focuses primarily on a number of different example embodiments of an ingestible device, and example embodiments of methods for determining a location of an ingestible device within a GI tract. However, the possible ingestible devices that may be constructed are not limited to these embodiments, and variations in the shape and design may be made without significantly changing the functions and operations of the device. Similarly, the possible procedures for determining a location of the ingestible device within the GI tract are not limited to the specific procedures and embodiments discussed (e.g., process 500 (FIG. 5), process 600 (FIG. 6), process 900 (FIG. 9), process 1200 (FIG. 12), process 1300 (FIG. 13), process 1400 (FIG. 14) and process 1500 (FIG. 15)). Also, the applications of the ingestible devices described herein are not limited merely to gathering data, sampling and testing portions of the gastrointestinal tract, or delivering medicament. For example, in some embodiments the ingestible device may be adapted to include a number of chemical, electrical, or optical diagnostics for diagnosing a number of diseases. Similarly, a number of different sensors for measuring bodily phenomenon or other physiological qualities may be included on the ingestible device. For example, the ingestible device may be adapted to measure elevated levels of certain chemical compounds or impurities in the gastrointestinal tract, or the combination of localization, sampling, and appropriate diagnostic and assay techniques incorporated into a sampling chamber may be particularly well suited to determine the presence of small intestinal bacterial overgrowth (SIBO).

[1117] At least some of the elements of the various embodiments of the ingestible device described herein that are implemented via software (e.g., software executed by control circuitry within PCB 120 (FIG. 2)) may be written in a high-level procedural language such as object oriented programming, a scripting language or both. Accordingly, the program code may be written in C, C* or any other suitable programming language and may comprise modules or classes, as is known to those skilled in object oriented programming. Alternatively, or in addition, at least some of the elements of the embodiments of the ingestible device described herein that are implemented via software may be written in assembly language, machine language or firmware as needed. In either case, the language may be a compiled or an interpreted language.

[1118] At least some of the program code used to implement the ingestible device can be stored on a storage media or on a computer readable medium that is readable by a general or special purpose programmable computing device having a processor, an operating system and the associated hardware and software that is necessary to implement the functionality of at least one of the embodiments described herein. The program code, when read by the computing device, configures the computing device to operate in a new, specific and predefined manner in order to perform at least one of the methods described herein.

[1119] Furthermore, at least some of the programs associated with the systems, devices, and methods of the example embodiments described herein are capable of being distributed in a computer program product comprising a computer readable medium that bears computer usable instructions for one or more processors. The medium may be provided in various forms, including non-transitory forms such as, but not limited to, one or more diskettes, compact disks, tapes, chips, and magnetic and electronic storage. In some embodiments, the medium may be transitory in nature such as, but not limited to, wire-line transmissions, satellite transmissions, internet transmissions (e.g. downloads), media, digital and analog signals, and the like. The computer useable instructions may also be in various formats, including compiled and non-compiled code.

[1120] The techniques described above can be implemented using software for execution on a computer. For instance, the software forms procedures in one or more computer programs that execute on one or more programmed or programmable computer systems (which may be of various architectures such as distributed, client/server, or grid) each including at least one processor, at least one data storage system (including volatile and non-volatile memory and/or storage elements), at least one input device or port, and at least one output device or port.

[1121] The software may be provided on a storage medium, such as a CD-ROM, readable by a general or special purpose programmable computer or delivered (encoded in a propagated signal) over a communication medium of a network to the computer where it is executed. All of the functions may be performed on a special purpose computer, or using special-purpose hardware, such as coprocessors. The software may be implemented in a distributed manner in which different parts of the computation specified by the software are performed by different computers. Each such computer program is preferably stored on or downloaded to a storage media or device (e.g., solid state memory or media, or magnetic or optical media) readable by a general or special purpose programmable computer, for configuring and operating the computer when the storage media or device is read by the computer system to perform the procedures described herein. The inventive system may also be considered to be implemented as a computer-readable storage medium, configured with a computer program, where the storage medium so configured causes a computer system to operate in a specific and predefined manner to perform the functions described herein.

Methods and Mechanisms of Delivery

[1122] FIG. 16 provides an example mock-up diagram illustrating aspects of a structure of an ingestible device 1600 for delivering a dispensable substance, such as a formulation of a therapeutic agent described herein, according to some embodiments described herein. In some embodiments, the ingestible device 1600 may generally be in the shape of a capsule, a pill or any swallowable form that may be orally consumed by an individual. In this way, the ingestible device 1600 may be ingested by a patient and may be prescribed by healthcare practitioners and patients.

[1123] FIG. 16 provides an example mock-up diagram illustrating aspects of a structure of an ingestible device 1600 for delivering a dispensable substance, according to some embodiments described herein. In some embodiments, the ingestible device 1600 may generally be in the shape of a capsule, a pill or any swallowable form that may be orally consumed by an individual. In this way, the ingestible device 1600 may be ingested by a patient and may be prescribed by healthcare practitioners and patients.

[1124] The ingestible device 1600 includes a housing 1601 that may take a shape similar to a capsule, a pill, and/or the like, which may include two ends 1602a-b. The housing 1601 may be designed to withstand the chemical and mechanical environment of the GI tract (e.g., effects of muscle contractile forces and concentrated hydrochloric acid in the stomach). A broad range of materials that may be used for the housing 1601. Examples of these materials include, but are not limited to, thermoplastics, fluoropolymers, elastomers, stainless steel and glass complying with ISO 10993 and USP Class VI specifications for biocompatibility; and any other suitable materials and combinations thereof.

[1125] In some embodiment, the wall of the housing 1601 may have a thickness of 0.5 mm-1 mm, which is sufficient to sustain an internal explosion (e.g., caused by hydrogen ignition or over pressure inside the housing).

[1126] The housing 1601 may or may not have a pH-sensitive enteric coating to detect or otherwise be sensitive to a pH level of the environment external to the ingestible device. As discussed elsewhere in the application in more detail, the ingestible device 1600 may additionally or alternatively include one more sensors, e.g., temperature sensor, optical sense.

[1127] The housing 1601 may be formed by coupling two enclosure portions together. The ingestible device 1600 may include an electronic component within the housing 1600. The electronic component may be placed proximally to an end 1602b of the housing, and includes a printed circuit board (PCB), a battery, an optical sensing unit, and/or the like.

[1128] The ingestible device 1600 further includes a gas generating cell 1603 that is configured to generate gas and thus cause an internal pressure within the housing 1601. In some embodiments, the gas generating cell may include or be connected to a separate channel or valve of the ingestible device such that gas may be release through the channel or valve to create a motion to alter the position of the ingestible device within the GI tract. Such gas release can also be used to position the ingestible device relative to the intestinal lining. In another embodiment, gas may be released through the separate channel or valve to alter the surface orientation of the intestinal tissue prior to delivery of the dispensable substance.

[1129] A traveling plunger 1604 may be placed on top of the gas generating cell 1603 within the housing 1601. The traveling plunger 1604 is a membrane that separates the gas generating cell 1603 and a storage reservoir that stores the dispensable substance 1605. In some embodiments, the traveling plunger 1604 may be a movable piston. In some embodiments, the traveling plunger 1604 may instead be a flexible membrane such as but not limited to a diaphragm. In some embodiments, the traveling plunger 1604, which may have the form of a flexible diaphragm, may be placed along an axial direction of the housing 1601, instead of being placed on top of the gas generating cell 1603. The traveling plunger or the membrane 1604 may move (when the membrane 1604 is a piston) or deform (when the membrane 1604 is a diaphragm) towards a direction of the end 1602a of the housing, when the gas generating cell 1603 generates gas to create an internal pressure that pushes the membrane 1604. In this way, the membrane or traveling plunger 1604 may push the dispensable substance 1605 out of the housing via a dispensing outlet 1607.

[1130] The housing 1601 may include a storage reservoir storing one or more dispensable substances 1605 adjacent to the traveling plunger 1604. The dispensable substance 1605 may be a therapeutic or medical agent that may take a form of a powder, a compressed powder, a fluid, a semi-liquid gel, or any other dispensable or deliverable form. The delivery of the dispensable substance 1605 may take a form such as but not limited to bolus, semi-bolus, continuous, burst drug delivery, and/or the like. In some embodiments, a single bolus is delivered proximate to the disease location. In some embodiments, more than one bolus is released at one location or more than one location. In some embodiments the release of more than one bolus is triggered according to a pre-programmed algorithm. In some embodiments the release profile is continuous. In some embodiments the release profile is time-based. In some embodiments the release profile is location-based. In some embodiments, the amount delivered is based on the severity and/or extent of the disease in the following manner. In some embodiments, the bolus is delivered in one or more of the following locations: stomach; duodenum; proximal jejunum; ileum; cecum; ascending colon; transverse colon; descending colon.

[1131] In some embodiments the dispensable substance is a small molecule therapeutic that is released in the cecum and/or other parts of the large intestine. Small molecules that are administered by typical oral routes are primarily absorbed in the small intestine, with much lower absorption taking place in the large intestine (outside of the rectum). Accordingly, an ingestible device that is capable of releasing a small molecule selectively in the large intestine (e.g., the cecum) with resulting low systemic levels (even when high doses are used) is attractive for subjects with inflammatory bowel disease in the large intestine.

[1132] In some embodiments, the storage reservoir may include multiple chambers, and each chamber stores a different dispensable substance. For example, the different dispensable substances can be released at the same time via the dispensing outlet 1607. Alternatively, the multiple chambers may take a form of different layers within the storage reservoir such that the different dispensable substance from each chamber is delivered sequentially in an order. In one example, each of the multiple chambers is controlled by a separate traveling plunger, which may be propelled by gas generation. The electronic component may control the gas generating cell 1603 to generate gas to propel a specific traveling plunger, e.g., via a separate gas generation chamber, etc., to delivery the respective substance. In some embodiments, the content of the multiple chambers may be mixed or combined prior to release, for example, to activate the drug.

[1133] The ingestible device 1600 may include a dispensing outlet 1607 at one end 1602a of the housing 1601 to direct the dispensable substance 105 out of the housing. The dispensing outlet 1607 may include an exit valve, a slit or a hole, a jet injection nozzle with a syringe, and/or the like. When the traveling plunger 1604 moves towards the end 1602a of the housing 1601, an internal pressure within the storage reservoir may increase and push the dispensing outlet to be open to let the dispensable substance 1605 be released out of the housing 1601.

[1134] In an embodiment, a pressure relief device 1606 may be placed within the housing 1601, e.g., at the end 1602a of the housing 1601.

[1135] In some embodiments, the housing 1601 may include small holes (e.g., with a diameter smaller than 2 mm), e.g., on the side of the housing 1601, or at the end 1602a to facilitate loading the dispensable substance into the storage reservoir.

[1136] In some embodiments, a feedback control circuit (e.g., a feedback resistor, etc.) may be added to send feedback from the gas generating cell 1603 to the electronic component such that when the internal pressure reaches a threshold level, the electronic component may control the gas generating cell 1603 to turn off gas generation, or to activate other safety mechanism (e.g., feedback-controlled release valve, etc.). For example, an internal pressure sensor may be used to measure the internal pressure within the ingestible device and generate feedback to the feedback control circuit.

[1137] FIG. 17 provides an example diagram illustrating aspects of a mechanism for a gas generating cell 1603 configured to generate a gas to dispense a substance, according to some embodiments described herein. As shown in FIG. 17, the gas generating cell 1603 generates a gas 1611 which can propel the dispensable substance 1605 out of the dispensing outlet 1607. A variable resistor 1608 may be connected to a circuit with the gas generating cell 1603 such that the variable resistor 1608 may be used to control an intensity and/or an amount of gas 1611 (e.g., hydrogen) generated by the cell 1603. Specifically, the gas generating cell 1603 may be a battery form factor cell that is capable of generating hydrogen when a resistor is applied. In this way, as the gas generating cell 1603 only needs the use of a resistor only without any active power requirements, the gas generating cell 1603 may be integrated into an ingestible device such as a capsule with limited energy/power available. For example, the gas generating cell 1603 may be compatible with a capsule at a size of 26 mm13 mm or smaller.

[1138] In some embodiments, based on the elution rate of gas from the cell, and an internal volume of the ingestible device, it may take time to generate sufficient gas 1611 to deliver the substance 1605, and the time required may be 30 seconds or longer. For example, the time to generate a volume of hydrogen equivalent to 500 L of fluid would be approximately 5 minutes. A longer period of time may be needed based upon non-ideal conditions within the ingestible device, such as friction, etc. Thus, given that the production of gas (e.g., hydrogen) may take time, gas generation may need to start prior to the ingestible device arriving at the site of delivery to build pressure up within the device. The ingestible device may then need to know when it is approaching the site of delivery. For example, the device may start producing gas on an entry transition, which is determined by temperature, so as to produce enough gas to be close to the pressure high enough to deliver the dispensable substance. The ingestible device may then only start producing gas again when it arrives at the site of delivery, which will cause the internal pressure within the ingestible device to reach a level required by the dispensing outlet to release the dispensable substance. Also, for regio-specific delivery, the ingestible device may estimate the time it takes to build up enough pressure to deliver the dispensable substance before the ingestible device arrives at a specific location, to activate gas generation.

[1139] For example, for systemic delivery, when an internal volume of the ingestible device is around 500 L, a gas generation time of 2 hours, an initial pressure of approximately 300 pound per square inch absolute (psia) may be generated, with higher and lower pressures possible. The generated pressure may drop when air enters the storage reservoir which was previously occupied by the dispensable substance during the dispensing process. For systemic drug delivery, a force with a generated pressure of approximately 100 to 360 pound per square inch (psi) may be required for dermal penetration, e.g., to penetrate the mucosa or epithelial layer. The pressure may also vary depending on the nozzle design at the dispensing outlet, fluid viscosity, and surrounding tissue proximity and properties.

[1140] The gas 1611 that may be generated for a continuous delivery of drug (e.g., 1 cc H.sub.2 in 4 hours, 16 breaths per minute at 0.5L tidal volume) may equate to 1 cc hydrogen in approximately 2000L of exhaled air, or approximately 0.5 ppm H2, which is below physiologic values of exhaled hydrogen. Reducing this time to 10 minutes equates to approximately 13 ppm hydrogen. Thus, due to the length of intestine that may be covered during this time period, the ingestible device may possess a higher localized value than physiologic.

[1141] FIGS. 18 and 19, disclosed in U.S. Provisional Application No. 62/385,553, incorporated by reference herein in its entirety, illustrates an example of an ingestible device for localized delivery of pharmaceutical compositions disclosed herein, in accordance with particular implementations. The ingestible device 1600 includes a piston or drive element 1634 to push for drug delivery, in accordance with particular implementations described herein. The ingestible device 1600 may have one or more batteries 1631 placed at one end 1602a of a housing 1601 to provide power for the ingestible device 1600. A printed circuit board (PCB) 1632 may be placed adjacent to a battery or other power source 1631, and a gas generating cell 1603 may be mounted on or above the PCB 1632. The gas generating cell 1603 may be sealed from the bottom chamber (e.g., space including 1631 and 1632) of the ingestible device 1600. A movable piston 1634 may be placed adjacent to the gas generating cell 1603. In this way, gas generation from the gas generating cell 1603 may propel a piston 1634 to move towards another end 1602b of the housing 1601 such that the dispensable substance in a reservoir compartment 1635 can be pushed out of the housing through a dispensing outlet 1607, e.g., the movement is shown at 1636, with the piston 1634 at a position after dispensing the substance. The dispensing outlet 1607 may comprise a plug. The reservoir compartment 1635 can store the dispensable substance (e.g., drug substance), or alternatively the reservoir compartment can house a storage reservoir 1661 which comprises the dispensable substance. The reservoir compartment 1635 or storage reservoir 1661 may have a volume of approximately 600 L or even more dispensable substance, which may be dispensed in a single bolus, or gradually over a period of time.

[1142] The battery cells 1631 may have a height of 1.65 mm each, and one to three batteries may be used. The height of the piston may be reduced with custom molded part for around 1.5 mm to save space. If the gas generating cell 1603 is integrated with the piston 1634, the overall height of the PCB, batteries and gas generating cell in total can be reduced to around 5 mm, thus providing more space for drug storage. For example, for an ingestible device of 7.8 mm in length (e.g., from end 1602a to the other end 1602b), a reservoir compartment 1635 or a storage reservoir 1661 of approximately 600 L may be used for drug delivery. For another example, for an ingestible device of 17.5 mm in length, a reservoir compartment 1635 or a storage reservoir 1661 of approximately 1300 L may be used for drug release.

[1143] In some implementations, at the reservoir 1635 or 1661 for storing a therapeutically effective amount of any of the agents described herein at least a portion of the device housing 1601. The therapeutically effective amount of the any of the agents described herein can be stored in the reservoir 1635 or 1661 at a particular pressure, for example, determined to be higher than a pressure inside the GI tract so that once the reservoir 1635 or 1661 is in fluid communication with the GI tract, the =agent is automatically released. In certain implementations, the reservoir compartment 1635 includes a plurality of chambers, and each of the plurality of the chambers stores a different dispensable substance or a different storage reservoir 1661.

[1144] In certain embodiments, the storage reservoir 1661 is a compressible component or has compressible side walls. In particular embodiments, the compressible component can be composed, at least in part, or coated (e.g., internally) with polyvinyl chloride (PVC), silicone, DEHP (di-2-ethylhexyl phthalate), Tyvek, polyester film, polyolefin, polyethylene, polyurethane, or other materials that inhibit the immune modulator (e.g., any of the immune modulators described herein) from sticking to the reservoir and provide a sterile reservoir environment for the immune modulator. The storage reservoir 1661 can be hermetically sealed. The reservoir compartment 1635 or storage reservoir 1661 can be configured to store the immune modulator (e.g., any of the immune modulators described herein) in quantities in the range of 0.01 mL-2 mL, such as 0.05 mL-2 mL, such as 0.05 mL-2 mL, such as 0.6 mL-2 mL. In some embodiments, the storage reservoir 1661 is attachable to the device housing 1601, for example, in the reservoir compartment. Accordingly, the storage reservoir 1635 can be loaded with the immune modulator (e.g., any of the immune modulators described herein) prior to being positioned in and/or coupled to the ingestible device housing 1601. The ingestible device housing 1601 includes one or more openings configured as a loading port to load the dispensable substance into the reservoir compartment. In another embodiment, the ingestible device housing 1601 includes one or more openings configured as a vent.

[1145] In certain embodiments, the ingestible device housing 1601 includes one or more actuation systems (e.g., gas generating cell 1603) for pumping the immune modulator (e.g., any of the immune modulators described herein) from the reservoir 1635. In some embodiments, the actuation system can include a mechanical, electrical, electromechanical, hydraulic, and/or fluid actuation system. For example, a chemical actuation means may use chemical reaction of mixing one or more reagents to generate a sufficient volume of gas to propel the piston or drive element 1634 for drug release. The actuation system can be integrated into the reservoir compartment 1635 or can be an auxiliary system acting on or outside of the reservoir compartment 1635. For example, the actuation system can include pumping system for pushing/pulling the immune modulator (e.g., any of the immune modulators described herein) out of the reservoir compartment 1635 or the actuation system can be configured to cause the reservoir compartment 1635 to change structurally so that the volume inside of the reservoir compartment 1635 changes, thereby dispensing the immune modulator from the reservoir compartment 1635. The actuation system can include an energy storage component such as a battery or a capacitor for powering the actuation system. The actuation system can be actuated via gas pressure or a system storing potential energy, such as energy from an elastic reservoir component being expanded during loading of the reservoir and after being positioned in the ingestible device housing 1601 being subsequently released from the expanded state when the ingestible device housing is at the location for release within the GI tract. In certain embodiments, the reservoir compartment 1635 can include a membrane portion, whereby the immune modulator (e.g., any of the immune modulators described herein) is dispensed from the reservoir compartment 1635 or storage reservoir 1661 via osmotic pressure.

[1146] In particular embodiments the storage reservoir 1661 is in a form of a bellow that is configured to be compressed via a pressure from the gas generating cell. The immune modulator may be loaded into the bellow, which may be compressed by gas generation from the gas generating cell or other actuation means to dispense the dispensable substance through the dispensing outlet 1607 and out of the housing 1601. In some embodiments, the ingestible device includes a capillary plate placed between the gas generating cell and the first end of the housing, and a wax seal between the gas generating cell and the reservoir, wherein the wax seal is configured to melt and the dispensable substance is pushed through the capillary plate by a pressure from the gas generating cell. The shape of the bellow may aid in controlled delivery. The reservoir compartment 1635 includes a dispensing outlet, such as a valve or dome slit 1662 extending out of an end of the housing 1601, in accordance with particular implementations. Thus when the bellow is being compressed, the dispensable substance may be propelled out of the bellow through the valve or the dome slit.

[1147] In certain embodiments, the reservoir compartment 1635 includes one or more valves (e.g. a valve in the dispensing outlet 1607) that are configured to move or open to fluidly couple the reservoir compartment 1635 to the GI tract. In certain embodiments, a housing wall of the housing 1601 can form a portion of the reservoir compartment 1635. In certain embodiments, the housing walls of the reservoir serve as a gasket. One or more of the one or more valves are positioned in the housing wall of the device housing 1601, in accordance with particular implementations. One or more conduits may extend from the reservoir 1635 to the one or more valves, in certain implementations.

[1148] In certain embodiments, a housing wall of the housing 1601 can be formed of a material that is configured to dissolve, for example, in response to contact at the disease site. In certain embodiments, a housing wall of the housing 1601 can be configured to dissolve in response to a chemical reaction or an electrical signal. The one or more valves and/or the signals for causing the housing wall of the housing 1601 to dissolve or dissipate can be controlled by one or more processors or controllers positioned on PCB 1632 in the device housing 1601. The controller is communicably coupled to one or more sensors or detectors configured to determine when the device housing 1601 is proximate to a disease site. The sensors or detectors comprise a plurality of electrodes comprising a coating, in certain implementations. Releasing of the immune modulator (e.g., any of the immune modulators described herein) from the reservoir compartment 1635 is triggered by an electric signal from the electrodes resulting from the interaction of the coating with the one or more sites of disease site. The one or more sensors can include a chemical sensor, an electrical sensor, an optical sensor, an electromagnetic sensor, a light sensor, a gas sensor, and/or a radiofrequency sensor. Methods for detecting volatile organic compounds (VOCs) and other gases from a biological sample include resistive metal oxide gas sensors/mixed metal oxide gas sensors, electrochemical gas sensors, optical/IR gas sensors, conducting polymer/composite polymer resistive/capacitive gas sensors, quartz crystal microbalance gas sensors, carbon nanotubes, and pellister/calorimetric gas sensors. Examples of ingestible gas sensors are described in US Patent Publication No. US20130289368, which published on Oct. 31, 2013, US Patent Publication No. US20170284956, which published on Oct. 5, 2017, and PCT Patent Publication No. WO2016197181, which published on Dec. 15, 2016. Examples of gases that can be detected in the gastrointestinal tract using a sensor include, but are not limited to, oxygen, hydrogen, and carbon dioxide.

[1149] In particular embodiments, the device housing 1601 can include one or more pumps configured to pump the therapeutically effective amount of the immune modulator from the reservoir compartment 1635. The pump is communicably coupled to the one or more controllers. The controller is configured to activate the pump in response to detection by the one or more detectors of the disease site and activation of the valves to allow the reservoir 1635 to be in fluid communication with the GI tract. The pump can include a fluid actuated pump, an electrical pump, or a mechanical pump.

[1150] In certain embodiments, the device housing 1601 comprises one or more anchor systems for anchoring the device housing 1601 or a portion thereof at a particular location in the GI tract adjacent the disease site. In some embodiments, a storage reservoir comprises an anchor system, and the storage reservoir comprising a releasable substance is anchored to the GI tract. The anchor system can be activated by the controller in response to detection by the one or more detectors of the intended site of release. In certain implementations, the anchor system includes legs or spikes configured to extend from the housing wall(s) of the device housing 1601. The spikes can be configured to retract and/or can be configured to dissolve over time. An example of an attachable device that becomes fixed to the interior surface of the GI tract is described in PCT Patent Application PCT/US2015/012209, Gastrointestinal Sensor Implantation System, filed Jan. 21, 2015, which is hereby incorporated by reference herein in its entirety.

[1151] FIG. 20 provides an example structural diagram having a flexible diaphragm 1665 that may deform towards the dispensing outlet 1607 when the gas generating cell 1603 generates gas. The dispensable substance may then be propelled by the deformed diaphragm out of the housing through the dispensing outlet 1607. The dispensing outlet 1607 shown at FIG. 20 is in the form of a ring valve, however, any outlet design can be applied.

[1152] In some embodiments, an ingestible device can have an umbrella-shaped exit valve structure as a dispensing outlet of the ingestible device. Optionally, an ingestible device can have a flexible diaphragm to deform for drug delivery, and/or an integrated piston and gas generating cell such that the gas generating cell is movable with the piston to push for drug delivery.

[1153] In certain embodiments, an ingestible device can be anchored within the intestine by extending hooks from the ingestible device after it has entered the region of interest. For example, when the ingestible device determines it has arrived at a location within the GI tract, the hooks can be actuated to extend outside of the ingestible device to catch in the intestinal wall and hold the ingestible device in the respective location. In some embodiments, the hook can pierce into the intestinal wall to hold the ingestible device 100 in place. The hooks can be hollow. A hollow hook can be used to anchor the ingestible device and/or to dispense a substance from the dispensable substance, e.g., into the intestinal wall.

[1154] In some embodiments an ingestible device includes an intestinal gripper to grip a portion of the intestinal wall for delivering the dispensable substance. Such a gripper can include two or more arms configured to out of the device and close to grip a portion of the intestinal wall.

[1155] An injecting needle can be used with the anchoring arms to inject dispensable substance into the intestinal wall after a portion of the intestinal wall is gripped.

[1156] In some embodiments, when the gas generating cell generates gas to propel the piston to move towards the nozzle such that the dispensable substance can be pushed under the pressure to break a burst disc to be injected via the nozzle.

[1157] In some embodiments, an ingestible device has a jet delivery mechanism with enhanced usable volume of dispensable substance. For example, the nozzle may be placed at the center of the ingestible device, and gas channels may be placed longitudinally along the wall of the ingestible device to transport gas from the gas generating cell to propel the piston, which is placed at an end of the ingestible device.

[1158] In some embodiments, the ingestible device can use osmotic pressure to adhere a suction device of the ingestible device to the intestinal wall. For example, the ingestible device may have an osmotic mechanism that has a chamber storing salt crystals. The chamber can include a mesh placed in proximate to a burst valve at one end of the chamber, and a reverse osmosis (RO) membrane placed in proximate to a valve on the other end of the chamber. A suction device, e.g., two or more suction fingers, is placed outside of the chamber with an open outlet exposed to luminal fluid in the GI tract. When the osmotic mechanism is inactivated, e.g., the valve is closed so that no luminal fluid is drawn into the osmotic chamber. When the osmotic mechanism is activated by opening the valve, luminal fluid enters the ingestible device through an outlet of the suction device and enters the osmotic chamber through the valve. The salt in the chamber is then dissolved into the fluid. The RO membrane prevents any fluid to flow in the reverse direction, e.g., from inside the chamber to the valve. The fluid continues to flow until all the salt contained in the chamber is dissolved or until intestinal tissue is drawn into the suction device. As luminal fluid keeps flowing into the chamber, the solution of the luminal fluid with dissolved salt in the chamber may reduce osmotic pressure such that the suction force at may also be reduced. In this way, suction of the intestinal tissue may stall before the tissue is in contact with the valve to avoid damage to the intestinal tissue.

[1159] An ingestible device employing an osmotic mechanism can also include a suction device as illustrated. The suction device can be two or more suction fingers 347a-b disposed proximate to the outlet. The outlet can be connected to a storage reservoir storing the dispensable substance (e.g., therapeutic agent). The storage reservoir can contact a piston (similar to 104 in FIG. 16), which can be propelled by pressure generated from the osmotic pump to move towards the outlet. The osmotic pump can be similar to the osmotic mechanism described in the preceding paragraph. A breakaway section can be placed in proximate to the other end (opposite to the end where the outlet 107 is disposed) of the ingestible device.

[1160] In some embodiments, tumbling suction by an ingestible device is used. Such an ingestible device does not require any electronics or other actuation elements. Such an ingestible device may constantly, intermittently, or periodically tumble when travelling through the intestine. When the ingestible device tumbles to a position that the outlet is in direct contact with the intestinal wall, a suction process similar to that described in the preceding paragraph may occur. Additional structural elements such as fins, flutes or the like may be added to the outer wall of the ingestible device 100 to promote the tumbling motion.

[1161] In certain embodiments, the reservoir is an anchorable reservoir, which is a reservoir comprising one or more anchor systems for anchoring the reservoir at a particular location in the GI tract adjacent to the intended site of delivery of the immune modulator. In certain embodiments, the anchor system includes legs or spikes or other securing means such as a piercing element, a gripping element, a magnetic-flux-guiding element, or an adhesive material, configured to extend from the anchorable reservoir of the device housing. The spikes can be configured to retract and/or can be configured to dissolve over time. In some embodiments, the anchorable reservoir is suitable for localizing, positioning and/or anchoring. In some embodiments, the anchorable reservoir is suitable for localizing, and positioning and/or anchoring by an endoscope. In some embodiments, the anchorable reservoir is connected to the endoscope. In some embodiments, the anchorable reservoir is connected to the endoscope in a manner suitable for oral administration. In some embodiments, the anchorable reservoir is connected to the endoscope in a manner suitable for rectal administration. Accordingly, provided herein in some embodiments is an anchorable reservoir is connected to an endoscope wherein the anchorable reservoir comprises a therapeutically effective amount of any of the agents described herein. In some embodiments the endoscope is fitted with a spray catheter.

[1162] Exemplary embodiments of anchorable reservoirs are as follows. In more particular examples of the following exemplary embodiments the reservoir is connected to an endoscope.

[1163] In one embodiment, the anchorable reservoir comprises an implant capsule for insertion into a body canal to apply radiation treatment to a selected portion of the body canal. The reservoir includes a body member defining at least one therapeutic treatment material receiving chamber and at least one resilient arm member associated with the body member for removably engaging the body canal when the device is positioned therein.

[1164] In one embodiment the anchorable reservoir has multiple suction ports and permits multiple folds of tissue to be captured in the suction ports with a single positioning of the device and attached together by a tissue securement mechanism such as a suture, staple or other form of tissue bonding. The suction ports may be arranged in a variety of configurations on the reservoir to best suit the desired resulting tissue orientation.

[1165] In some embodiments an anchorable reservoir comprises a tract stimulator and/or monitor IMD comprising a housing enclosing electrical stimulation and/or monitoring circuitry and a power source and an elongated flexible member extending from the housing to an active fixation mechanism adapted to be fixed into the GI tract wall is disclosed. After fixation is effected, the elongated flexible member bends into a preformed shape that presses the housing against the mucosa so that forces that would tend to dislodge the fixation mechanism are minimized. The IMD is fitted into an esophageal catheter lumen with the fixation mechanism aimed toward the catheter distal end opening whereby the bend in the flexible member is straightened. The catheter body is inserted through the esophagus into the GI tract cavity to direct the catheter distal end to the site of implantation and fix the fixation mechanism to the GI tract wall. The IMD is ejected from the lumen, and the flexible member assumes its bent configuration and lodges the hermetically sealed housing against the mucosa. A first stimulation/sense electrode is preferably an exposed conductive portion of the housing that is aligned with the bend of the flexible member so that it is pressed against the mucosa. A second stimulation/sense electrode is located at the fixation site.

[1166] In some embodiments a reservoir for sensing one or more parameters of a patient is anchored to a tissue at a specific site and is released from a device, using a single actuator operated during a single motion. As an example, a delivery device may anchor the capsule to the tissue site and release the reservoir from the delivery device during a single motion of the actuator.

[1167] In some embodiments a device is provided comprising: a reservoir configured to contain a fluid, the reservoir having at least one outlet through which the fluid may exit the reservoir; a fluid contained within the reservoir; a primary material contained within the reservoir and having a controllable effective concentration in the fluid; and at least one electromagnetically responsive control element located in the reservoir or in a wall of the reservoir and adapted for modifying the distribution of the primary material between a first active form carried in the fluid and a second form within the reservoir in response to an incident electromagnetic control signal, the effective concentration being the concentration of the first active form in the fluid, whereby fluid exiting the reservoir carries the primary material in the first active form at the effective concentration.

[1168] In some embodiments systems and methods are provided for implementing or deploying medical or veterinary devices or reservoirs (a) operable for anchoring at least partly within a digestive tract, (b) small enough to pass through the tract per vias naturales and including a wireless-control component, (c) having one or more protrusions positionable adjacent to a mucous membrane, (d) configured to facilitate redundant modes of anchoring, (e) facilitating a primary material supply deployable within a stomach for an extended and/or controllable period, (f) anchored by one or more adaptable extender modules supported by a subject's head or neck, and/or (g) configured to facilitate supporting at least a sensor within a subject's body lumen for up to a day or more.

[1169] In certain embodiments, the reservoir is attachable to an ingestible device. In certain embodiments, the ingestible device comprises a housing and the reservoir is attachable to the housing. In certain embodiments, the attachable reservoir is also an anchorable reservoir, such as an anchorable reservoir comprising one or more anchor systems for anchoring the reservoir at a particular location in the GI tract as disclosed hereinabove.

[1170] Accordingly, in certain embodiments, provided herein is an immune modulator (e.g., any of the immune modulators described herein) for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm as disclosed herein, wherein the immune modulator is contained in a reservoir suitable for attachment to a device housing, and wherein the method comprises attaching the reservoir to the device housing to form the ingestible device, prior to orally administering the ingestible device to the subject.

[1171] In certain embodiments, provided herein is an attachable reservoir containing an immune modulator (e.g., any of the immune modulators described herein) for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm, wherein the method comprises attaching the reservoir to a device housing to form an ingestible device and orally administering the ingestible device to a subject, wherein the immune modulator is released by device at a location in the gastrointestinal tract of the subject that is proximate to the intended site of release of the immune modulator.

[1172] In certain embodiments, provided herein is an attachable reservoir containing an immune modulator, wherein the reservoir is attachable to a device housing to form an ingestible device that is suitable for oral administration to a subject and that is capable of releasing the immune modulator at a location in the gastrointestinal tract of the subject that is proximate to the intended site of release.

[1173] In particular implementation the ingestible device includes cameras (e.g., video cameras) that affords inspection of the entire GI tract without discomfort or the need for sedation, thus avoiding many of the potential risks of conventional endoscopy. Video imaging can be used to help determine one or more characteristics of the GI tract. In some embodiments, the ingestible device 101 may comprise a camera for generating video imaging data of the GI tract which can be used to determine, among other things, the location of the device. Examples of video imaging capsules include Medtronic's PillCam, Olympus' Endocapsule, and IntroMedic's MicroCam. For a review of imaging capsules, see Basar et al. Ingestible Wireless Capsule Technology: A Review of Development and Future Indication International Journal of Antennas and Propagation (2012); 1-14). Other imaging technologies implemented with the device 101 can include thermal imaging cameras, and those that employ ultrasound or Doppler principles to generate different images (see Chinese patent application CN104473611: Capsule endoscope system having ultrasonic positioning function.

[1174] Ingestible devices can be equipped with sources for generating reflected light, including light in the Ultraviolet, Visible, Near-infrared and/or Mid-infrared spectrum, and the corresponding detectors for spectroscopy and hyperspectral imaging. Likewise, autofluorescense may be used to characterize GI tissue (e.g., subsurface vessel information), or low-dose radiation (see Check-Cap) can be used to obtain 3D reconstructed images.

Device Components

[1175] An ingestible device in accordance with particular embodiments of the present invention may comprise a component made of a non-digestible material and contain the immune modulator (e.g., any of the immune modulators described herein). In some embodiments, the material is plastic.

[1176] It is envisaged that the device is single-use. The device is loaded with a drug prior to the time of administration. In some embodiments, it may be preferred that there is provided a medicinal product comprising the device pre-filled with the drug.

Anchoring Components

[1177] Several systems may actively actuate and control the capsule position and orientation in different sections of the GI tract. Examples include leg-like or anchor-like mechanisms that can be deployed by an ingestible device to resist peristaltic forces in narrowed sections of the GI tract, such as the intestine, and anchor the device to a location. Other systems employ magnetic shields of different shapes that can interact with external magnetic fields to move the device. These mechanisms may be particularly useful in areas outside of the small intestine, like the cecum and large intestine.

[1178] An anchoring mechanism may be a mechanical mechanism. For example, a device may be a capsule comprising a plurality of legs configured to steer the capsule. The number of legs in the capsule may be, for example, two, four, six, eight, ten or twelve. The aperture between the legs of the device may be up to about 35 mm; about 30 to about 35 mm; about 35 to about 75 mm; or about 70 to about 75 mm. The contact area of each leg may be varied to reduce impact on the tissue. One or more motors in the capsule may each actuate a set of legs independently from the other. The motors may be battery-powered motors.

[1179] An anchoring mechanism may be a non-mechanical mechanism. For example, a device may be a capsule comprising a permanent magnet located inside the capsule. The capsule may be anchored at the desired location of the GI tract by an external magnetic field.

[1180] An anchoring mechanism may comprise a non-mechanical mechanism and a mechanical mechanism. For example, a device may be a capsule comprising one or more legs, one or more of which are coated with an adhesive material.

Locomotion Components

[1181] Ingestible devices can be active or passive, depending on whether they have controlled or non-controlled locomotion. Passive (non-controlled) locomotion is more commonly used among ingestible devices given the challenges of implementing a locomotion module. Active (controlled) locomotion is more common in endoscopic ingestible capsules. For example, a capsule may comprise a miniaturized locomotion system (internal locomotion). Internal locomotion mechanisms may employ independent miniaturized propellers actuated by DC brushed motors, or the use of water jets. As an example, a mechanism may comprise flagellar or flap-based swimming mechanisms. As an example, a mechanism may comprise cyclic compression/extension shape-memory alloy (SMA) spring actuators and anchoring systems based on directional micro-needles. As an example, a mechanism may comprise six SMA actuated units, each provided with two SMA actuators for enabling bidirectional motion. As an example, a mechanism may comprise a motor adapted to electrically stimulating the GI muscles to generate a temporary restriction in the bowel.

[1182] As an example, a capsule may comprise a magnet and motion of the capsule is caused by an external magnetic field. For example, a locomotion system may comprise an ingestible capsule and an external magnetic field source. For example, the system may comprise an ingestible capsule and magnetic guidance equipment such as, for example, magnetic resonance imaging and computer tomography, coupled to a dedicated control interface.

[1183] In some embodiments drug release mechanisms may also be triggered by an external condition, such as temperature, pH, movement, acoustics, or combinations thereof.

Use of an Endoscope or an Ingestible Device in Biopsy and Surgery

Sampling

[1184] Ingestible devices may comprise a mechanism adapted to permit the collection of tissue samples. In some examples, this is achieved using electro-mechanical solutions to collect and store the sample inside an ingestible device. As an example, a biopsy mechanism may include a rotational tissue cutting razor fixed to a torsional spring or the use of microgrippers to fold and collect small biopsies. As an example, Over-the-scope clips (OTSC) may be used to perform endoscopic surgery and/or biopsy. As an example of the methods disclosed herein, the method may comprise releasing an immune modulator (e.g., any of the immune modulators described herein) and collecting a sample inside the device. As an example, the method may comprise releasing an immune modulator and collecting a sample inside the device in a single procedure.

[1185] FIG. 21 illustrates an example ingestible device 2100 with multiple openings in the housing. The ingestible device 2100 has an outer housing with a first end 2102A, a second end 2102B, and a wall 2104 extending longitudinally from the first end 2102A to the second end 2102B. Ingestible device 2100 has a first opening 2106 in the housing, which is connected to a second opening 2108 in the housing. The first opening 2106 of the ingestible device 2100 is oriented substantially perpendicular to the second opening 2108, and the connection between the first opening 2106 and the second opening 2108 forms a curved chamber 2110 within the ingestible device 2100.

[1186] The overall shape of the ingestible device 2100, or any of the other ingestible devices discussed in this disclosure, may be similar to an elongated pill or capsule.

[1187] In some embodiments, a portion of the curved chamber 2110 may be used as a sampling chamber, which may hold samples obtained from the GI tract. In some embodiments the curved chamber 2110 is subdivided into sub-chambers, each of which may be separated by a series of one or more valves or interlocks.

[1188] In some embodiments, the first opening 2106, the second opening 2108, or the curved chamber 2110 include one or more of a hydrophilic or hydrophobic material, a sponge, a valve, or an air permeable membrane.

[1189] The use of a hydrophilic material or sponge may allow samples to be retained within the curved chamber 2110, and may reduce the amount of pressure needed for fluid to enter through the first opening 2106 and dislodge air or gas in the curved chamber 2110. Examples of hydrophilic materials that may be incorporated into the ingestible device 2100 include hydrophilic polymers such as polyvinyl alcohol, polyvinyl pyrrolidone, and the like. Similarly, materials that have undergone various types of treatments, such as plasma treatments, may have suitable hydrophilic properties, and may be incorporated into the investible device 2100. Sponges may be made of any suitable material or combination of materials, such as fibers of cotton, rayon, glass, polyester, polyethylene, polyurethane, and the like. Sponges generally may be made from commercially available materials, such as those produced by Porex.

[1190] As discussed in more detail below, in some embodiments, the sponges may be treated in order to change their absorbency or to help preserve samples.

[1191] In some embodiments, the sponges may be cut or abraded to change their absorbency or other physical properties.

[1192] Hydrophobic materials located near the second opening 2108 may repel liquids, discouraging liquid samples from entering or exiting the curved chamber 2110 through the second opening 2108. This may serve a similar function as an air permeable membrane. Examples of hydrophobic materials which may be incorporated into the ingestible device 2100 include polycarbonate, acrylics, fluorocarbons, styrenes, certain forms of vinyl, and the like.

[1193] The various materials listed above are provided as examples, and are not limiting. In practice, any type of suitable hydrophilic, hydrophobic, or sample preserving material may be used in the ingestible device 2100.

[1194] In some embodiments, an ingestible device includes a moveable valve as a diaphragm valve, which uses a mechanical actuator to move a flexible diaphragm in order to seal or unseal an aperture in a second portion of an inlet region, which may effectively block or unblock the inlet region. However, it will be understood that, in some embodiments, the moveable valve may be a different type of valve. For example, in some embodiments the moveable valve may be replaced by a pumping mechanism. As another example, in some embodiments the moveable valve is replaced with an osmotic valve

[1195] A sampling chamber of an ingestible device can have an exit port to allow air or gas to exit the sampling chamber, while preventing at least a portion of the sample obtained by the ingestible device from exiting the sampling chamber. For example, the exit port may include a gas-permeable membrane. An ingestible device can include one-way valve as part of its exit port.

[1196] An ingestible device can include an outlet port connected to the volume within housing of the ingestible device. The outlet port may provide a path for the gas to exit the ingestible device and be released into the environment surrounding the ingestible device. This may prevent pressure from building up within the housing of the ingestible device. In some embodiments, an ingestible device does not include an outlet port, and the gas stays inside the volume of the ingestible device. In some embodiments, the outlet port may contain a gas permeable membrane, a one-way valve, a hydrophobic channel, or some other mechanism to avoid unwanted material, (e.g., fluids and solid particulates from within the GI tract), from entering the ingestible device through the outlet port.

[1197] In some embodiments, the ingestible device may include a sensor within or proximate to the sampling chamber. For example, this sensor may be used to detect various properties of a sample contained within the sampling chamber, or this sensor may be used to detect the results of an assay technique applied to the sample contained within the sampling chamber.

[1198] In some embodiments, a hydrophilic sponge is located within the sampling chamber, and the hydrophilic sponge may be configured to absorb the sample as the sample enters the sampling chamber. In some embodiments, the hydrophilic sponge fills a substantial portion of the sampling chamber, and holds the sample for an extended period of time. This may be particularly advantageous if the sample is collected from the ingestible device after the ingestible device exits the body. In some embodiments, the hydrophilic sponge is placed on only certain surfaces or fills only certain portions of the sampling chamber. For example, it may be possible to line certain walls (or all walls) of the sampling chamber with a hydrophilic sponge to assist in drawing in the sample, while leaving some (or none) of the walls of the sampling chamber uncovered. Leaving walls uncovered may allow the use of diagnostics or assay techniques that require a relatively un-obscured optical path.

[1199] In some embodiments, the ingestible device may include a sealed vacuum chamber connected to the exit port, or connected directly or indirectly to the sampling chamber. In some embodiments a pin valve may be used as a moveable valve (e.g., as moveable valve of ingestible device). In certain embodiments, a rotary valve may be used as a moveable valve (e.g., as moveable valve of ingestible device). In some embodiments, a flexible diaphragm, or diaphragm valve, may be used as a moveable valve (e.g., as moveable valve of ingestible device). In certain embodiments, a mechanism is near the diaphragm or in direct contact with the diaphragm. The spring mechanism may apply pressure to the diaphragm to oppose the pressure applied by the mechanical actuator, which may cause the flexible diaphragm to be moved into an open position when the mechanical actuator is not applying pressure to the flexible diaphragm. Additionally, this may ensure that the diaphragm valve remains open when the mechanical actuator is not applying pressure across the flexible diaphragm. In some embodiments, moving the mechanical actuator from a closed position to an open position causes a volume of the inlet region within the ingestible device to increase. This may cause the pressure within the inlet region to be reduced, generating suction to draw a sample into the inlet region. Similarly, moving the mechanical actuator from an open position to a closed position may cause the volume of the inlet region to be reduced. This may cause the pressure within the inlet region to be increased, pushing the sample out of the inlet region. Depending on the design of the inlet region, the mechanical actuator, and the moveable valve, this may push the sample into the sampling chamber rather than pushing the sample back through the opening in the ingestible device.

[1200] FIG. 22 depicts a cross-sectional view of a portion of the interior of ingestible device 3000. As shown in FIG. 22, the interior of ingestible device 3000 includes a valve system 3100 and a sampling system 3200. Valve system 3100 is depicted as having a portion that is flush with the opening 3018 so that valve system 3100 prevents fluid exterior to ingestible device 2000 from entering sampling system 3200. However, as described in more detail below with reference to FIGS. 22-27, valve system 3100 can change position so that valve system 3100 allows fluid exterior to ingestible device 3000 to enter sampling system 3200.

[1201] FIGS. 23 and 27 illustrate valve system 3100 in more detail. As shown in FIG. 23, valve system 3100 includes an actuation mechanism 3110, a trigger 3120, and a gate 3130. In FIGS. 23 and 7, a leg 3132 of gate 3130 is flush against, and parallel with, housing wall 3016 so that gate leg 3132 covers opening 3018 to prevent fluid exterior to ingestible device 3000 (e.g., fluid in the GI tract) from entering the interior of ingestible device 3000. A protrusion 3134 of gate 3130 engages a lip 3122 of trigger 3120. A peg 3124 of trigger 3120 engages a wax pot 3112 of actuation mechanism 3110. Referring to FIG. 27, a biasing mechanism 3140 includes a compression spring 3142 that applies an upward force on gate 3130. Biasing mechanism 3140 also includes a torsion spring 3144 that applies a force on trigger 3120 in the counter-clockwise direction. In FIGS. 23 and 27, the force applied by torsion spring 3144 is counter-acted by the solid wax in pot 3112, and the force applied by compression spring 3142 is counter-acted by lip 3122.

[1202] FIG. 24A and FIG. 24B show an embodiment of the manner in which actuation mechanism 3110 actuates movement of trigger 3120. Similar to FIGS. 23 and 27, FIG. 24A shows a configuration in which peg 3124 applies a force against solid wax pot 3112 due to torsion spring 3144, and in which the solid nature of wax pot 3112 resists the force applied by peg 3124. A control unit 3150 is in signal communication with valve system 3100. During use of ingestible device 3000, a control unit 3150 receives a signal, indicating that the position of valve system 3100 should change, e.g., so that ingestible device 3000 can take a sample of a fluid in the GI tract. Control unit 3150 sends a signal that causes a heating system 3114 of actuation system 3100 to heat the wax in pot 3112 so that the wax melts. As shown in FIG. 24B, the melted wax is not able to resist the force applied by peg 3124 so that, under the force of torsion spring 3144, trigger 3120 moves in a counter-clockwise fashion.

[1203] FIGS. 25A and 25B illustrate the interaction of trigger 3120 and gate 3130 before and after actuation. As shown in FIG. 25A, when wax pot 3112 is solid (corresponding to the configuration shown in FIG. 24A), protrusion 3134 engages lip 3122, which prevents the force of compression spring 3142 from moving gate 3130 upward. As shown in FIG. 25B, when the wax in pot 3112 melts (FIG. 24B), trigger 3120 moves counter-clockwise, and lip 3122 disengages from protrusion 3134. This allows the force of compression spring 3142 to move gate 3130 upward. As seen by comparing FIG. 25A to FIG. 25B, the upward movement of gate 3130 results in an upward movement of an opening 3136 in gate leg 3132.

[1204] FIGS. 26A and 26B illustrate the impact of the upward movement of opening 3136 on the ability of ingestible device 3000 to obtain a sample. As shown in FIG. 26A, when the wax in pot 3112 is solid (FIGS. 24A and 25A), opening 3136 in is not aligned with opening 3018 in wall 3016 of ingestible device 3000. Instead, gate leg 3132 covers opening 3018 and blocks fluid from entering the interior of ingestible device 3000. As shown in FIG. 26B, when the wax in pot 3112 is melted and trigger 3120 and gate 3130 have moved (FIGS. 24B and 42B), opening 3136 in gate 3130 is aligned with opening 3018 in wall 3016. In this configuration, fluid that is exterior to ingestible device 3000 (e.g., in the GI tract) can enter the interior of ingestible device 3000 via openings 3018 and 3036.

[1205] FIG. 27 illustrates a more detailed view of ingestible device 3000 including valve system 3100 and sampling system 3200.

[1206] While the foregoing description is made with regard to a valve system having one open position and one closed position (e.g., a two-stage valve system), the disclosure is not limited in this sense. Rather, the concepts described above with regard to a two stage valve system can be implemented with a valve system have more than two stages (e.g., three stages, four stages, five stages, etc.).

[1207] As noted above in addition to a valve system, an ingestible device includes a sampling system. FIG. 28 illustrates a partial cross sectional view of ingestible device 3000 with sampling system 3200 and certain components of valve system 3100. Sampling system 3200 includes a series of sponges configured to absorb fluid from an opening, move the fluid to a location within the housing, and prepare the fluid for testing. Preparation for testing may include filtering the fluid and combining the fluid with a chemical assay. The assay may be configured to dye cells in the filtered sample. The series of sponges includes a wicking sponge 3210, a transfer sponge 3220, a volume sponge 3230, and an assay sponge 3240. Sampling system 3200 also includes a membrane 3270 located between assay sponge 3240 and a vent 3280 for gases to leave sampling system 3200. A cell filter 3250 is located between distal end 3214 of wicking sponge 3210 and a first end 3222 of transfer sponge 3220. Membrane 3270 is configured to allow one or more gases to leave sampling system 3200 via an opening 3280, while maintaining liquid in sampling system 3200.

[1208] FIG. 29 is a highly schematic illustration of an ingestible device 4000 that contains multiple different systems that cooperate for obtaining a sample and analyzing a sample, e.g., within the GI tract of a subject. Ingestible device 4000 includes a power system 4100 (e.g., one or more batteries), configured to power an electronics system 4200 (e.g., including a control system, optionally in signal communication with an external base station), a valve system 4300, a sampling system 4400, and an analytic system 4500. Exemplary analytical systems include assay systems, such as, for example, optical systems containing one or more sources of radiation and/or one more detectors.

[1209] Some or all of the sponges of the above-described sampling systems may contain one or more preservatives (see discussion above). Typically, the assay sponge and/or the volume sponge 3230 and/or the transfer sponge contain one or more preservatives. Typically, the preservative(s) are selected based on the analyte of interest, e.g., an analyte (such as a protein biomarker) for a GI disorder.

Communication Systems

[1210] An ingestible device may be equipped with a communication system adapted to transmit and/or receive data, including imaging and/or localization data. As an example, a communication system may employ radiofrequency transmission. Ingestible devices using radiofrequency communication are attractive because of their efficient transmission through the layers of the skin. This is especially true for low frequency transmission (UHF-433 ISM and lower, including the Medical Device Radio Communication Service band (MDRS) band 402-406 MHz). In another embodiment, acoustics are used for communications, including the transmission of data. For example, an ingestible capsule may be able to transmit information by applying one or more base voltages to an electromechanical transducer or piezoelectric (e.g., PZT, PVDF, etc.) device to cause the piezoelectric device to ring at particular frequencies, resulting in an acoustic transmission. A multi-sensor array for receiving the acoustic transmission may include a plurality of acoustic transducers that receive the acoustic transmission from a movable device such as an ingestible capsule as described in U.S. patent application Ser. No. 11/851,214 filed Sep. 6, 2007, incorporated by reference herein in its entirety.

[1211] As an example, a communication system may employ human body communication technology. Human body communication technology uses the human body as a conductive medium, which generally requires a large number of sensor electrodes on the skin. As an example, a communication system may integrate a data storage system.

Environmental Sensors

[1212] In some embodiments the device may comprise environmental sensors to measure pH, temperature, transit times, or combinations thereof. Other examples of environmental sensors include, but are not limited to a capacitance sensor, an impedance sensor, a heart rate sensor, acoustic sensor such as a microphone or hydrophone, image sensor, and/or a movement sensor. In one embodiment, the ingestible device comprises a plurality of different environmental sensors for generating different kinds of environmental data.

[1213] In order to avoid the problem of capsule retention, a thorough past medical and surgical history should be undertaken. In addition, several other steps have been proposed, including performing investigations such as barium follow-through. In cases where it is suspected that there is a high risk of retention, the patient is given a patency capsule a few days before swallowing an ingestible device. Any dissolvable non-endoscopic capsule may be used to determine the patency of the GI tract. The patency capsule is usually the same size as the ingestible device and can be made of cellophane. In some embodiments, the patency capsule contains a mixture of barium and lactose, which allows visualization by x-ray. The patency capsule may also include a radiotag or other label, which allows for it to be detected by radio-scanner externally. The patency capsule may comprise wax plugs, which allow for intestinal fluid to enter and dissolve the content, thereby dividing the capsule into small particles.

[1214] Accordingly, in some embodiments, the methods herein comprise (a) identifying a subject having an inflammatory disease or condition that arises in a tissue originating from the endoderm and (b) evaluating the subject for suitability to treatment. In some embodiments, the methods herein comprise evaluating for suitability to treatment a subject identified as having a disease or condition that arises in a tissue originating from the endoderm. In some embodiments, evaluating the subject for suitability to treatment comprises determining the patency of the subject's GI tract.

[1215] In some embodiments, an ingestible device comprises a tissue anchoring mechanism for anchoring the ingestible device to a subject's tissue. For example, an ingestible device could be administered to a subject and once it reaches the desired location for release of the immune modulator (e.g., any of the immune modulators described herein), the tissue attachment mechanism can be activated or deployed such that the ingestible device, or a portion thereof, is anchored to the desired location. In some embodiments, the tissue anchoring mechanism is reversible such that after initial anchoring, the tissue attachment device is retracted, dissolved, detached, inactivated or otherwise rendered incapable of anchoring the ingestible device to the subject's tissue. In some embodiments the attachment mechanism is placed endoscopically.

[1216] In some embodiments, a tissue anchoring mechanism comprises an osmotically-driven sucker. In some embodiments, the osmotically-driven sucker comprises a first valve on the near side of the osmotically-driven sucker (e.g., near the subject's tissue) and a second one-way valve that is opened by osmotic pressure on the far side of the osmotically-driven sucker, and an internal osmotic pump system comprising salt crystals and semi-permeable membranes positioned between the two valves. In such embodiments, osmotic pressure is used to adhere the ingestible device to the subject's tissue without generating a vacuum within the ingestible capsule. After the osmotic system is activated by opening the first valve, fluid is drawn in through the sucker and expelled through the second burst valve. Fluid continues to flow until all the salt contained in the sucker is dissolved or until tissue is drawn into the sucker. As liminal fluid is drawn through the osmotic pump system, solutes build up between the tissue and the first valve, reducing osmotic pressure. In some embodiments, the solute buildup stalls the pump before the tissue contacts the valve, preventing tissue damage. In some embodiments, a burst valve is used on the far side of the osmotically-driven sucker rather than a one-way valve, such that luminal fluid eventually clears the saline chamber and the osmotic flow reverses, actively pushing the subject's tissue out of the sucker. In some embodiments, the ingestible device may be anchored to the interior surface of tissues forming the GI tract of a subject. In one embodiment, the ingestible device comprises a connector for anchoring the device to the interior surface of the GI tract. The connector may be operable to ingestible device to the interior surface of the GI tract using an adhesive, negative pressure and/or fastener.

[1217] In some embodiments a device comprises a tract stimulator and/or monitor IMD comprising a housing enclosing electrical stimulation and/or monitoring circuitry and a power source and an elongated flexible member extending from the housing to an active fixation mechanism adapted to be fixed into the GI tract wall is disclosed. After fixation is effected, the elongated flexible member bends into a preformed shape that presses the housing against the mucosa so that forces that would tend to dislodge the fixation mechanism are minimized. The IMD is fitted into an esophageal catheter lumen with the fixation mechanism aimed toward the catheter distal end opening whereby the bend in the flexible member is straightened. The catheter body is inserted through the esophagus into the GI tract cavity to direct the catheter distal end to the site of implantation and fix the fixation mechanism to the GI tract wall. The IMD is ejected from the lumen, and the flexible member assumes its bent configuration and lodges the hermetically sealed housing against the mucosa. A first stimulation/sense electrode is preferably an exposed conductive portion of the housing that is aligned with the bend of the flexible member so that it is pressed against the mucosa. A second stimulation/sense electrode is located at the fixation site.

[1218] In some embodiments a device includes a fixation mechanism to anchor the device to tissue within a body lumen, and a mechanism to permit selective de-anchoring of the device from the tissue anchoring site without the need for endoscopic or surgical intervention. An electromagnetic device may be provided to mechanically actuate the de-anchoring mechanism. Alternatively, a fuse link may be electrically blown to de-anchor the device. As a further alternative, a rapidly degradable bonding agent may be exposed to a degradation agent to de-anchor the device from a bonding surface within the body lumen.

[1219] In some embodiments a device is as disclosed in patent publication WO2015112575A1, incorporated by reference herein in its entirety. The patent publication is directed to a gastrointestinal sensor implantation system. In some embodiments an orally-administrable capsule comprises a tissue capture device or reservoir removably coupled to the orally-administrable capsule, where the tissue capture device including a plurality of fasteners for anchoring the tissue capture device to gastrointestinal tissue within a body

[1220] In some embodiments, the ingestible device contains an electric energy emitting means, a radio signal transmitting means, a medicament storage means and a remote actuatable medicament releasing means. The capsule signals a remote receiver as it progresses through the alimentary tract in a previously mapped route and upon reaching a specified site is remotely triggered to release a dosage of medicament. Accordingly, in some embodiments, releasing the agentis triggered by a remote electromagnetic signal.

[1221] In some embodiments, the ingestible device includes a housing introducible into a body cavity and of a material insoluble in the body cavity fluids, but formed with an opening covered by a material which is soluble in body cavity fluids. A diaphragm divides the interior of the housing into a medication chamber including the opening, and a control chamber. An electrolytic cell in the control chamber generates a gas when electrical current is passed therethrough to deliver medication from the medication chamber through the opening into the body cavity at a rate controlled by the electrical current. Accordingly, in some embodiments, releasing the immune modulator is triggered by generation in the composition of a gas in an amount sufficient to expel the immune modulator.

[1222] In some embodiments, the ingestible device includes an oral drug delivery device having a housing with walls of water permeable material and having at least two chambers separated by a displaceable membrane. The first chamber receives drug and has an orifice through which the drug is expelled under pressure. The second chamber contains at least one of two spaced apart electrodes forming part of an electric circuit which is closed by the ingress of an aqueous ionic solution into the second chamber. When current flows through the circuit, gas is generated and acts on the displaceable membrane to compress the first chamber and expel the active ingredient through the orifice for progressive delivery to the gastrointestinal tract.

[1223] In some embodiments, the ingestible device includes an ingestible device for delivering a substance to a chosen location in the GI tract of a mammal includes a receiver of electromagnetic radiation for powering an openable part of the device to an opened position for dispensing of the substance. The receiver includes a coiled wire that couples the energy field, the wire having an air or ferrite core. In a further embodiment the invention includes an apparatus for generating the electromagnetic radiation, the apparatus including one or more pairs of field coils supported in a housing. The device optionally includes a latch defined by a heating resistor and a fusible restraint. The device may also include a flexible member that may serve one or both the functions of activating a transmitter circuit to indicate dispensing of the substance; and restraining of a piston used for expelling the substance.

[1224] In some embodiments, the ingestible device includes an ingestible device for delivering a substance to a chosen location in the GI tract of a mammal includes a receiver of electromagnetic radiation for powering an openable part of the device to an opened position for dispensing of the substance. The receiver includes a coiled wire that couples the energy field, the wire having an air or ferrite core. In a further embodiment the invention includes an apparatus for generating the electromagnetic radiation, the apparatus including one or more pairs of field coils supported in a housing. The device optionally includes a latch defined by a heating resistor and a fusible restraint. The device may also include a flexible member that may serve one or both the functions of activating a transmitter circuit to indicate dispensing of the substance; and restraining of a piston used for expelling the substance.

[1225] In some embodiments, the ingestible device is a device a swallowable capsule. A sensing module is disposed in the capsule. A bioactive substance dispenser is disposed in the capsule. A memory and logic component is disposed in the capsule and in communication with the sensing module and the dispenser.

[1226] In some embodiments, localized administration is implemented via an electronic probe which is introduced into the intestinal tract of a living organism and which operates autonomously therein, adapted to deliver one or more therapy agents. In one embodiment, the method includes loading the probe with one or more therapy agents, and selectively releasing the agents from the probe at a desired location of the intestinal tract in order to provide increased efficacy over traditional oral ingestion or intravenous introduction of the agent(s).

[1227] In some embodiments, the ingestible device includes electronic control means for dispensing the drug substantially to the intended site in the GI tract, according to a pre-determined drug release profile obtained prior to administration from the specific mammal. Accordingly, in some embodiments, releasing the immune modulator (e.g., any of the immune modulators described herein) is triggered by an electromagnetic signal generated within the device. The releasing may occur according to a pre-determined drug release profile.

[1228] In some embodiments, the ingestible device can include at least one guide tube, one or more tissue penetrating members positioned in the guide tube, a delivery member, an actuating mechanism and a release element. The release element degrades upon exposure to various conditions in the intestine so as to release and actuate the actuating mechanism. Embodiments of the invention are particularly useful for the delivery of drugs which are poorly absorbed, tolerated and/or degraded within the GI tract.

[1229] In some embodiments, the ingestible device includes an electronic pill comprising at least one reservoir with a solid powder or granulate medicament or formulation, a discharge opening and an actuator responsive to control circuitry for displacing medicine from the reservoir to the discharge opening. The medicament or formulation comprises a dispersion of one or more active ingredientse.g., solids in powder or granulate formin an inert carrier matrix. Optionally, the active ingredients are dispersed using intestinal moisture absorbed into the pill via a semi-permeable wall section.

[1230] In some embodiments, the ingestible device includes a sensor comprising a plurality of electrodes having a miniature size and a lower power consumption and a coating exterior to the electrodes, wherein the coating interacts with a target condition thereby producing a change in an electrical property of the electrodes, wherein the change is transduced into an electrical signal by the electrodes. Accordingly, in some embodiments, releasing the immune modulators is triggered by an electric signal by the electrodes resulting from the interaction of the coating with the intended site of release. Further provided herein is a system for medication delivery comprising such sensor and a pill.

[1231] In some embodiments, the ingestible device includes an electronic pill comprising a plurality of reservoirs, each of the reservoirs comprising a discharge opening covered by a removable cover. The pill comprises at least one actuator responsive to control circuitry for removing the cover from the discharge opening. The actuator can for example be a spring loaded piston breaking a foil cover when dispensing the medicament. Alternatively, the cover can be a rotatable disk or cylinder with an opening which can be brought in line with the discharge opening of a reservoir under the action of the actuator.

[1232] In some embodiments, the ingestible device includes an electronically and remotely controlled pill or medicament delivery system. The pill includes a housing; a reservoir for storing a medicament; an electronically controlled release valve or hatch for dispensing one or more medicaments stored in the reservoir while traversing the gastrointestinal tract; control and timing circuitry for opening and closing the valve; and a battery. The control and timing circuitry opens and closes the valve throughout a dispensing time period in accordance with a preset dispensing timing pattern which is programmed within the control and timing circuitry. RF communication circuitry receives control signals for remotely overriding the preset dispensing timing pattern, reprogramming the control and timing circuitry or terminating the dispensing of the medicament within the body. The pill includes an RFID tag for tracking, identification, inventory and other purposes.

[1233] In some embodiments, the ingestible device includes an electronic capsule which has a discrete drive element comprising: a housing, electronics for making the electronic capsule operable, a pumping mechanism for dosing and displacing a substance, a power source for powering the electronic capsule and enabling the electronics and the pumping mechanism to operate, and a locking mechanism; and a discrete payload element comprising: a housing, a reservoir for storing the substance, one or more openings in the housing for releasing the substance from the reservoir and a locking mechanism for engaging the drive element locking mechanism. Engagement of the drive element locking mechanism with the payload element locking mechanism secures the drive element to the payload element, thereby making the electronic capsule operable and specific.

[1234] In some embodiments, the ingestible device may be a mucoadhesive device configured for release of an active agent.

[1235] In some embodiments, the ingestible device includes an apparatus that includes an ingestible medical treatment device, which is configured to initially assume a contracted state having a volume of less than 4 cm.sup.3. The device includes a gastric anchor, which initially assumes a contracted size, and which is configured to, upon coming in contact with a liquid, expand sufficiently to prevent passage of the anchor through a round opening having a diameter of between 1 cm and 3 cm. The device also includes a duodenal unit, which is configured to pass through the opening, and which is coupled to the gastric anchor such that the duodenal unit is held between 1 cm and 20 cm from the gastric anchor.

[1236] In some embodiments, the ingestible device includes a medical robotic system and method of operating such comprises taking intraoperative external image data of a patient anatomy, and using that image data to generate a modeling adjustment for a control system of the medical robotic system (e.g., updating anatomic model and/or refining instrument registration), and/or adjust a procedure control aspect (e.g., regulating substance or therapy delivery, improving targeting, and/or tracking performance).

[1237] In one embodiment the ingestible device may also include one or more environmental sensors. Environmental sensor may be used to generate environmental data for the environment external to device in the gastrointestinal (GI) tract of the subject. In some embodiments, environmental data is generated at or near the location within the GI tract of the subject where a drug is delivered. Examples of environmental sensor include, but are not limited to a capacitance sensor, a temperature sensor, an impedance sensor, a pH sensor, a heart rate sensor, acoustic sensor, image sensor (e.g., a hydrophone), and/or a movement sensor (e.g., an accelerometer). In one embodiment, the ingestible device comprises a plurality of different environmental sensors for generating different kinds of environmental data.

[1238] In one embodiment, the image sensor is a video camera suitable for obtaining images in vivo of the tissues forming the GI tract of the subject. In one embodiment, the environmental data is used to help determine one or more characteristics of the GI tract, including the location of disease (e.g., presence or location of inflamed tissue and/or lesions associated with inflammatory bowel disease). In some embodiments, the ingestible device may comprise a camera for generating video imaging data of the GI tract which can be used to determine, among other things, the location of the device.

[1239] In another embodiment, the ingestible device described herein may be localized using a gamma scintigraphy technique or other radio-tracker technology as employed by Phaeton Research's Enterion capsule (See Teng, Renli, and Juan Maya. Absolute bioavailability and regional absorption of ticagrelor in healthy volunteers. Journal of Drug Assessment 3.1 (2014): 43-50), or monitoring the magnetic field strength of permanent magnet in the ingestible device (see T. D. Than, et al., A review of localization systems for robotic endoscopic capsules, IEEE Trans. Biomed. Eng., vol. 59, no. 9, pp. 2387-2399, September 2012).

[1240] In one embodiment, the one or more environmental sensors measure pH, temperature, transit times, or combinations thereof.

[1241] In some embodiments, releasing the immune modulator (e.g., any of the immune modulators described herein) is dependent on the pH at or in the vicinity of the location. In some embodiments the pH in the jejunum is from 6.1 to 7.2, such as 6.6. In some embodiments the pH in the mid small bowel is from 7.0 to 7.8, such as 7.4. In some embodiments the pH in the ileum is from 7.0 to 8.0, such as 7.5. In some embodiments the pH in the right colon is from 5.7 to 7.0, such as 6.4. In some embodiments the pH in the mid colon is from 5.7 to 7.4, such as 6.6. In some embodiments the pH in the left colon is from 6.3 to 7.7, such as 7.0. In some embodiments, the gastric pH in fasting subjects is from about 1.1 to 2.1, such as from 1.4 to 2.1, such as from 1.1 to 1.6, such as from 1.4 to 1.6. In some embodiments, the gastric pH in fed subjects is from 3.9 to 7.0, such as from 3.9 to 6.7, such as from 3.9 to 6.4, such as from 3.9 to 5.8, such as from 3.9 to 5.5, such as from 3.9 to 5.4, such as from 4.3 to 7.0, such as from 4.3 to 6.7, such as from 4.3 to 6.4, such as from 4.3 to 5.8, such as from 4.3 to 5.5, such as from 4.3 to 5.4. In some embodiments, the pH in the duodenum is from 5.8 to 6.8, such as from 6.0 to 6.8, such as from 6.1 to 6.8, such as from 6.2 to 6.8, such as from 5.8 to 6.7, such as from 6.0 to 6.7, such as from 6.1 to 6.7, such as from 6.2 to 6.7, such as from 5.8 to 6.6, such as from 6.0 to 6.6, such as from 6.1 to 6.6, such as from 6.2 to 6.6, such as from 5.8 to 6.5, such as from 6.0 to 6.5, such as from 6.1 to 6.5, such as from 6.2 to 6.5.

[1242] In some embodiments, releasing the immune modulator (e.g., any of the immune modulators described herein) is not dependent on the pH at or in the vicinity of the location. In some embodiments, releasing the immune modulator (e.g., any of the immune modulators described herein) is triggered by degradation of a release component located in the capsule. In some embodiments, the release of the immune modulator is not triggered by degradation of a release component located in the capsule. In some embodiments, the release of the immune modulator is not dependent on enzymatic activity at or in the vicinity of the location. In some embodiments, releasing the immune modulator is not dependent on bacterial activity at or in the vicinity of the location.

[1243] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1244] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1245] a reservoir located within the housing and containing the immune modulator (e.g., any of the immune modulators described herein),

[1246] wherein a first end of the reservoir is attached to the first end of the housing;

[1247] a mechanism for releasing the immune modulator from the reservoir;

[1248] and;

[1249] an exit valve configured to allow the immune modulator to be released out of the housing from the reservoir.

[1250] In some embodiments, the ingestible device further comprises:

[1251] an electronic component located within the housing; and

[1252] a gas generating cell located within the housing and adjacent to the electronic component,

[1253] wherein the electronic component is configured to activate the gas generating cell to generate gas.

[1254] In some embodiments, the ingestible device further comprises:

a safety device placed within or attached to the housing,

[1255] wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

[1256] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1257] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1258] an electronic component located within the housing;

[1259] a gas generating cell located within the housing and adjacent to the electronic component, [1260] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1261] a reservoir located within the housing, [1262] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1263] an exit valve located at the first end of the housing, [1264] wherein the exit valve is configured to allow the dispensable substance to be released out of the first end of the housing from the reservoir; and

[1265] a safety device placed within or attached to the housing, [1266] wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

[1267] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1268] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1269] an electronic component located within the housing,

[1270] a gas generating cell located within the housing and adjacent to the electronic component, [1271] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1272] a reservoir located within the housing, [1273] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1274] an injection device located at the first end of the housing, [1275] wherein the jet injection device is configured to inject the dispensable substance out of the housing from the reservoir; and

[1276] a safety device placed within or attached to the housing, [1277] wherein the safety device is configured to relieve an internal pressure within the housing.

[1278] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1279] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1280] an optical sensing unit located on a side of the housing, [1281] wherein the optical sensing unit is configured to detect a reflectance from an environment external to the housing;

[1282] an electronic component located within the housing;

[1283] a gas generating cell located within the housing and adjacent to the electronic component, [1284] wherein the electronic component is configured to activate the gas generating cell to generate gas in response to identifying a location of the ingestible device based on the reflectance;

[1285] a reservoir located within the housing, [1286] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1287] a membrane in contact with the gas generating cell and configured to move or deform into the reservoir by a pressure generated by the gas generating cell; and

[1288] a dispensing outlet placed at the first end of the housing, [1289] wherein the dispensing outlet is configured to deliver the dispensable substance out of the housing from the reservoir.

[1290] In one embodiment, drug delivery is triggered when it encounters the site of release in the GI tract.

[1291] In one embodiment, the one or more environmental sensors measure pH, temperature, transit times, or combinations thereof.

[1292] In some embodiments, releasing the immune modulator (e.g., any of the immune modulators described herein) is dependent on the pH at or in the vicinity of the location. In some embodiments the pH in the jejunum is from 6.1 to 7.2, such as 6.6. In some embodiments the pH in the mid small bowel is from 7.0 to 7.8, such as 7.4. In some embodiments the pH in the ileum is from 7.0 to 8.0, such as 7.5. In some embodiments the pH in the right colon is from 5.7 to 7.0, such as 6.4. In some embodiments the pH in the mid colon is from 5.7 to 7.4, such as 6.6. In some embodiments the pH in the left colon is from 6.3 to 7.7, such as 7.0. In some embodiments, the gastric pH in fasting subjects is from about 1.1 to 2.1, such as from 1.4 to 2.1, such as from 1.1 to 1.6, such as from 1.4 to 1.6. In some embodiments, the gastric pH in fed subjects is from 3.9 to 7.0, such as from 3.9 to 6.7, such as from 3.9 to 6.4, such as from 3.9 to 5.8, such as from 3.9 to 5.5, such as from 3.9 to 5.4, such as from 4.3 to 7.0, such as from 4.3 to 6.7, such as from 4.3 to 6.4, such as from 4.3 to 5.8, such as from 4.3 to 5.5, such as from 4.3 to 5.4. In some embodiments, the pH in the duodenum is from 5.8 to 6.8, such as from 6.0 to 6.8, such as from 6.1 to 6.8, such as from 6.2 to 6.8, such as from 5.8 to 6.7, such as from 6.0 to 6.7, such as from 6.1 to 6.7, such as from 6.2 to 6.7, such as from 5.8 to 6.6, such as from 6.0 to 6.6, such as from 6.1 to 6.6, such as from 6.2 to 6.6, such as from 5.8 to 6.5, such as from 6.0 to 6.5, such as from 6.1 to 6.5, such as from 6.2 to 6.5.

[1293] In some embodiments, releasing the immune modulator is not dependent on the pH at or in the vicinity of the location. In some embodiments, releasing the immune modulator is triggered by degradation of a release component located in the capsule. In some embodiments, the immune modulator is not triggered by degradation of a release component located in the capsule. In some embodiments, wherein releasing the immune modulator is not dependent on enzymatic activity at or in the vicinity of the location. In some embodiments, releasing the immune modulator is not dependent on bacterial activity at or in the vicinity of the location.

[1294] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1295] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1296] a reservoir located within the housing and containing the immune modulator,

[1297] wherein a first end of the reservoir is attached to the first end of the housing;

[1298] a mechanism for releasing the immune modulator from the reservoir;

[1299] and;

[1300] an exit valve configured to allow the immune modulator to be released out of the housing from the reservoir.

[1301] In some embodiments, the ingestible device further comprises:

[1302] an electronic component located within the housing; and

[1303] a gas generating cell located within the housing and adjacent to the electronic component,

[1304] wherein the electronic component is configured to activate the gas generating cell to generate gas.

[1305] In some embodiments, the ingestible device further comprises: a safety device placed within or attached to the housing, [1306] wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.
In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1307] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1308] an electronic component located within the housing;

[1309] a gas generating cell located within the housing and adjacent to the electronic component, [1310] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1311] a reservoir located within the housing, [1312] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1313] an exit valve located at the first end of the housing, [1314] wherein the exit valve is configured to allow the dispensable substance to be released out of the first end of the housing from the reservoir; and

[1315] a safety device placed within or attached to the housing, [1316] wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

[1317] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1318] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1319] an electronic component located within the housing,

[1320] a gas generating cell located within the housing and adjacent to the electronic component, [1321] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1322] a reservoir located within the housing, [1323] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1324] an injection device located at the first end of the housing, [1325] wherein the jet injection device is configured to inject the dispensable substance out of the housing from the reservoir; and

[1326] a safety device placed within or attached to the housing, [1327] wherein the safety device is configured to relieve an internal pressure within the housing.

[1328] In some embodiments, the pharmaceutical composition is an ingestible device, comprising:

[1329] a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1330] an optical sensing unit located on a side of the housing, [1331] wherein the optical sensing unit is configured to detect a reflectance from an environment external to the housing;

[1332] an electronic component located within the housing;

[1333] a gas generating cell located within the housing and adjacent to the electronic component, [1334] wherein the electronic component is configured to activate the gas generating cell to generate gas in response to identifying a location of the ingestible device based on the reflectance;

[1335] a reservoir located within the housing, [1336] wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;

[1337] a membrane in contact with the gas generating cell and configured to move or deform into the reservoir by a pressure generated by the gas generating cell; and

[1338] a dispensing outlet placed at the first end of the housing, [1339] wherein the dispensing outlet is configured to deliver the dispensable substance out of the housing from the reservoir.

[1340] In some embodiments, the pharmaceutical composition is an ingestible device as disclosed in U.S. Patent Application Ser. No. 62/385,553, incorporated by reference herein in its entirety.

[1341] In some embodiments, the pharmaceutical composition is an ingestible device as disclosed in the following applications, each of which is incorporated by reference herein in its entirety:

U.S. Ser. No. 14/460,893; 15/514,413; 62/376,688; 62/385,344; 62/478,955; 62/434,188; 62/434,320; 62/431,297; 62/434,797; 62/480,187; 62/502,383; and 62/540,873.

[1342] In some embodiments, the pharmaceutical composition is an ingestible device comprising a localization mechanism as disclosed in international patent application PCT/US2015/052500, incorporated by reference herein in its entirety.

[1343] In some embodiments, the pharmaceutical composition is not a dart-like dosage form.

[1344] In some embodiments provided herein is an ingestible device, comprising:

[1345] an immune modulator;

[1346] one or more processing devices; and

[1347] one more machine readable hardware storage devices storing instructions that are executable by the one or more processing devices to determine a location of the ingestible device in a portion of a GI tract of a subject to an accuracy of at least 85%. In some embodiments, the accuracy is at least 90%. In some embodiments, the accuracy is at least 95%. In some embodiments, the accuracy is at least 97%. In some embodiments, the accuracy is at least 98%. In some embodiments, the accuracy is at least 99%. In some embodiments, the accuracy is 100%. In some embodiments, the portion of the GI tract of the subject comprises the duodenum. In some embodiments, the portion of the GI tract of the subject comprises the jejunum. In some embodiments, the portion of the GI tract of the subject comprises the terminal ileum, cecum and colon. In some embodiments, the ingestible device further comprises first and second light sources, wherein the first light source is configured to emit light at a first wavelength, and the second light source is configured to emit light at a second wavelength different from the first wavelength. In some embodiments, the ingestible device further comprises first and second detectors, wherein the first detector is configured to detect light at the first wavelength, and the second detector is configured to detect light at the second wavelength.

[1348] In some embodiments, provided herein is an ingestible device, comprising:

[1349] an immune modulator;

[1350] one or more processing devices; and

[1351] one more machine readable hardware storage devices storing instructions that are executable by the one or more processing devices to determine that the ingestible device is in the cecum of a subject to an accuracy of at least 70%. In some embodiments, the accuracy is at least 75%. In some embodiments, the accuracy is at least 80%. In some embodiments, the accuracy is at least 85%. In some embodiments, the accuracy is at least 88%. In some embodiments, the accuracy is at least 89%.

[1352] In some embodiments, provided herein is an ingestible device, comprising:

[1353] an immune modulator;

[1354] one or more processing devices; and

[1355] one more machine readable hardware storage devices storing instructions that are executable by the one or more processing devices to transmit data to a device capable of implementing the data to determine a location of the medical device in a portion of a GI tract of a subject to an accuracy of at least 85%. In some embodiments, the accuracy is at least 90%. In some embodiments, the accuracy is at least 95%. In some embodiments, the accuracy is at least 97%. In some embodiments, the accuracy is at least 98%. In some embodiments, the accuracy is at least 99%. In some embodiments, the accuracy is 100%. In some embodiments, the portion of the GI tract of the subject comprises the duodenum. In some embodiments, the portion of the GI tract of the subject comprises the jejunum. In some embodiments, the portion of the GI tract of the subject comprises the terminal ileum, cecum and colon. In some embodiments, the ingestible device further comprises first and second light sources, wherein the first light source is configured to emit light at a first wavelength, and the second light source is configured to emit light at a second wavelength different from the first wavelength. In some embodiments, the ingestible device further comprises first and second detectors, wherein the first detector is configured to detect light at the first wavelength, and the second detector is configured to detect light at the second wavelength. In some embodiments, the data comprise intensity data for at least two different wavelengths of light.

[1356] In some embodiments, provided herein is an ingestible device, comprising:

[1357] an immune modulator;

[1358] one or more processing devices; and

[1359] one more machine readable hardware storage devices storing instructions that are executable by the one or more processing devices to transmit data to an external device capable of implementing the data to determine that the ingestible device is in the cecum of subject to an accuracy of at least 70%. In some embodiments, the accuracy is at least 75%. In some embodiments, the accuracy is at least 80%. In some embodiments, the accuracy is at least 85%. In some embodiments, the accuracy is at least 88%. In some embodiments, the accuracy is at least 89%.

[1360] In some embodiments, provided herein is a method of treating an inflammatory disease or condition arising in a tissue that originates from the endoderm in a subject, comprising: releasing an immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release, wherein the method comprises administering orally to the subject an ingestible device as disclosed herein, the method further comprising determining a location of the ingestible medical device in a portion of a GI tract of the subject to an accuracy of at least 85%. In some embodiments, the accuracy is at least 90%. In some embodiments, the accuracy is at least 95%. In some embodiments, the accuracy is at least 97%. In some embodiments, the accuracy is at least 98%. In some embodiments, the accuracy is at least 99%. In some embodiments, the accuracy is 100%. In some embodiments, the portion of the GI tract of the subject comprises the duodenum. In some embodiments, the portion of the GI tract of the subject comprises the jejunum. In some embodiments, the portion of the GI tract of the subject comprises the terminal ileum, cecum and colon. In some embodiments, determining the location of the ingestible device within the GI tract of a subject comprises determining reflected light signals within the GI tract, wherein the reflected signals comprise light of at least two different wavelengths. In some embodiments, the reflected signals comprise light of at least three different wavelengths. In some embodiments, the reflected light comprise first and second wavelengths; the first wavelength is between 495-600 nm; and the second wavelength is between 400-495 nm. In some embodiments, the first and second wavelengths are separated by at least 50 nm.

[1361] In some embodiments, provided herein is a method of treating an inflammatory disease or condition arising in a tissue originating from the endoderm in a subject, comprising: releasing an immune modulator at a location in the gastrointestinal tract of the subject that is proximate to the intended site of release, wherein the method comprises administering orally to the subject an ingestible device as disclosed herein, the method further comprising determining a location of the ingestible medical device within the GI tract of the subject based on measured reflected light signals within the GI tract, where the reflected signals comprise light of at least two different wavelengths. In some embodiments, the reflected signals comprise light of at least three different wavelengths. In some embodiments, the at least two different wavelengths comprise first and second wavelengths; the first wavelength is between 495-600 nm; and the second wavelength is between 400-495 nm. In some embodiments, the first and second wavelengths are separated by at least 50 nm.

[1362] In some embodiments, provided herein is an ingestible device, comprising:

[1363] a housing;

[1364] a gas generating cell located within the housing; and

[1365] a storage reservoir located within the housing,

[1366] wherein the storage reservoir stores an immune modulator, and an opening in the housing is configured to allow the immune modulator to be released out of the housing from the storage reservoir via an opening in the ingestible device.

[1367] In some embodiments, the housing is defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1368] wherein an electronic component is located within the housing and the gas generating cell is adjacent to the electronic component,

[1369] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1370] wherein a first end of the storage reservoir is connected to the first end of the housing;

[1371] wherein an exit valve is located at the first end of the housing and is configured to allow the immune modulator to be released out of the first end of the housing; and wherein the ingestible device further comprises a safety device placed within or attached to the housing,

[1372] wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

[1373] In some embodiments, provided herein is an ingestible device, comprising:

[1374] a gas generating cell located within the housing;

[1375] a storage reservoir located within the housing,

[1376] wherein the storage reservoir stores an immune modulator; and

[1377] an injection device configured to inject the immune modulator out of the housing from the storage reservoir via an opening in the ingestible device.

[1378] In some embodiments, the housing is defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1379] wherein an electronic component is located within the housing and the gas generating cell is adjacent to the electronic component,

[1380] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1381] wherein a first end of the storage reservoir is connected to the first end of the housing; wherein the injection device is located at the first end of the housing and is configured to inject the immune modulator out of the housing via an opening in the ingestible device; and and wherein the ingestible device further comprises a safety device placed within or attached to the housing, [1382] wherein the safety device is configured to relieve an internal pressure within the housing.

[1383] In some embodiments, provided herein is an ingestible device, comprising:

[1384] a housing;

[1385] an optical sensing unit supported by a side of the housing,

[1386] wherein the optical sensing unit is configured to detect a reflectance from an environment external to the housing;

[1387] a gas generating cell located within the housing,

[1388] wherein the ingestible device is configured so that, in response to identifying a location of the ingestible device based on a reflectance detected by the optical sensing unit, the gas generating cell generates a gas;

[1389] a storage reservoir located within the housing,

[1390] wherein the storage reservoir stores an immune modulator;

[1391] and wherein the ingestible device is configured so that, when the gas generating cell generates the gas, the immune modulator is delivered out of the housing from the storage reservoir via an opening in the ingestible device.

[1392] In some embodiments, the housing is defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;

[1393] wherein the optical sensing unit is supported by the side of the housing,

[1394] wherein the ingestible device further comprises an electronic component located within the housing;

[1395] wherein the gas generating cell is adjacent to the electronic component,

[1396] wherein the electronic component is configured to activate the gas generating cell to generate gas;

[1397] wherein a first end of the storage reservoir is connected to the first end of the housing;

[1398] wherein the ingestible device further comprises a membrane in contact with the gas generating cell and configured to move or deform into the storage reservoir by a pressure generated by the gas generating cell; and

[1399] wherein the ingestible device further comprises a dispensing outlet placed at the first end of the housing and configured to deliver the immune modulator out of the housing.

[1400] In some embodiments of any ingestible device disclosed herein comprising an immune modulator, the immune modulator is present in a therapeutically effective amount.

[1401] In case of conflict between the present specification and any subject matter incorporated by reference herein, the present specification, including definitions, will control.

Devices and Methods for Detection of Analytes in GI Tract

[1402] Detection of certain analytes in the GI tract may be useful in the identification of the nature and severity of the disease, in accurately locating the site(s) of disease, and in assessing patient response to a therapeutic agent. The appropriate therapeutic agent may accordingly be released at the correct locations(s), dosage, or timing for the disease. As discussed further herein, analytes may include biomarkers associated with a disease or associated with patient response and/or therapeutic agents previously administered to treat the disease. In some embodiments, the disclosure provides an ingestible device for detecting an analyte in a sample, the ingestible device comprising a sampling chamber that is configured to hold a composition comprising: (1) a plurality of donor particles, each of the plurality of donor particles comprising a photosensitizer and having coupled thereto a first antigen-binding agent that binds to the analyte, wherein the photosensitizer, in its excited state, is capable of generating singlet oxygen; and (2) a plurality of acceptor particles, each of the plurality of acceptor particles comprising a chemiluminescent compound and having coupled thereto a second antigen-binding agent that binds to the analyte, wherein the chemiluminescent compound is capable of reacting with singlet oxygen to emit luminescence. In some embodiments, the first and the second analyte-binding agents are antigen-binding agents (e.g., antibodies). In some embodiments, the first and the second antigen-binding agents bind to the same epitope of the analyte (e.g., a protein). In some embodiments, the first and the second antigen-binding agents bind to separate epitopes of the analyte (e.g., a protein) that spatially overlap. In some embodiments, the first and the second antigen-binding agents bind to the separate epitopes of the analyte (e.g., a protein) that do not spatially overlap.

[1403] In some embodiments, this disclosure provides an ingestible device for detecting an analyte in a sample, the ingestible device comprising a sampling chamber that is configured to hold an absorbable material (e.g., an absorbable pad or sponge) having absorbed therein a composition comprising: (1) a plurality of donor particles, each of the plurality of donor particles comprising a photosensitizer and having coupled thereto a first antigen-binding agent that binds to the analyte, wherein the photosensitizer, in its excited state, is capable of generating singlet oxygen; and (2) a plurality of acceptor particles, each of the plurality of acceptor particles comprising a chemiluminescent compound and having coupled thereto a second antigen-binding agent that binds to the analyte, wherein the chemiluminescent compound is capable of reacting with singlet oxygen to emit luminescence. In some embodiments, the first and the second analyte-binding agents are antigen-binding agents (e.g., antibodies). In some embodiments, the first and the second antigen-binding agents bind to the same epitope of the analyte (e.g., a protein). In some embodiments, the first and the second antigen-binding agents bind to separate epitopes of the analyte (e.g., a protein) that spatially overlap. In some embodiments, the first and the second antigen-binding agents bind to the separate epitopes of the analyte (e.g., a protein) that do not spatially overlap.

[1404] In certain embodiments, the disclosure provides a kit comprising an ingestible device as described herein. In some embodiments, the kit further comprises instructions, e.g., for detecting or quantifying an analyte in a sample.

[1405] In some embodiments, the disclosure provides methods for determining an analyte in a sample. In certain embodiments, this disclosure provides a method of detecting an analyte in a fluid sample of a subject, comprising: (1) providing an ingestible device; (2) transferring the fluid sample of the subject into the sampling chamber of the ingestible device in vivo; (3) irradiating the composition held in the sampling chamber of the ingestible device with light to excite the photosensitizer; and (4) measuring total luminescence or rate of change of luminescence emitted from the composition held in the sampling chamber of the ingestible device as a function of time, thereby determining the level of the analyte in the fluid sample. In some embodiments, the method further comprises comparing the level of the analyte in the fluid sample with the level of analyte in a reference sample (e.g., a reference sample obtained from a healthy subject). In some embodiments, the level of the analyte in the sample is used to diagnose and/or monitor a disease or disorder in the subject.

[1406] In some embodiments, the disclosure provides a method of detecting an analyte in a fluid sample of a subject, comprising: (1) providing an ingestible device, the device comprising a sampling chamber that is configured to hold an absorbable material (e.g., an absorbable pad or sponge) having absorbed therein a composition, as described herein; (2) transferring the fluid sample of the subject into the sampling chamber of the ingestible device in vivo; (3) fully or partially saturating the absorbable material held in the sampling chamber of the ingestible device with the fluid sample; (4) irradiating the absorbable material held in the sampling chamber of the ingestible device with light to excite the photosensitizer; and (5) measuring total luminescence or rate of change of luminescence emitted from the composition held in the sampling chamber of the ingestible device as a function of time, thereby determining the level of the analyte in the fluid sample. In some embodiments, the method further comprises comparing the level of the analyte in the fluid sample with the level of analyte in a reference sample (e.g., a reference sample obtained from a healthy subject). In some embodiments, the level of the analyte in the sample is used to diagnose and/or monitor a disease or disorder in the subject.

[1407] In some embodiments, the disclosure provides a method of assessing or monitoring the need to treat a subject suffering from or at risk of overgrowth of bacterial cells in the gastrointestinal (GI) tract, comprising: (1) providing an ingestible device for detecting an analyte; (2) transferring a fluid sample from the GI tract of the subject into the sampling chamber of the ingestible device in vivo; (3) irradiating the composition held in the sampling chamber of the ingestible device with light to excite the photosensitizer; (4) measuring total luminescence or rate of change of luminescence emitted from the composition held in the sampling chamber of the ingestible device as a function of time; (5) correlating the total luminescence or the rate of change of luminescence as a function of time measured in step (4) to the amount of the analyte in the fluid sample; and (6) correlating the amount of the analyte in the fluid sample to the number of viable bacterial cells in the fluid sample. In some embodiments, a number of viable bacterial cells determined in step (6) greater than a control number of viable bacterial cells, indicates a need for treatment (e.g., with an antibiotic agent described herein). In some embodiments, the control number of viable bacterial cells is 10.sup.3, 10.sup.4, 10.sup.5, 10.sup.6, 10.sup.7, 10.sup.8, 10.sup.9, or more. For example, in some embodiments, a number of viable bacterial cells determined in step (6) greater that about 10.sup.3 CFU/mL indicates a need for treatment. In some embodiments, a number of viable bacterial cells determined in step (6) greater that about 10.sup.4 CFU/mL indicates a need for treatment. In some embodiments, a number of the viable bacterial cells determined in step (6) greater than about 105 CFU/mL indicates a need for treatment, e.g., with an antibiotic agent as described herein. In some embodiments, a number of viable bacterial cells determined in step (6) greater that about 10.sup.6 or more CFU/mL indicates a need for treatment.

[1408] In some embodiments, the total luminescence or the rate of change of luminescence as a function of time of the sponge is measured over multiple time points for an extended period of time in step (4). For instance, in some embodiments, the total luminescence or rate of change of luminescence as a function of time of the sample is measured continuously for a period of 0-1800 minutes, 0-1600 minutes, 0-1500 minutes, 0-1440 minutes, 0-1320 minutes, 0-1000 minutes, 0-900 minutes, 0-800 minutes, 0-700 minutes, 0-600 minutes, 0-500 minutes, 0-400 minutes, 0-350 minutes, 0-330 minutes, 0-300 minutes, 0-270 minutes, or 0-220 minutes. In some embodiments, the total luminescence or the rate of change of luminescence as a function of time of said sample is measured continuously for a period of 0-330 minutes. In some embodiments, the method is performed in vivo. In some embodiments, the method includes communicating the results of the onboard assay(s) to an ex vivo receiver. In some embodiments, the total luminescence or the rate of change of luminescence as a function of time of the sponge is measured over multiple time points for an extended period of time in step (5). For instance, in some embodiments, the total luminescence or rate of change of luminescence as a function of time of the sample is measured continuously for a period of 0-1800 minutes, 0-1600 minutes, 0-1500 minutes, 0-1440 minutes, 0-1320 minutes, 0-1000 minutes, 0-900 minutes, 0-800 minutes, 0-700 minutes, 0-600 minutes, 0-500 minutes, 0-400 minutes, 0-350 minutes, 0-330 minutes, 0-300 minutes, 0-270 minutes, or 0-220 minutes. In some embodiments, the total luminescence or the rate of change of luminescence as a function of time of said sample is measured continuously for a period of 0-330 minutes. In some embodiments, the method is performed in vivo. In some embodiments, the method includes communicating the results of the onboard assay(s) to an ex vivo receiver.

[1409] In some embodiments, the disclosure provides a method of assessing or monitoring the need to treat a subject suffering from or at risk of overgrowth of bacterial cells in the gastrointestinal tract, comprising: (1) providing an ingestible device for detecting an analyte, the device comprising a sampling chamber that is configured to hold an absorbable material (e.g., an absorbable pad or sponge) having absorbed therein a composition, as described herein; (2) transferring a fluid sample from the GI tract of the subject into the sampling chamber of the ingestible device in vivo; (3) fully or partially saturating the absorbable material held in the sampling chamber of the ingestible device with the fluid sample; (4) irradiating the absorbable material held in the sampling chamber of the ingestible device with light to excite the photosensitizer; (5) measuring total luminescence or rate of change of luminescence emitted from the composition held in the sampling chamber of the ingestible device as a function of time; (6) correlating the total luminescence or the rate of change of luminescence as a function of time measured in step (5) to the amount of the analyte in the fluid sample; and (7) correlating the amount of the analyte in the fluid sample to the number of viable bacterial cells in the fluid sample. In some embodiments, a number of viable bacterial cells determined in step (7) greater than a control number of viable bacterial cells indicates a need for treatment (e.g., with an antibiotic agent described herein). In some embodiments, the control number of viable bacterial cells is 10.sup.3, 10.sup.4, 10.sup.5, 10.sup.6, 10.sup.7, 10.sup.8, 10.sup.9, or more. For example, in some embodiments, a number of viable bacterial cells determined in step (7) greater that about 10.sup.3 CFU/mL indicates a need for treatment. In some embodiments, a number of viable bacterial cells determined in step (7) greater that about 10.sup.4 CFU/mL indicates a need for treatment. In some embodiments, a number of the viable bacterial cells determined in step (7) greater than about 10.sup.5 CFU/mL indicates a need for treatment, e.g., with an antibiotic agent as described herein. In some embodiments, a number of viable bacterial cells determined in step (7) greater that about 10.sup.6 or more CFU/mL indicates a need for treatment.

[1410] In some embodiments, the disclosure, provides a method of measuring the presence, absence or amount of one or more analytes from one or more samples in the gastrointestinal tract. In some embodiments the one or more analytes are measured multiple times, for example, at different time points or at different locations. In one embodiment, a single device measures one or more analytes or more time points or locations; thereby creating a molecular map of a physiological region. Measurements can be taken at any location in the gastrointestinal tract. For example, in one aspect, analytes from samples from one or more of the duodenum, jejunum, ileum, ascending colon, transverse colon or descending colon can be measured to create a molecular map of the small and large intestine. In one aspect, the sample is from the duodenum. In one aspect, In one aspect, the sample is from the jejunum. In one aspect, the sample is from the ileum. In one aspect, the sample is from the ascending colon. In one aspect, the sample is from the transverse colon. In one aspect, the sample is from the descending colon.

[1411] In another aspect, a series of measurements can be taken over a shorter distance of the gastrointestinal tract (e.g., the ileum) to create a higher resolution molecular map. In some embodiments, previous endoscopic imaging may identify a diseased area for molecular mapping. For example, a gastroenterologist may use imaging (e.g., an endoscope equipped with a camera) to identify the presence of Crohn's Disease in the ileum and cecum of a patient, and the methods and techniques herein may be used to measure inflammation-associated analytes in this diseased area of the patient. In a related embodiment, the inflammation-associated analytes, or any analyte, may be measured every one or more days to monitor disease flare-ups, or response to therapeutics.

Analytes

[1412] The compositions and methods described herein can be used to detect, analyze, and/or quantitate a variety of analytes in a human subject. Analyte as used herein refers to a compound or composition to be detected in a sample. Exemplary analytes suitable for use herein include those described in U.S. Pat. No. 6,251,581, which is incorporated by reference herein in its entirety. Broadly speaking, an analyte can be any substance (e.g., a substance with one or more antigens) capable of being detected. An exemplary and non-limiting list of analytes includes ligands, proteins, blood clotting factors, hormones, cytokines, polysaccharides, mucopolysaccharides, microorganisms (e.g., bacteria), microbial antigens, and therapeutic agents (including fragments and metabolites thereof).

[1413] For instance, the analyte may be a ligand, which is monovalent (monoepitopic) or polyvalent (polyepitopic), usually antigenic or haptenic, and is a single compound or plurality of compounds which share at least one common epitopic or determinant site. The analyte can be a part of a cell such as bacteria or a cell bearing a blood group antigen such as A, B, D, etc., a human leukocyte antigen (HLA), or other cell surface antigen, or a microorganism, e.g., bacterium (e.g. a pathogenic bacterium), a fungus, protozoan, or a virus (e.g., a protein, a nucleic acid, a lipid, or a hormone). In some embodiments, the analyte can be a part of an exosome (e.g., a bacterial exosome). In some embodiments, the analyte is derived from a subject (e.g., a human subject). In some embodiments, the analyte is derived from a microorganism present in the subject. In some embodiments, the analyte is a nucleic acid (e.g., a DNA molecule or a RNA molecule), a protein (e.g., a soluble protein, a cell surface protein), or a fragment thereof, that can be detected using any of the devices and methods provided herein.

[1414] The polyvalent ligand analytes will normally be poly(amino acids), i.e., a polypeptide (i.e., protein) or a peptide, polysaccharides, nucleic acids (e.g., DNA or RNA), and combinations thereof. Such combinations include components of bacteria, viruses, chromosomes, genes, mitochondria, nuclei, cell membranes, and the like.

[1415] In some embodiments, the polyepitopic ligand analytes have a molecular weight of at least about 5,000 Da, more usually at least about 10,000 Da. In the poly(amino acid) category, the poly(amino acids) of interest may generally have a molecular weight from about 5,000 Da to about 5,000,000 Da, more usually from about 20,000 Da to 1,000,000 Da; among the hormones of interest, the molecular weights will usually range from about 5,000 Da to 60,000 Da.

[1416] In some embodiments, the monoepitopic ligand analytes generally have a molecular weight of from about 100 to 2,000 Da, more usually from 125 to 1,000 Da.

[1417] A wide variety of proteins may be considered as to the family of proteins having similar structural features, proteins having particular biological functions, proteins related to specific microorganisms, particularly disease causing microorganisms, etc. Such proteins include, for example, immunoglobulins, cytokines, enzymes, hormones, cancer antigens, nutritional markers, tissue specific antigens, etc.

[1418] In some embodiments, the analyte is a protein. In some embodiments, the analyte is a protein, e.g., an enzyme (e.g., a hemolysin, a protease, a phospholipase), a soluble protein, an exotoxin. In some embodiments, the analyte is a fragment of a protein, a peptide, or an antigen. In some embodiments, the analyte is a peptide of at least 5 amino acids (e.g., at least 6, at least 7, at least 8, at least 9, at least 10, at least 25, at least, 50, or at least 100 amino acids). Exemplary lengths include 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 50, 75, or 100 amino acids. Exemplary classes of protein analytes include, but are not limited to: protamines, histones, albumins, globulins, scleroproteins, phosphoproteins, mucoproteins, chromoproteins, lipoproteins, nucleoproteins, glycoproteins, T-cell receptors, proteoglycans, cell surface receptors, membrane-anchored proteins, transmembrane proteins, secreted proteins, HLA, and unclassified proteins.

[1419] In some embodiments, the analyte is an affimer (see, e.g., Tiede et al. (2017) eLife 6: e24903, which is expressly incorporated herein by reference).

[1420] Exemplary analytes include: Prealbumin, Albumin, .sub.1-Lipoprotein, .sub.1-Antitrypsin, .sub.1-Glycoprotein, Transcortin, 4.6S-Postalbumin, .sub.1-glycoprotein, .sub.1X-Glycoprotein, Thyroxin-binding globulin, Inter--trypsin-inhibitor, Gc-globulin (Gc 1-1, Gc 2-1, Gc 2-2), Haptoglobin (Hp 1-1, Hp 2-1, Hp 2-2), Ceruloplasmin, Cholinesterase, .sub.2-Lipoprotein(s), Myoglobin, C-Reactive Protein, .sub.2-Macroglobulin, .sub.2-HS-glycoprotein, Zn-.sub.2-glycoprotein, .sub.2-Neuramino-glycoprotein, Erythropoietin, -lipoprotein, Transferrin, Hemopexin, Fibrinogen, Plasminogen, .sub.2-glycoprotein I, .sub.2-glycoprotein II, Immunoglobulin G (IgG) or G-globulin, Immunoglobulin A (IgA) or A-globulin, Immunoglobulin M (IgM) or M-globulin, Immunoglobulin D (IgD) or D-Globulin (D), Immunoglobulin E (IgE) or E-Globulin (E), Free K and X light chains, and Complement factors: C1, (C1q, C1r, C1s, C2, C3 (.sub.1A, .sub.2D), C4, C5, C6, C7, C8, C9.

[1421] Additional examples of analytes include tumor necrosis factor- (TNF), interleukin-12 (IL-12), IL-23, IL-6, 21 integrin, 41 integrin, 47 integrin, integrin 41 (VLA-4), E-selectin, ICAM-1, 51 integrin, 41 integrin, VLA-4, 21 integrin, 53 integrin, 55 integrin, IIb3 integrin, MAdCAM-1, SMAD7, JAK1, JAK2, JAK3, TYK-2, CHST15, IL-1, IL-1, IL-1, IL-18, IL-36, IL-36, IL-36, IL-38, IL-33, IL-13, CD40L, CD40, CD3, CD3, CD3, CD3, TCR, TCR, TCR, TCR, TCR, CD14, CD20, CD25, IL-2, IL-2 chain, IL-2 chain, CD28, CD80, CD86, CD49, MMP1, CD89, IgA, CXCL10, CCL11, an ELR chemokine, CCR2, CCR9, CXCR3, CCR3, CCR5, CCL2, CCL8, CCL16, CCL25, CXCR1m CXCR2m CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, and CXCL8, and a nucleic acid (e.g., mRNA) encoding any of the same.

[1422] In some embodiments, the analyte is a blood clotting factor. Exemplary blood clotting factors include, but are not limited to:

TABLE-US-00026 International designation Name I Fibrinogen II Prothrombin IIa Thrombin III Tissue thromboplastin V and VI Proaccelerin, accelerator globulin VII Proconvertin VIII Antihemophilic globulin (AHG) IX Christmas factor plasma thromboplastin component (PTC) X Stuart-Prower factor, autoprothrombin III XI Plasma thromboplastin antecedent (PTA) XII Hagemann factor XIII Fibrin-stabilizing factor

[1423] In some embodiments, the analyte is a hormone. Exemplary hormones include, but are not limited to: Peptide and Protein Hormones, Parathyroid hormone, (parathromone), Thyrocalcitonin, Insulin, Glucagon, Relaxin, Erythropoietin, Melanotropin (melancyte-stimulating hormone; intermedin), Somatotropin (growth hormone), Corticotropin (adrenocorticotropic hormone), Thyrotropin, Follicle-stimulating hormone, Luteinizing hormone (interstitial cell-stimulating hormone), Luteomammotropic hormone (luteotropin, prolactin), Gonadotropin (chorionic gonadotropin), Secretin, Gastrin, Angiotensin I and II, Bradykinin, and Human placental lactogen, thyroxine, cortisol, triiodothyronine, testosterone, estradiol, estrone, progestrone, luteinizing hormone-releasing hormone (LHRH), and immunosuppressants such as cyclosporin, FK506, mycophenolic acid, and so forth.

[1424] In some embodiments, the analyte is a peptide hormone (e.g., a peptide hormone from the neurohypophysis). Exemplary peptide hormones from the neurohypophysis include, but are not limited to: Oxytocin, Vasopressin, and releasing factors (RF) (e.g., corticotropin releasing factor (CRF), luteinizing hormone releasing factor (LRF), thyrotropin releasing factor (TRY), Somatotropin-RF, growth hormone releasing factor (GRF), follicle stimulating hormone-releasing factor (FSH-RF), prolactin inhibiting factor (PIF), and melanocyte stimulating hormone inhibiting factor (MIF)).

[1425] In some embodiments, the analyte is a cytokine or a chemokine. Exemplary cytokines include, but are not limited to: interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), epidermal growth factor (EGF), tumor necrosis factor (TNF, e.g., TNF- or TNF-), and nerve growth factor (NGF).

[1426] In some embodiments, the analyte is a cancer antigen. Exemplary cancer antigens include, but are not limited to: prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), -fetoprotein, Acid phosphatase, CA19.9, and CA125.

[1427] In some embodiments, the analyte is a tissue-specific antigen. Exemplary tissue specific antigens include, but are not limited to: alkaline phosphatase, myoglobin, CPK-MB, calcitonin, and myelin basic protein.

[1428] In some embodiments, the analyte is a mucopolysaccharide or a polysaccharide.

[1429] In some embodiments, the analyte is a microorganism, or a molecule derived from or produced by a microorganism (e.g., a bacteria, a virus, prion, or a protozoan). For example, in some embodiments, the analyte is a molecule (e.g., an protein or a nucleic acid) that is specific for a particular microbial genus, species, or strain (e.g., a specific bacterial genus, species, or strain). In some embodiments, the microorganism is pathogenic (i.e., causes disease). In some embodiments, the microorganism is non-pathogenic (e.g., a commensal microorganism). Exemplary microorganisms include, but are not limited to:

TABLE-US-00027 Corynebacteria Corynebacterium diphtheria Pneumococci Diplococcus pneumoniae Streptococci Streptococcus pyrogenes Streptococcus salivarus Staphylococci Staphylococcus aureus Staphylococcus albus Neisseria Neisseria meningitidis Neisseria gonorrhea Enterobacteriaciae Escherichia coli Aerobacter aerogenes The coliform Klebsiella pneumoniae bacteria Salmonella typhosa Salmonella choleraesuis The Salmonellae Salmonella typhimurium Shigella dysenteria Shigella schmitzii Shigella arabinotarda The Shigellae Shigella flexneri Shigella boydii Shigella sonnei Other enteric bacilli Proteus vulgaris Proteus mirabilis Proteus species Proteus morgani Pseudomonas aeruginosa Alcaligenes faecalis Vibrio cholerae Hemophilus-Bordetella group Rhizopus oryzae Hemophilus influenza, H. ducryi Rhizopus arrhizua Phycomycetes Hemophilus hemophilus Rhizopus nigricans Hemophilus aegypticus Sporotrichum schenkii Hemophilus parainfluenza Flonsecaea pedrosoi Bordetella pertussis Fonsecacea compact Pasteurellae Fonsecacea dermatidis Pasteurella pestis Cladosporium carrionii Pasteurella tulareusis Phialophora verrucosa Brucellae Aspergillus nidulans Brucella melltensis Madurella mycetomi Brucella abortus Madurella grisea Brucella suis Allescheria boydii Aerobic Spore-forming Bacilli Phialophora jeanselmei Bacillus anthracis Microsporum gypseum Bacillus subtilis Trichophyton mentagrophytes Bacillus megaterium Keratinomyces ajelloi Bacillus cereus Microsporum canis Anaerobic Spore-forming Bacilli Trichophyton rubrum Clostridium botulinum Microsporum adouini Clostridium tetani Viruses Clostridium perfringens Adenoviruses Clostridium novyi Herpes Viruses Clostridium septicum Herpes simplex Clostridium histoyticum Varicella (Chicken pox) Clostridium tertium Herpes Zoster (Shingles) Clostridium bifermentans Virus B Clostridium sporogenes Cytomegalovirus Mycobacteria Pox Viruses Mycobacterium tuberculosis hominis Variola (smallpox) Mycobacterium bovis Vaccinia Mycobacterium avium Poxvirus bovis Mycobacterium leprae Paravaccinia Mycobacterium paratuberculosis Molluscum contagiosum Actinomycetes (fungus-ike bacteria) Picornaviruses Actinomyces Isaeli Poliovirus Actinomyces bovis Coxsackievirus Actinomyces naeslundii Echoviruses Nocardia asteroides Rhinoviruses Nocardia brasiliensis Myxoviruses The Spirochetes Influenza(A, B, and C) Treponema pallidum Parainfluenza (1-4) Treponema pertenue Mumps Virus Spirillum minus Streptobacillus monoiliformis Newcastle Disease Virus Treponema carateum Measles Virus Borrelia recurrentis Rinderpest Virus Leptospira icterohemorrhagiae Canine Distemper Virus Leptospira canicola Respiratory Syncytial Virus Trypanasomes Rubella Virus Mycoplasmas Arboviruses Mycoplasma pneumoniae Other pathogens Eastern Equine Encephalitis Virus Listeria monocytogenes Western Equine Encephalitis Virus Erysipeothrix rhusiopathiae Sindbis Virus Streptobacillus moniliformis Chikugunya Virus Donvania granulomatis Semliki Forest Virus Entamoeba histolytica Mayora Virus Plasmodium falciparum St. Louis Encephalitis Plasmodium japonicum California Encephalitis Virus Bartonella bacilliformis Colorado Tick Fever Virus Rickettsia (bacteria-like parasites) Yellow Fever Virus Rickettsia prowazekii Dengue Virus Rickettsia mooseri Reoviruses Rickettsia rickettsii Reovirus Types 1-3 Rickettsia conori Retroviruses Rickettsia australis Human Immunodeficiency Rickettsia sibiricus Viruses I and II (HTLV) Rickettsia akari Human T-cell Lymphotrophic Rickettsia tsutsugamushi Virus I & II (HIV) Rickettsia burnetti Hepatitis Rickettsia quintana Hepatitis A Virus Chlamydia (unclassifiable parasites Hepatitis B Virus bacterial/viral) Hepatitis C Virus Chlamydia agents (naming uncertain) Tumor Viruses Chlamydia trachomatis Fungi Rauscher Leukemia Virus Cryptococcus neoformans Gross Virus Blastomyces dermatidis Maloney Leukemia Virus Histoplasma capsulatum Coccidioides immitis Human Papilloma Virus Paracoccidioides brasliensis Candida albicans Aspergillus fumigatus Mucor corymbifer (Absidia corymbifera)

[1430] In some embodiments, the analyte is a bacterium. Exemplary bacteria include, but are not limited to: Escherichia coli (or E. coli), Bacillus anthracis, Bacillus cereus, Clostridium botulinum, Clostridium difficile, Yersinia pestis, Yersinia enterocolitica, Francisella tularensis, Brucella species, Clostridium perfringens, Burkholderia mallei, Burkholderia pseudomallei, Staphylococcus species, Mycobacterium species, Group A Streptococcus, Group B Streptococcus, Streptococcus pneumoniae, Helicobacter pylori, Salmonella enteritidis, Mycoplasma hominis, Mycoplasma orale, Mycoplasma salivarium, Mycoplasma fermentans, Mycoplasma pneumoniae, Mycobacterium bovis, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium leprae, Rickettsia rickettsii, Rickettsia akari, Rickettsia prowazekii, Rickettsia canada, Bacillus subtilis, Bacillus subtilis niger, Bacillus thuringiensis, Coxiella burnetti, Faecalibacterium prausnitzii (also known as Bacteroides praussnitzii), Roseburia hominis, Eubacterium rectale, Dialister invisus, Ruminococcus albus, Ruminococcus callidus, and Ruminococcus bromii. Additional exemplary bacteria include bacteria of the phyla Firmicutes (e.g., Clostridium clusters XIVa and IV), bacteria of the phyla Bacteroidetes (e.g., Bacteroides fragilis or Bacteroides vulgatus), and bacteria of the phyla Actinobacteria (e.g., Coriobacteriaceae spp. or Bifidobacterium adolescentis). Bacteria of the Clostridium cluster XIVa includes species belonging to, for example, the Clostridium, Ruminococcus, Lachnospira, Roseburia, Eubacterium, Coprococcus, Dorea, and Butyrivibrio genera. Bacteria of the Clostridium cluster IV includes species belonging to, for example, the Clostridium, Ruminococcus, Eubacterium and Anaerofilum genera. In some embodiments, the analyte is Candida, e.g., Candida albicans. In some embodiments, the analyte is a byproduct from a bacterium or other microorganism, e.g., helminth ova, enterotoxin (Clostridium difficile toxin A; TcdA) or cytotoxin (Clostridium difficile toxin B; TcdB).

[1431] In some embodiments, the bacterium is a pathogenic bacterium. Non-limiting examples of pathogenic bacteria belong to the genera Bacillus, Bordetella, Borrelia, Brucella, Campylobacter, Chlamydia, Chlamydophila, Clostridium, Corynebacterium, Enterobacter, Enterococcus, Escherichia, Francisella, Haemophilus, Helicobacter, Legionella, Leptospira, Listeria, Mycobacterium, Mycoplasma, Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella, Staphylococcus, Streptococcus, Treponema, Vibrio, and Yersinia. Non-limiting examples of specific pathogenic bacterial species include a strain of Bacillus anthracis, a strain of a strain of Bordetella pertussis, a strain of a strain of Borrelia burgdorferi, a strain of a strain of Brucella abortus, a strain of a strain of Brucella canis, a strain of a strain of Brucella melitensis, a strain of a strain of Brucella suis, a strain of a strain of Campylobacter jejuni, a strain of Chlamydia pneumoniae, a strain of Chlamydia trachomatis, a strain of Chlamydophila psittaci, a strain of Clostridium botulinum, a strain of Clostridium difficile, a strain of Clostridium perfringens, a strain of Clostridium tetani, a strain of Corynebacterium diphtheria, a strain of Enterobacter sakazakii, a strain of Enterococcus faecalis, a strain of Enterococcus faecium, a strain of Escherichia coli (e.g., E. coli O157 H7), a strain of Francisella tularensis, a strain of Haemophilus influenza, a strain of Helicobacter pylori, a strain of Legionella pneumophila, a strain of Leptospira interrogans, a strain of Listeria monocytogenes, a strain of Mycobacterium leprae, a strain of Mycobacterium tuberculosis, a strain of Mycobacterium ulcerans, a strain of Mycoplasma pneumonia, a strain of Neisseria gonorrhoeae, a strain of Neisseria meningitides, a strain of Pseudomonas aeruginosa, a strain of Rickettsia rickettsia, a strain of Salmonella typhi and Salmonella typhimurium, a strain of Shigella sonnei, a strain of Staphylococcus aureus, a strain of Staphylococcus epidermidis, a strain of Staphylococcus saprophyticus, a strain of Streptococcus agalactiae, a strain of Streptococcus pneumonia, a strain of Streptococcus pyogenes, a strain of Treponema pallidum, a strain of Vibrio cholera, a strain of Yersinia enterocolitica, and, a strain of Yersinia pestis.

[1432] In some embodiments, the bacterium is a commensal bacterium (e.g., a probiotic). In some embodiments, the bacterium has been previously administered to a subject, e.g., as a live biotherapeutic agent. Exemplary commensal bacteria include, but are not limited to, Faecalibacterium prausnitzii (also referred to as Bacteroides praussnitzii), Roseburia hominis, Eubacterium rectale, Dialister invisus, Ruminococcus albus, Ruminococcus gnavus, Ruminococcus torques, Ruminococcus callidus, and Ruminococcus bromii.

[1433] In some embodiments, the analyte is a virus. In some embodiments, the virus is a pathogenic virus. Non-limiting examples of pathogenic viruses belong to the families Adenoviridae, Picornaviridae, Herpesviridae, Hepadnaviridae, Flaviviridae, Retroviridae, Orthomyxoviridae, Paramyxoviridae, Papovaviridae, Polyomavirus, Rhabdoviridae, and Togaviridae.

[1434] In some embodiments, the analyte is a fungus. In some embodiments, the fungi is a pathogenic fungus. Non-limiting examples of pathogenic fungi belong to the genera Asperfillus, Canidia, Cryptococcus, Histoplasma, Pneumocystis, and Stachybotrys. Non-limiting examples of specific pathogenic fungi species include a strain of Aspergillus clavatus, Aspergillus fumigatus, Aspergillus flavus, Canidia albicans, Cryptococcus albidus, Cryptococcus gattii, Cryptococcus laurentii, Cryptococcus neoformans, Histoplasma capsulatum, Pneumocystis jirovecii, Pneumocystis carinii, and Stachybotrys chartarum.

[1435] In some embodiments, the analyte is a protozoan. In some embodiments, the analyte is a pathogenic protozoan. Non-limiting examples of pathogenic protozoa belong to the genera Acanthamoeba, Balamuthia, Cryptosporidium, Dientamoeba, Endolimax, Entamoeba, Giardia, Iodamoeba, Leishmania, Naegleria, Plasmodium, Sappinia, Toxoplasma, Trichomonas, and Trypanosoma. Non-limiting examples of specific pathogenic protozoa species include a strain of Acanthamoeba spp., Balamuthia mandrillaris, Cryptosporidium canis, Cryptosporidium felis, Cryptosporidium hominis, Cryptosporidium meleagridis, Cryptosporidium muris, Cryptosporidium parvum, Dientamoeba fragilis, Endolimax nana, Entamoeba dispar, Entamoeba hartmanni, Entamoeba histolytica, Entamoeba coi, Entamoeba moshkovskii, Giardia lamblia, Iodamoeba butschlii, Leishmania aethiopica, Leishmania braziliensis, Leishmania chagasi, Leishmania donovani, Leishmania infantum, Leishmania major, Leishmania mexicana, Leishmania tropica, Naegleria fowleri, Plasmodium falciparum, Plasmodium knowlesi, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, Sappinia diploidea, Toxoplasma gondii, Trichomonas vaginalis, Trypanosoma brucei, and Trypanosoma cruzi.

[1436] In some embodiments, the analyte is secreted by or expressed on the cell surface of a microorganism (e.g., a bacterium, a colonic bacterium, a viable bacterium, a dead bacterium, a parasite (e.g., Giardia lamblia, Cryptosporidium, Cystoisosporiasis belli, and Balantidium coli), a virus (e.g., a herpes virus, a cytomegalovirus, a herpes simplex virus, an Epstein-Barr virus, a human papilloma virus, a rotavirus, a human herpesvirus-8; Goodgame (1999) Curr. Gastroenterol. Rep. 1(4): 292-300). In some embodiments, the analyte is secreted by or expressed on the cell surface of a Gram-negative bacterium (e.g., E. coli, Helicobacter pylori). In some embodiments, the analyte is secreted by or expressed on the cell surface (e.g., a bacterial surface epitope) of a Gram-positive bacterium (e.g., Staphylococcus aureus, Clostridium botulinum, Clostridium difficile).

[1437] In some embodiments, the analyte is a molecule expressed on the surface of a bacterial cell (e.g., a bacterial cell surface protein). In some embodiments, the analyte is a bacterial toxin (e.g., TcdA and/or TcdB from Clostridium difficile). In some embodiments, the analyte is CFA/I fimbriae, flagella, lipopolysaccharide (LPS), lipoteichoic acid, or a peptidoglycan. Non-limiting examples of bacterium that may express an analyte that can be detected using any of the devices and methods described herein include: Bacillus anthracis, Bacillus cereus, Clostridium botulinum, Clostridium difficile, Escherichia coli, Yersinia pestis, Yersinia enterocolitica, Francisella tularensis, Brucella species, Clostridium perfringens, Burkholderia mallei, Burkholderia pseudomallei, Helicobacter pylori, Staphylococcus species, Mycobacterium species, Group A Streptococcus, Group B Streptococcus, Streptococcus pneumoniae, Francisella tularensis, Salmonella enteritidis, Mycoplasma hominis, Mycoplasma orale, Mycoplasma salivarium, Mycoplasma fermentans, Mycoplasma pneumoniae, Mycobacterium bovis, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium leprae, Rickettsia rickettsii, Rickettsia akari, Rickettsia prowazekii, Rickettsia canada, Bacillus subtilis, Bacillus subtilis niger, Bacillus thuringiensis, Coxiella bumetti, Candida albicans, Bacteroides fragilis, Leptospira interrogans, Listeria monocytogenes, Pasteurella multocida, Salmonella typhi, Salmonella typhimurium, Shigella dysenteriae, Shigella flexneria, Shigella sonnei, Vibrio cholera, and Vibrio parahaemolyticus.

[1438] In some embodiments, the analyte is a byproduct from a bacterium or another microorganism, e.g., helminth ova, enterotoxin (Clostridium difficile toxin A; TcdA), cytotoxin (Clostridium difficile toxin B; TcdB), ammonia. In some embodiments, the analyte is an antigen from a microorganism (e.g., a bacteria, virus, prion, fungus, protozoan or a parasite).

[1439] In some embodiments, the analytes include drugs, metabolites, pesticides, pollutants, and the like. Included among drugs of interest are the alkaloids. Among the alkaloids are morphine alkaloids, which includes morphine, codeine, heroin, dextromethorphan, their derivatives and metabolites; cocaine alkaloids, which include cocaine and benzyl ecgonine, their derivatives and metabolites; ergot alkaloids, which include the diethylamide of lysergic acid; steroid alkaloids; iminazoyl alkaloids; quinazoline alkaloids; isoquinoline alkaloids; quinoline alkaloids, which include quinine and quinidine; diterpene alkaloids, their derivatives and metabolites.

[1440] In some embodiments, the analyte is a steroid selected from the estrogens, androgens, andreocortical steroids, bile acids, cardiotonic glycosides and aglycones, which includes digoxin and digoxigenin, saponins and sapogenins, their derivatives and metabolites. Also included are the steroid mimetic substances, such as diethylstilbestrol.

[1441] In some embodiments, the analyte is a bile acid. In some embodiments, the presence, absence, and/or a specific level of one or more bile acids in the GI tract of a subject is indicative of a condition or disease state (e.g., a GI disorder and/or a non-GI disorder (e.g., a systemic disorder). For example, in some embodiments, the compositions and methods described herein may be used to detect and/or quantify a bile acid in the GI tract of the subject to diagnose a condition such as bile acid malabsorption (also known as bile acid diarrhea). In some embodiments, the analyte is a metabolite in the serotonin, tryptophan and/or kynurenine pathways, including but not limited to, serotonin (5-HT), 5-hydroxyindole acetic acid (5-HIAA), 5-hydroxytryptophan (5-HTP), kynurenine (K), kynurenic acid (KA), 3-hydroxykynurenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), quinolinic acid, anthranilic acid, and combinations thereof. 5-HT is a molecule that plays a role in the regulation of gastrointestinal motility, secretion, and sensation. Imbalances in the levels of 5-HT are associated with several diseases including inflammatory bowel syndrome (IBS), autism, gastric ulcer formation, non-cardiac chest pain, and functional dyspepsia (see, e.g., Faure et al. (2010) Gastroenterology 139(1): 249-58 and Muller et al. (2016) Neuroscience 321: 24-41, and International Publication No. WO 2014/188377, each of which are incorporated herein by reference). Conversion of metabolites within the serotonin, tryptophan and/or kynurenine pathways affects the levels of 5-HT in a subject. Therefore, measuring the levels of one or more of the metabolites in this pathway may be used for the diagnosis, management and treatment of a disease or disorder associated with 5-HT imbalance including but not limited to IBS, autism, carcinoid syndrome, depression, hypertension, Alzheimer's disease, constipation, migraine, and serotonin syndrome. One or more analytes in the serotonin, tryptophan and/or kynurenine pathways can be detected and/or quantitated using, for example, methods and analyte-binding agents that bind to these metabolites including, e.g., antibodies, known in the art (see, e.g., International Publication No. WO2014/188377, the entire contents of which are expressly incorporated herein by reference).

[1442] In some embodiments, the analyte is a lactam having from 5 to 6 annular members selected from barbituates, e.g., phenobarbital and secobarbital, diphenylhydantonin, primidone, ethosuximide, and metabolites thereof.

[1443] In some embodiments, the analyte is an aminoalkylbenzene, with alkyl of from 2 to 3 carbon atoms, selected from the amphetamines; catecholamines, which includes ephedrine, L-dopa, epinephrine; narceine; papaverine; and metabolites thereof.

[1444] In some embodiments, the analyte is a benzheterocyclic selected from oxazepam, chlorpromazine, tegretol, their derivatives and metabolites, the heterocyclic rings being azepines, diazepines and phenothiazines.

[1445] In some embodiments, the analyte is a purine selected from theophylline, caffeine, their metabolites and derivatives.

[1446] In some embodiments, the analyte is marijuana, cannabinol or tetrahydrocannabinol.

[1447] In some embodiments, the analyte is a vitamin such as vitamin A, vitamin B, e.g. vitamin B.sub.12, vitamin C, vitamin D, vitamin E and vitamin K, folic acid, thiamine.

[1448] In some embodiments, the analyte is selected from prostaglandins, which differ by the degree and sites of hydroxylation and unsaturation.

[1449] In some embodiments, the analyte is a tricyclic antidepressant selected from imipramine, dismethylimipramine, amitriptyline, nortriptyline, protriptyline, trimipramine, chlomipramine, doxepine, and desmethyldoxepin.

[1450] In some embodiments, the analyte is selected from anti-neoplastics, including methotrexate.

[1451] In some embodiments, the analyte is an antibiotic as described herein, including, but not limited to, penicillin, chloromycetin, actinomycetin, tetracycline, terramycin, and metabolites and derivatives.

[1452] In some embodiments, the analyte is a nucleoside and nucleotide selected from ATP, NAD, FMN, adenosine, guanosine, thymidine, and cytidine with their appropriate sugar and phosphate substituents.

[1453] In some embodiments, the analyte is selected from methadone, meprobamate, serotonin, meperidine, lidocaine, procainamide, acetylprocainamide, propranolol, griseofulvin, valproic acid, butyrophenones, antihistamines, chloramphenicol, anticholinergic drugs, such as atropine, their metabolites and derivatives.

[1454] In some embodiments, the analyte is a metabolite related to a diseased state. Such metabolites include, but are not limited to spermine, galactose, phenylpyruvic acid, and porphyrin Type 1.

[1455] In some embodiments, the analyte is an aminoglycoside, such as gentamicin, kanamicin, tobramycin, or amikacin.

[1456] In some embodiments, the analyte is a pesticide. Among pesticides of interest are polyhalogenated biphenyls, phosphate esters, thiophosphates, carbamates, polyhalogenated sulfenamides, their metabolites and derivatives.

[1457] In some embodiments, the analyte has a molecular weight of about 500 Da to about 1,000,000 Da (e.g., about 500 to about 500,000 Da, about 1,000 to about 100,000 Da).

[1458] In some embodiments, the analyte is a receptor, with a molecular weight ranging from 10,000 to 210.sup.8 Da, more usually from 10,000 to 10.sup.6 Da. For immunoglobulins, IgA, IgG, IgE and IgM, the molecular weights will generally vary from about 160,000 Da to about 10.sup.6 Da. Enzymes will normally range in molecular weight from about 10,000 Da to about 1,000,000 Da. Natural receptors vary widely, generally having a molecular weight of at least about 25,000 Da and may be 10.sup.6 or higher Da, including such materials as avidin, DNA, RNA, thyroxine binding globulin, thyroxine binding prealbumin, transcortin, etc.

[1459] In some embodiments, the term analyte further includes polynucleotide analytes such as those polynucleotides defined below. These include m-RNA, r-RNA, t-RNA, DNA, DNA-RNA duplexes, etc. The term analyte also includes polynucleotide-binding agents, such as, for example, restriction enzymes, trascription factors, transcription activators, transcription repressors, nucleases, polymerases, histones, DNA repair enzymes, intercalating gagents, chemotherapeutic agents, and the like.

[1460] In some embodiments, the analyte may be a molecule found directly in a sample such as a body fluid from a host. The sample can be examined directly or may be pretreated to render the analyte more readily detectible. Furthermore, the analyte of interest may be determined by detecting an agent probative of the analyte of interest (i.e., an analyte-binding agent), such as a specific binding pair member complementary to the analyte of interest, whose presence will be detected only when the analyte of interest is present in a sample. Thus, the agent probative of the analyte becomes the analyte that is detected in an assay.

[1461] In some embodiments, the analyte a nucleic acid (e.g., a bacterial DNA molecule or a bacterial RNA molecule (e.g., a bacterial tRNA, a transfer-messenger RNA (tmRNA)). See, e.g., Sjostrom et al. (2015) Scientific Reports 5: 15329; Ghosal (2017) Microbial Pathogenesis 104: 161-163; Shen et al. (2012) Cell Host Microbe. 12(4): 509-520.

[1462] In some embodiments, the analyte is a component of an outer membrane vesicle (OMV) (e.g., an OmpU protein, Elluri et al. (2014) PloS One 9: e106731). See, e.g., Kulp and Kuehn (2010) Annual Review of microbiology 64: 163-184; Berleman and Auer (2013) Environmental microbiology 15: 347-354; Wai et al. (1995) Microbiology and immunology 39: 451-456; Lindmark et al. (2009) BMC microbiology 9: 220; Sjostrom et al. (2015) Scientific Reports 5: 15329.

[1463] In some embodiments, the analyte is G-CSF, which can stimulate the bone marrow to produce granulocytes and stem cells and release them into the bloodstream.

[1464] In some embodiments, the analyte is an enzyme such as glutathione S-transferase. For example, the ingestible device can include P28GST, a 28 kDa helminth protein from Schistosoma with potent immunogenic and antioxidant properties. P28GST prevents intestinal inflammation in experimental colitis through a Th2-type response with mucosal eosinophils and can be recombinantly produced (e.g., in S. cerevisiae). See, for example, U.S. Pat. No. 9,593,313, Driss et al., Mucosal Immunology, 2016 9, 322-335; and Capron et al., Gastroenterology, 146(5):S-638.

[1465] In some embodiments, the analyte is a metabolite in the serotonin, tryptophan and/or kynurenine pathways, including but not limited to, serotonin (5-HT), 5-hydroxyindole acetic acid (5-HIAA), 5-hydroxytryptophan (5-HTP), kynurenine (K), kynurenic acid (KA), 3-hydroxykynurenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), quinolinic acid, anthranilic acid, and combinations thereof.

[1466] In some embodiments, analytes are therapeutic agents or drugs. In some embodiments, analytes are biomarkers. The therapeutic agents disclosed herein are can also be analytes. Examples of biomarkers are provided herein.

[1467] In some embodiments, analytes are therapeutic agents, fragments thereof, and metabolites thereof (e.g., antibiotics). In some embodiments, the analytes are antibodies. In some embodiments, the analytes are antibiotics. Additional exemplary analytes (e.g., antibodies and antibiotics) are provided below.

[1468] a. Antibodies

[1469] In some embodiments, the analyte or the analyte-binding agent is an antibody. An antibody is an immunoglobulin molecule capable of specific binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable region of the immunoglobulin molecule. As used herein, the term encompasses not only intact polyclonal or monoclonal antibodies, but also fragments thereof (such as Fab, Fab, F(ab)2, Fv), single chain (ScFv) and domain antibodies), and fusion proteins including an antibody portion, and any other modified configuration of the immunoglobulin molecule that includes an antigen recognition site. The term antibody includes antibody fragments (e.g., antigen-binding fragments) such as an Fv fragment, a Fab fragment, a F(ab)2 fragment, and a Fab fragment. Additional examples of antigen-binding fragments include an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgG1, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgG1, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgA1 or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgA1 or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM). An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant domain of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[1470] As used herein, monoclonal antibody refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies including the population are identical except for possible naturally-occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier monoclonal indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or may be made by recombinant DNA methods such as described in U.S. Pat. No. 4,816,567. The monoclonal antibodies may also be isolated from phage libraries generated using the techniques described in McCafferty et al., 1990, Nature 348:552-554, for example.

[1471] A variable region of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. As known in the art, the variable regions of the heavy and light chain each consist of four framework regions (FR) connected by three complementarity determining regions (CDRs) that contain hypervariable regions. The CDRs in each chain are held together in close proximity by the FRs and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies. There are at least two techniques for determining CDRs: (1) an approach based on cross-species sequence variability (i.e., Kabat et al. Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda Md.)); and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-Lazikani et al, 1997, J. Molec. Biol. 273:927-948). As used herein, a CDR may refer to CDRs defined by either approach or by a combination of both approaches.

[1472] As known in the art, a constant region of an antibody refers to the constant region of the antibody light chain or the constant region of the antibody heavy chain, either alone or in combination.

[1473] A derivative refers to any polypeptide (e.g., an antibody) having a substantially identical amino acid sequence to the naturally occurring polypeptide, in which one or more amino acids have been modified at side groups of the amino acids (e.g., an biotinylated protein or antibody). The term derivative shall also include any polypeptide (e.g., an antibody) which has one or more amino acids deleted from, added to, or substituted from the natural polypeptide sequence, but which retains a substantial amino acid sequence homology to the natural sequence. A substantial sequence homology is any homology greater than 50 percent.

[1474] In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv).sub.2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat. Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO Mol. Med. 9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., J. Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, k-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Pat. No. 5,759,808; Stijlemans et al., J. Biol. Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., J. Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol. 25(2):85-91, 2004), Diabody-CH3 (Guo et al.), Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab)2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther. 8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.

[1475] In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986; WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., J. Exp. Med. 175:217-225, 1992; Kolstelny et al., J. Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Nat. Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., J. Immunol. 152:5368, 1994; Tutt et al., J. Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (FabscFv).sub.2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Pat. No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., J. Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

[1476] In some embodiments, the antibody binds specifically to a metabolite in the serotonin, tryptophan and/or kynurenine pathways, including but not limited to, serotonin (5-HT), 5-hydroxyindole acetic acid (5-HIAA), 5-hydroxytryptophan (5-HTP), kynurenine (K), kynurenic acid (KA), 3-hydroxykynurenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), quinolinic acid, anthranilic acid. Exemplary antibodies that bind to metabolites in these pathways are disclosed, for example, in International Publication No. WO2014/188377, the entire contents of which are incorporated herein by reference.

[1477] In some embodiments, the antibody is specific for a particular genus, species, or strain of a microorganism, and may therefore be used for the detection, analysis and/or quantitation of the microorganism using the detection methods described below. In some embodiments, the antibody specifically binds to a surface-specific biomolecule (e.g., a pilus subunit or a flagella protein) present in a particular genus, species or strain of microorganism, and does not cross-react with other microorganisms. In some embodiments, these antibodies may be used in the methods described herein to diagnose a subject with a particular infection or disease, or to monitor an infection (e.g., during or after treatment). In some embodiments, the antibody specifically binds to an antigen present in a particular genera, species or strain of a microorganism. Exemplary antigens, the corresponding microorganism that can be detected, and the disease caused by the microorganism (in parentheticals) include: outer membrane protein A OmpA (Acinetobacter baumannii, Acinetobacter infections)); HIV p24 antigen, HIV Eenvelope proteins (Gp120, Gp41, Gp160) (HIV (Human immunodeficiency virus), AIDS (Acquired immunodeficiency syndrome)); galactose-inhibitable adherence protein GIAP, 29 kDa antigen Eh29, GaVGaINAc lectin, protein CRT, 125 kDa immunodominant antigen, protein M17, adhesin ADH112, protein STIRP (Entamoeba histolytica, Amoebiasis); protective Antigen PA, edema factor EF, lethal facotor LF, the S-layer homology proteins SLH (Bacillus anthracis, Anthrax); nucleocapsid protein NP, glycoprotein precursor GPC, glycoprotein GP1, glycoprotein GP2 (Junin virus, Argentine hemorrhagic fever); 41 kDa allergen Asp v13, allergen Asp f3, major conidial surface protein rodlet A, protease Pep1p, GPI-anchored protein Gellp, GPI-anchored protein Crflp (Aspergillus genus, Aspergillosis); outer surface protein A OspA, outer surface protein OspB, outer surface protein OspC, decorin binding protein A DbpA, flagellar filament 41 kDa core protein Fla, basic membrane protein A precursor BmpA (Immunodominant antigen P39), outer surface 22 kDa lipoprotein precursor (antigen IPLA7), variable surface lipoprotein vIsE (Borrelia genus, Borrelia infection); OmpA-like transmembrane domain-containing protein Omp31, immunogenic 39-kDa protein M5 P39, 25 kDa outer-membrane immunogenic protein precursor Omp25, outer membrane protein MotY Omp16, conserved outer membrane protein D15, malate dehydrogenase Mdh, component of the Type-IV secretion system (T4SS) VirJ, lipoprotein of unknown function BAB1-0187 (Brucella genus, Brucellosis); major outer membrane protein PorA, flagellin FIaA, surface antigen CjaA, fibronectin binding protein CadF, aspartate/glutamate-binding ABC transporter protein Peb1A, protein FspA1, protein FspA2 (Campylobacter genus, Campylobacteriosis); glycolytic enzyme enolase, secreted aspartyl proteinases SAP1-10, glycophosphatidylinositol (GPI)-linked cell wall protein, adhesin Als3p, cell surface hydrophobicity protein CSH (usually Candida albicans and other Candida species, Candidiasis); envelope glycoproteins (gB, gC, gE, gH, gI, gK, gL) (Varicella zoster virus (VZV), Chickenpox); major outer membrane protein MOMP, probable outer membrane protein PMPC, outer membrane complex protein B OmcB (Chlamydia trachomatis, Chlamydia); major outer membrane protein MOMP, outer membrane protein 2 Omp2, (Chlamydophila pneumoniae, Chlamydophila pneumoniae infection); outer membrane protein U Porin ompU, (Vibrio cholerae, Cholera); surface layer proteins SLPs, Cell Wall Protein CwpV, flagellar protein FliC, flagellar protein FliD (Clostridium difficile, Clostridium difficile infection); acidic ribosomal protein P2 CpP2, mucin antigens Muc1, Muc2, Muc3 Muc4, Muc5, Muc6, Muc7, surface adherence protein CP20, surface adherence protein CP23, surface protein CP12, surface protein CP21, surface protein CP40, surface protein CP60, surface protein CP15, surface-associated glycopeptides gp40, surface-associated glycopeptides gp15, oocyst wall protein AB, profilin PRF, apyrase (Cryptosporidium genus, Cryptosporidiosis); membrane protein pp15, capsid-proximal tegument protein pp150 (Cytomegalovirus, Cytomegalovirus infection); prion protein (vCJD prion, Variant Creutzfeldt-Jakob disease (vCJD, nvCJD)); cyst wall proteins CWP1, CWP2, CWP3, variant surface protein VSP, VSP1, VSP2, VSP3, VSP4, VSP5, VSP6, 56 kDa antigen (Giardia intestinalis, Giardiasis); minor pilin-associated subunit pilC, major pilin subunit and variants pilE, pilS (Neisseria gonorrhoeae, Gonorrhea); outer membrane protein A OmpA, outer membrane protein C OmpC, outer membrane protein K17 OmpK17 (Klebsiella granulomatis, Granuloma inguinale (Donovanosis)); fibronectin-binding protein Sfb (Streptococcus pyogenes, Group A streptococcal infection); outer membrane protein P6 (Haemophilus influenzae, Haemophilus influenzae infection); integral membrane proteins, aggregation-prone proteins, O-antigen, toxin-antigens Stx2B, toxin-antigen Stx1B, adhesion-antigen fragment Int28, protein EspA, protein EspB, Intimin, protein Tir, protein IntC300, protein Eae (Escherichia coli O157:H7, O111 and O104:H4, Hemolytic-uremic syndrome (HUS)); hepatitis A surface antigen HBAg (Hepatitis A Virus, Hepatitis A); hepatitis B surface antigen HBsAg (Hepatitis B Virus, Hepatitis B); envelope glycoprotein E1 gp32 gp35, envelope glycoprotein E2 NS1 gp68 gp70, capsid protein C, (Hepatitis C Virus, Hepatitis C); type IV pilin PilE, outer membrane protein MIP, major outer membrane protein MompS (Legionella pneumophila, Legionellosis (Legionnaires' disease, Pontiac fever)); minor pilin-associated subunit pilC, major pilin subunit and variants pilE, pilS (Neisseria meningitidis, Meningococcal disease); adhesin P1, adhesion P30 (Mycoplasmapneumoniae, Mycoplasma pneumonia); F1 capsule antigen, outer membrane protease Pla, (Yersiniapestis, Plague); surface adhesin PsaA, cell wall surface anchored protein psrP (Streptococcus pneumoniae, Pneumococcal infection); flagellin FliC, invasion protein SipC, glycoprotein gp43, outer membrane protein LamB, outer membrane protein PagC, outer membrane protein TolC, outer membrane protein NmpC, outer membrane protein FadL, transport protein SadA (Salmonella genus, Salmonellosis); collagen adhesin Cna, fibronectin-binding protein A FnbA, secretory antigen SssA (Staphylococcus genus, Staphylococcal food poisoning); collagen adhesin Can (Staphylococcus genus, Staphylococcal infection); fibronectin-binding protein A FbpA (Ag85A), fibronectin-binding protein D FbpD, fibronectin-binding protein C FbpC1, heat-shock protein HSP65, protein PST-S (Mycobacterium tuberculosis, Tuberculosis); and outer membrane protein FobA, outer membrane protein FobB, type IV pili glycosylation protein, outer membrane protein tolC, protein TolQ (Francisella tularensis, Tularemia). Additional exemplary microorganisms and corresponding antigens are disclosed, e.g., in U.S. Publication No. 2015/0118264, the entire contents of which are expressly incorporated herein by reference.

[1478] In some embodiments, a plurality of antibodies (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, or more antibodies) are used as analyte-binding agents in any of the methods described herein (e.g., to detect the presence of one or more analytes in a sample). In some embodiments, the plurality of antibodies bind to the same analyte (e.g., an antigen). In some embodiments, the plurality of antibodes bind to the same epitope present on the analyte (e.g., an antigen). In some embodiments, the plurality of antibodies bind to different epitopes present on the same analyte. In some embodiments, the plurality of antibodies bind to overlapping epitopes present on the same analyte. In some embodiments, the plurality of antibodies bind to non-overlapping epitopes present on the same analyte.

Antibiotics

[1479] In some embodiments, the analyte or analyte-binding agent is an antibiotic. An antibiotic or antibiotic agent refers to a substance that has the capacity to inhibit or slow down the growth of, or to destroy bacteria and/or other microorganisms. In some embodiments, the antibiotic agent is a bacteriostatic antibiotic agent. In some embodiments, the antibiotic is a bacteriolytic antibiotic agent. Exemplary antibiotic agents are set forth in the U.S. Patent Publication US 2006/0269485, which is hereby incorporated by reference herein in its entirety.

[1480] In some embodiments, the antibiotic agent is selected from the classes consisting of beta-lactam antibiotics, aminoglycosides, ansa-type antibiotics, anthraquinones, antibiotic azoles, antibiotic glycopeptides, macrolides, antibiotic nucleosides, antibiotic peptides, antibiotic polyenes, antibiotic polyethers, quinolones, antibiotic steroids, sulfonamides, tetracycline, dicarboxylic acids, antibiotic metals, oxidizing agents, substances that release free radicals and/or active oxygen, cationic antimicrobial agents, quaternary ammonium compounds, biguanides, triguanides, bisbiguanides and analogs and polymers thereof and naturally occurring antibiotic compounds. In some embodiments, the antibiotic is rifaximin.

[1481] Beta-lactam antibiotics include, but are not limited to, 2-(3-alanyl)clavam, 2-hydroxymethylclavam, 8-epi-thienamycin, acetyl-thienamycin, amoxicillin, amoxicillin sodium, amoxicillin trihydrate, amoxicillin-potassium clavulanate combination, ampicillin, ampicillin sodium, ampicillin trihydrate, ampicillin-sulbactam, apalcillin, aspoxicillin, azidocillin, azlocillin, aztreonam, bacampicillin, biapenem, carbenicillin, carbenicillin disodium, carfecillin, carindacillin, carpetimycin, cefacetril, cefaclor, cefadroxil, cefalexin, cefaloridine, cefalotin, cefamandole, cefamandole, cefapirin, cefatrizine, cefatrizine propylene glycol, cefazedone, cefazolin, cefbuperazone, cefcapene, cefcapene pivoxil hydrochloride, cefdinir, cefditoren, cefditoren pivoxil, cefepime, cefetamet, cefetamet pivoxil, cefixime, cefinenoxime, cefinetazole, cefminox, cefminox, cefmolexin, cefodizime, cefonicid, cefoperazone, ceforanide, cefoselis, cefotaxime, cefotetan, cefotiam, cefoxitin, cefozopran, cefpiramide, cefpirome, cefpodoxime, cefpodoxime proxetil, cefprozil, cefquinome, cefradine, cefroxadine, cefsulodin, ceftazidime, cefteram, cefteram pivoxil, ceftezole, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime, cefuroxime axetil, cephalosporin, cephamycin, chitinovorin, ciclacillin, clavulanic acid, clometocillin, cloxacillin, cycloserine, deoxy pluracidomycin, dicloxacillin, dihydro pluracidomycin, epicillin, epithienamycin, ertapenem, faropenem, flomoxef, flucloxacillin, hetacillin, imipenem, lenampicillin, loracarbef, mecillinam, meropenem, metampicillin, meticillin, mezlocillin, moxalactam, nafcillin, northienamycin, oxacillin, panipenem, penamecillin, penicillin, phenethicillin, piperacillin, tazobactam, pivampicillin, pivcefalexin, pivmecillinam, pivmecillinam hydrochloride, pluracidomycin, propicillin, sarmoxicillin, sulbactam, sulbenicillin, talampicillin, temocillin, terconazole, thienamycin, ticarcillin and analogs, salts and derivatives thereof.

[1482] Aminoglycosides include, but are not limited to, 1,2-N-DL-isoseryl-3,4-dideoxykanamycin B, 1,2-N-DL-isoseryl-kanamycin B, 1,2-N[(S)-4-amino-2-hydroxybutyryl]-3,4-dideoxykanamycin B, 1,2-N[(S)-4-amino-2-hydroxybutyryl]-kanamycin B, 1-N-(2-Aminobutanesulfonyl) kanamycin A, 1-N-(2-aminoethanesulfonyl)3,4-dideoxyribostamycin, 1-N-(2-Aminoethanesulfonyl)3-deoxyribostamycin, 1-N-(2-aminoethanesulfonyl)34-dideoxykanamycin B, 1-N-(2-aminoethanesulfonyl)kanamycin A, 1-N-(2-aminoethanesulfonyl)kanamycin B, 1-N-(2-aminoethanesulfonyl)ribostamycin, 1-N-(2-aminopropanesulfonyl)3-deoxykanamycin B, 1-N-(2-aminopropanesulfonyl)34-dideoxykanamycin B, 1-N-(2-aminopropanesulfonyl)kanamycin A, 1-N-(2-aminopropanesulfonyl)kanamycin B, 1-N-(L-4-amino-2-hydroxy-butyryl)2,3-dideoxy-2-fluorokanamycin A, 1-N-(L-4-amino-2-hydroxy-propionyl)2,3-dideoxy-2-fluorokanamycin A, 1-N-DL-3,4-dideoxy-isoserylkanamycin B, 1-N-DL-isoserylkanamycin, 1-N-DL-isoserylkanamycin B, 1-N-[L-()-(alpha-hydroxy-gamma-aminobutyryl)]-K-62-2,2,3-dideoxy-2-fluorokanamycin A,2-hydroxygentamycin A3,2-hydroxygentamycin B, 2-hydroxygentamycin B1, 2-hydroxygentamycin JI-20A, 2-hydroxygentamycin JI-20B, 3-N-methyl-4-C-methyl-3,4-dodeoxy kanamycin A, 3-N-methyl-4-C-methyl-3,4-dodeoxy kanamycin B, 3-N-methyl-4-C-methyl-3,4-dodeoxy-6-methyl kanamycin B, 3,4-Dideoxy-3-eno-ribostamycin,3,4-dideoxyneamine,3,4-dideoxyribostamycin, 3-deoxy-6-N-methyl-kanamycin B,3-deoxyneamine,3-deoxyribostamycin, 3-oxysaccharocin,3,3-nepotrehalosadiamine, 3-demethoxy-2-N-formimidoylistamycin B disulfate tetrahydrate, 3-demethoxyistamycin B,3-O-demethyl-2-N-formimidoylistamycin B, 3-O-demethylistamycin B,3-trehalosamine,4,6-dideoxydibekacin, 4-N-glycyl-KA-6606VI, 538-Amino-3,4,5-trideoxy-butirosin A, 6-deoxydibekacin,6-epifortimicin A, 6-deoxy-neomycin (structure 6-deoxy-neomycin B),6-deoxy-neomycin B, 6-deoxy-neomycin C, 6-deoxy-paromomycin, acmimycin, AHB-3,4-dideoxyribostamycin, AHB-3-deoxykanamycin B, AHB-3-deoxyneamine, AHB-3-deoxyribostamycin, AHB-4-6-dideoxydibekacin, AHB-6-deoxydibekacin, AHB-dideoxyneamine, AHB-kanamycin B, AHB-methyl-3-deoxykanamycin B, amikacin, amikacin sulfate, apramycin, arbekacin, astromicin, astromicin sulfate, bekanamycin, bluensomycin, boholmycin, butirosin, butirosin B, catenulin, coumamidine gamma1, coumamidine gamma 2,D,L-1-N-(alpha-hydroxy-beta-aminopropionyl)-XK-62-2, dactimicin, de-O-methyl-4-N-glycyl-KA-6606VI, de-O-methyl-KA-6606I, de-O-methyl-KA-7038I, destomycin A, destomycin B, di-N6,O3-demethylistamycin A, dibekacin, dibekacin sulfate, dihydrostreptomycin, dihydrostreptomycin sulfate, epi-formamidoylglycidylfortimicin B, epihygromycin, formimidoyl-istamycin A, formimidoyl-istamycin B, fortimicin B, fortimicin C, fortimicin D, fortimicin KE, fortimicin KF, fortimicin KG, fortimicin KG1 (stereoisomer KG1/KG2), fortimicin KG2 (stereoisomer KG1/KG2), fortimicin KG3, framycetin, framycetin sulphate, gentamicin, gentamycin sulfate, globeomycin, hybrimycin A1, hybrimycin A2, hybrimycin B1, hybrimycin B2, hybrimycin C1, hybrimycin C2, hydroxystreptomycin, hygromycin, hygromycin B, isepamicin, isepamicin sulfate, istamycin, kanamycin, kanamycin sulphate, kasugamycin, lividomycin, marcomycin, micronomicin, micronomicin sulfate, mutamicin, myomycin, N-demethyl-7-O-demethylcelesticetin, demethylcelesticetin, methanesulfonic acid derivative of istamycin, nebramycin, nebramycin, neomycin, netilmicin, oligostatin, paromomycin, quintomycin, ribostamycin, saccharocin, seldomycin, sisomicin, sorbistin, spectinomycin, streptomycin, tobramycin, trehalosmaine, trestatin, validamycin, verdamycin, xylostasin, zygomycin and analogs, salts and derivatives thereof.

[1483] Ansa-type antibiotics include, but are not limited to, 21-hydroxy-25-demethyl-25-methylth ioprotostreptovaricin, 3-methylth iorifamycin, ansamitocin, atropisostreptovaricin, awamycin, halomicin, maytansine, naphthomycin, rifabutin, rifamide, rifampicin, rifamycin, rifapentine, rifaximin (e.g., Xifaxan), rubradirin, streptovaricin, tolypomycin and analogs, salts and derivatives thereof.

[1484] Antibiotic anthraquinones include, but are not limited to, auramycin, cinerubin, ditrisarubicin, ditrisarubicin C, figaroic acid fragilomycin, minomycin, rabelomycin, rudolfomycin, sulfurmycin and analogs, salts and derivatives thereof.

[1485] Antibiotic azoles include, but are not limited to, azanidazole, bifonazole, butoconazol, chlormidazole, chlormidazole hydrochloride, cloconazole, cloconazole monohydrochloride, clotrimazol, dimetridazole, econazole, econazole nitrate, enilconazole, fenticonazole, fenticonazole nitrate, fezatione, fluconazole, flutrimazole, isoconazole, isoconazole nitrate, itraconazole, ketoconazole, lanoconazole, metronidazole, metronidazole benzoate, miconazole, miconazole nitrate, neticonazole, nimorazole, niridazole, omoconazol, ornidazole, oxiconazole, oxiconazole nitrate, propenidazole, secnidazol, sertaconazole, sertaconazole nitrate, sulconazole, sulconazole nitrate, tinidazole, tioconazole, voriconazol and analogs, salts and derivatives thereof.

[1486] Antibiotic glycopeptides include, but are not limited to, acanthomycin, actaplanin, avoparcin, balhimycin, bleomycin B (copper bleomycin), chloroorienticin, chloropolysporin, demethylvancomycin, enduracidin, galacardin, guanidylfungin, hachimycin, demethylvancomycin, N-nonanoyl-teicoplanin, phleomycin, platomycin, ristocetin, staphylocidin, talisomycin, teicoplanin, vancomycin, victomycin, xylocandin, zorbamycin and analogs, salts and derivatives thereof.

[1487] Macrolides include, but are not limited to, acetylleucomycin, acetylkitasamycin, angolamycin, azithromycin, bafilomycin, brefeldin, carbomycin, chalcomycin, cirramycin, clarithromycin, concanamycin, deisovaleryl-niddamycin, demycinosyl-mycinamycin, Di-O-methyltiacumicidin, dirithromycin, erythromycin, erythromycin estolate, erythromycin ethyl succinate, erythromycin lactobionate, erythromycin stearate, flurithromycin, focusin, foromacidin, haterumalide, haterumalide, josamycin, josamycin ropionate, juvenimycin, juvenimycin, kitasamycin, ketotiacumicin, lankavacidin, lankavamycin, leucomycin, machecin, maridomycin, megalomicin, methylleucomycin, methymycin, midecamycin, miocamycin, mycaminosyltylactone, mycinomycin, neutramycin, niddamycin, nonactin, oleandomycin, phenylacetyideltamycin, pamamycin, picromycin, rokitamycin, rosaramicin, roxithromycin, sedecamycin, shincomycin, spiramycin, swalpamycin, tacrolimus, telithromycin, tiacumicin, tilmicosin, treponemycin, troleandomycin, tylosin, venturicidin and analogs, salts and derivatives thereof.

[1488] Antibiotic nucleosides include, but are not limited to, amicetin, angustmycin, azathymidine, blasticidin S, epiroprim, flucytosine, gougerotin, mildiomycin, nikkomycin, nucleocidin, oxanosine, oxanosine, puromycin, pyrazomycin, showdomycin, sinefungin, sparsogenin, spicamycin, tunicamycin, uracil polyoxin, vengicide and analogs, salts and derivatives thereof.

[1489] Antibiotic peptides include, but are not limited to, actinomycin, aculeacin, alazopeptin, amfomycin, amythiamycin, antifungal from Zalerion arboricola, antrimycin, apid, apidaecin, aspartocin, auromomycin, bacileucin, bacillomycin, bacillopeptin, bacitracin, bagacidin, beminamycin, beta-alanyl-L-tyrosine, bottromycin, capreomycin, caspofungine, cepacidine, cerexin, cilofungin, circulin, colistin, cyclodepsipeptide, cytophagin, dactinomycin, daptomycin, decapeptide, desoxymulundocandin, echanomycin, echinocandin B, echinomycin, ecomycin, enniatin, etamycin, fabatin, ferrimycin, ferrimycin, ficellomycin, fluoronocathiacin, fusaricidin, gardimycin, gatavalin, globopeptin, glyphomycin, gramicidin, herbicolin, iomycin, iturin, iyomycin, izupeptin, janiemycin, janthinocin, jolipeptin, katanosin, killertoxin, lipopeptide antibiotic, lipopeptide from Zalerion sp., lysobactin, lysozyme, macromomycin, magainin, melittin, mersacidin, mikamycin, mureidomycin, mycoplanecin, mycosubtilin, neopeptifluorin, neoviridogrisein, netropsin, nisin, nocathiacin, nocathiacin 6-deoxyglycoside, nosiheptide, octapeptin, pacidamycin, pentadecapeptide, peptifluorin, permetin, phytoactin, phytostreptin, planothiocin, plusbacin, polcillin, polymyxin antibiotic complex, polymyxin B, polymyxin B1, polymyxin F, preneocarzinostatin, quinomycin, quinupristin-dalfopristin, safracin, salmycin, salmycin, salmycin, sandramycin, saramycetin, siomycin, sperabillin, sporamycin, a Streptomyces compound, subtilin, teicoplanin aglycone, telomycin, thermothiocin, thiopeptin, thiostrepton, tridecaptin, tsushimycin, tuberactinomycin, tuberactinomycin, tyrothricin, valinomycin, viomycin, virginiamycin, zervacin and analogs, salts and derivatives thereof.

[1490] In some embodiments, the antibiotic peptide is a naturally-occurring peptide that possesses an antibacterial and/or an antifungal activity. Such peptide can be obtained from an herbal or a vertebrate source.

[1491] Polyenes include, but are not limited to, amphotericin, amphotericin, aureofungin, ayfactin, azalomycin, blasticidin, candicidin, candicidin methyl ester, candimycin, candimycin methyl ester, chinopricin, filipin, flavofungin, fradicin, hamycin, hydropricin, levorin, lucensomycin, lucknomycin, mediocidin, mediocidin methyl ester, mepartricin, methylamphotericin, natamycin, niphimycin, nystatin, nystatin methyl ester, oxypricin, partricin, pentamycin, perimycin, pimaricin, primycin, proticin, rimocidin, sistomycosin, sorangicin, trichomycin and analogs, salts and derivatives thereof.

[1492] Polyethers include, but are not limited to, 20-deoxy-epi-narasin, 20-deoxysalinomycin, carriomycin, dianemycin, dihydrolonomycin, etheromycin, ionomycin, iso-lasalocid, lasalocid, lenoremycin, lonomycin, lysocellin, monensin, narasin, oxolonomycin, a polycyclic ether antibiotic, salinomycin and analogs, salts and derivatives thereof.

[1493] Quinolones include, but are not limited to, an alkyl-methylendioxy-4(1H)-oxocinnoline-3-carboxylic acid, alatrofloxacin, cinoxacin, ciprofloxacin, ciprofloxacin hydrochloride, danofloxacin, dermofongin A, enoxacin, enrofloxacin, fleroxacin, flumequine, gatifloxacin, gemifloxacin, grepafloxacin, levofloxacin, lomefloxacin, lomefloxacin, hydrochloride, miloxacin, moxifloxacin, nadifloxacin, nalidixic acid, nifuroquine, norfloxacin, ofloxacin, orbifloxacin, oxolinic acid, pazufloxacine, pefloxacin, pefloxacin mesylate, pipemidic acid, piromidic acid, premafloxacin, rosoxacin, rufloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin and analogs, salts and derivatives thereof.

[1494] Antibiotic steroids include, but are not limited to, aminosterol, ascosteroside, cladosporide A, dihydrofusidic acid, dehydro-dihydrofusidic acid, dehydrofusidic acid, fusidic acid, squalamine and analogs, salts and derivatives thereof.

[1495] Sulfonamides include, but are not limited to, chloramine, dapsone, mafenide, phthalylsulfathiazole, succinylsulfathiazole, sulfabenzamide, sulfacetamide, sulfachlorpyridazine, sulfadiazine, sulfadiazine silver, sulfadicramide, sulfadimethoxine, sulfadoxine, sulfaguanidine, sulfalene, sulfamazone, sulfamerazine, sulfamethazine, sulfamethizole, sulfamethoxazole, sulfamethoxypyridazine, sulfamonomethoxine, sulfamoxol, sulfanilamide, sulfaperine, sulfaphenazol, sulfapyridine, sulfaquinoxaline, sulfasuccinamide, sulfathiazole, sulfathiourea, sulfatolamide, sulfatriazin, sulfisomidine, sulfisoxazole, sulfisoxazole acetyl, sulfacarbamide and analogs, salts and derivatives thereof.

[1496] Tetracyclines include, but are not limited to, dihydrosteffimycin, demethyltetracycline, aclacinomycin, akrobomycin, baumycin, bromotetracycline, cetocyclin, chlortetracycline, clomocycline, daunorubicin, demeclocycline, doxorubicin, doxorubicin hydrochloride, doxycycline, lymecyclin, marcellomycin, meclocycline, meclocycline sulfosalicylate, methacycline, minocycline, minocycline hydrochloride, musettamycin, oxytetracycline, rhodirubin, rolitetracycline, rubomycin, serirubicin, steffimycin, tetracycline and analogs, salts and derivatives thereof.

[1497] Dicarboxylic acids, having between about 6 and about 14 carbon atoms in their carbon atom skeleton are particularly useful in the treatment of disorders of the skin and mucosal membranes that involve microbial. Suitable dicarboxylic acid moieties include, but are not limited to, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, 1,11-undecanedioic acid, 1,12-dodecanedioic acid, 1,13-tridecanedioic acid and 1,14-tetradecanedioic acid. Thus, in one or more embodiments of the present disclosure, dicarboxylic acids, having between about 6 and about 14 carbon atoms in their carbon atom skeleton, as well as their salts and derivatives (e.g., esters, amides, mercapto-derivatives, anhydraides), are useful immunomodulators in the treatment of disorders of the skin and mucosal membranes that involve inflammation. Azelaic acid and its salts and derivatives are preferred. It has antibacterial effects on both aerobic and anaerobic organisms, particularly Propionibacterium acnes and Staphylococcus epidermidis, normalizes keratinization, and has a cytotoxic effect on malignant or hyperactive melanocytes. In a preferred embodiment, the dicarboxylic acid is azelaic acid in a concentration greater than 10%. Preferably, the concentration of azelaic acid is between about 10% and about 25%. In such concentrates, azelaic acid is suitable for the treatment of a variety of skin disorders, such as acne, rosacea and hyperpigmentation.

[1498] In some embodiments, the antibiotic agent is an antibiotic metal. A number of metals ions have been shown to possess antibiotic activity, including silver, copper, zinc, mercury, tin, lead, bismutin, cadmium, chromium and ions thereof. It has been theorized that these antibiotic metal ions exert their effects by disrupting respiration and electron transport systems upon absorption into bacterial or fungal cells. Anti-microbial metal ions of silver, copper, zinc, and gold, in particular, are considered safe for in vivo use. Anti-microbial silver and silver ions are particularly useful due to the fact that they are not substantially absorbed into the body. Thus, in one or more embodiment, the antibiotic metal consists of an elemental metal, selected from the group consisting of silver, copper, zinc, mercury, tin, lead, bismutin, cadmium, chromium and gold, which is suspended in the composition as particles, microparticles, nanoparticles or colloidal particles. The antibiotic metal can further be intercalated in a chelating substrate.

[1499] In further embodiments, the antibiotic metal is ionic. The ionic antibiotic metal can be presented as an inorganic or organic salt (coupled with a counterion), an organometallic complex or an intercalate. Non-binding examples of counter inorganic and organic ions are sulfadiazine, acetate, benzoate, carbonate, iodate, iodide, lactate, laurate, nitrate, oxide, and palmitate, a negatively charged protein. In preferred embodiments, the antibiotic metal salt is a silver salt, such as silver acetate, silver benzoate, silver carbonate, silver iodate, silver iodide, silver lactate, silver laurate, silver nitrate, silver oxide, silver palmitate, silver protein, and silver sulfadiazine.

[1500] In one or more embodiments, the antibiotic metal or metal ion is embedded into a substrate, such as a polymer, or a mineral (such as zeolite, clay and silica).

[1501] In one or more embodiments, the antibiotic agent includes strong oxidants and free radical liberating compounds, such as oxygen, hydrogen peroxide, benzoyl peroxide, elemental halogen species, as well as oxygenated halogen species, bleaching agents (e.g., sodium, calcium or magnesium hypochloride and the like), perchlorite species, iodine, iodate, and benzoyl peroxide. Organic oxidizing agents, such as quinones, are also included. Such agents possess a potent broad-spectrum activity.

[1502] In one or more embodiments, the antibiotic agent is a cationic antimicrobial agent. The outermost surface of bacterial cells universally carries a net negative charge, making them sensitive to cationic substances. Examples of cationic antibiotic agents include: quaternary ammonium compounds (QAC's)-QAC's are surfactants, generally containing one quaternary nitrogen associated with at least one major hydrophobic moiety; alkyltrimethyl ammonium bromides are mixtures of where the alkyl group is between 8 and 18 carbons long, such as cetrimide (tetradecyltrimethylammonium bromide); benzalkonium chloride, which is a mixture of n-alkyldimethylbenzyl ammonium chloride where the alkyl groups (the hydrophobic moiety) can be of variable length; dialkylmethyl ammonium halides; dialkylbenzyl ammonium halides; and QAC dimmers, which bear bi-polar positive charges in conjunction with interstitial hydrophobic regions.

[1503] In one or more embodiments, the cationic antimicrobial agent is a polymer. Cationic antimicrobial polymers include, for example, guanide polymers, biguanide polymers, or polymers having side chains containing biguanide moieties or other cationic functional groups, such as benzalkonium groups or quarternium groups (e.g., quaternary amine groups). It is understood that the term polymer as used herein includes any organic material including three or more repeating units, and includes oligomers, polymers, copolymers, block copolymers, terpolymers, etc. The polymer backbone may be, for example a polyethylene, ploypropylene or polysilane polymer.

[1504] In one or more embodiments, the cationic antimicrobial polymer is a polymeric biguanide compound. When applied to a substrate, such a polymer is known to form a barrier film that can engage and disrupt a microorganism. An exemplary polymeric biguanide compound is polyhexamethylene biguanide (PHMB) salts. Other exemplary biguanide polymers include, but are not limited to poly(hexamethylenebiguanide), poly(hexamethylenebiguanide) hydrochloride, poly(hexamethylenebiguanide) gluconate, poly(hexamethylenebiguanide) stearate, or a derivative thereof. In one or more embodiments, the antimicrobial material is substantially water-insoluble.

[1505] In some embodiments, the antibiotic agent is selected from the group of biguanides, triguanides, bisbiguanides and analogs thereof.

[1506] Guanides, biguanides, biguanidines and triguanides are unsaturated nitrogen containing molecules that readily obtain one or more positive charges, which make them effective antimicrobial agents. The basic structures a guanide, a biguanide, a biguanidine and a triguanide are provided below.

##STR00115##

In some embodiments, the guanide, biguanide, biguanidine or triguanide, provide bi-polar configurations of cationic and hydrophobic domains within a single molecule.

[1507] Examples of guanides, biguanides, biguanidines and triguanides that are currently been used as antibacterial agents include chlorhexidine and chlorohexidine salts, analogs and derivatives, such as chlorhexidine acetate, chlorhexidine gluconate and chlorhexidine hydrochloride, picloxydine, alexidine and polihexanide. Other examples of guanides, biguanides, biguanidines and triguanides that can conceivably be used according to the present disclosure are chlorproguanil hydrochloride, proguanil hydrochloride (currently used as antimalarial agents), mefformin hydrochloride, phenformin and buformin hydrochloride (currently used as antidiabetic agents).

[1508] Yet, in one or more embodiments, the antibiotic is a non-classified antibiotic agent, including, without limitation, aabomycin, acetomycin, acetoxycycloheximide, acetylnanaomycin, an Actinoplanes sp. compound, actinopyrone, aflastatin, albacarcin, albacarcin, albofungin, albofungin, alisamycin, alpha-R,S-methoxycarbonylbenzylmonate, altromycin, amicetin, amycin, amycin demanoyl compound, amycine, amycomycin, anandimycin, anisomycin, anthramycin, anti-syphilis immune substance, anti-tuberculosis immune substance, an antibiotic from Escherichia coli, an antibiotic from Streptomyces refuineus, anticapsin, antimycin, aplasmomycin, aranorosin, aranorosinol, arugomycin, ascofuranone, ascomycin, ascosin, Aspergillusflavus antibiotic, asukamycin, aurantinin, an Aureolic acid antibiotic substance, aurodox, avilamycin, azidamfenicol, azidimycin, bacillaene, a Bacillus larvae antibiotic, bactobolin, benanomycin, benzanthrin, benzylmonate, bicozamycin, bravomicin, brodimoprim, butalactin, calcimycin, calvatic acid, candiplanecin, carumonam, carzinophilin, celesticetin, cepacin, cerulenin, cervinomycin, chartreusin, chloramphenicol, chloramphenicol palmitate, chloramphenicol succinate sodium, chlorflavonin, chlorobiocin, chlorocarcin, chromomycin, ciclopirox, ciclopirox olamine, citreamicin, cladosporin, clazamycin, clecarmycin, clindamycin, coliformin, collinomycin, copiamycin, corallopyronin, corynecandin, coumermycin, culpin, cuprimyxin, cyclamidomycin, cycloheximide, dactylomycin, danomycin, danubomycin, delaminomycin, demethoxyrapamycin, demethylscytophycin, dermadin, desdamethine, dexylosyl-benanomycin, pseudoaglycone, dihydromocimycin, dihydronancimycin, diumycin, dnacin, dorrigocin, dynemycin, dynemycin triacetate, ecteinascidin, efrotomycin, endomycin, ensanchomycin, equisetin, ericamycin, esperamicin, ethylmonate, everninomicin, feldamycin, flambamycin, flavensomycin, florfenicol, fluvomycin, fosfomycin, fosfonochlorin, fredericamycin, frenolicin, fumagillin, fumifungin, funginon, fusacandin, fusafungin, gelbecidine, glidobactin, grahamimycin, granaticin, griseofulvin, griseoviridin, grisonomycin, hayumicin, hayumicin, hazymicin, hedamycin, heneicomycin, heptelicid acid, holomycin, humidin, isohematinic acid, karnatakin, kazusamycin, kristenin, L-dihydrophenylalanine, a L-isoleucyl-L-2-amino-4-(4-amino-2,5-cyclohexadienyl) derivative, lanomycin, leinamycin, leptomycin, libanomycin, lincomycin, lomofungin, lysolipin, magnesidin, manumycin, melanomycin, methoxycarbonylmethylmonate, methoxycarbonylethylmonate, methoxycarbonylphenylmonate, methyl pseudomonate, methylmonate, microcin, mitomalcin, mocimycin, moenomycin, monoacetyl cladosporin, monomethyl cladosporin, mupirocin, mupirocin calcium, mycobacidin, myriocin, myxopyronin, pseudoaglycone, nanaomycin, nancimycin, nargenicin, neocarcinostatin, neoenactin, neothramycin, nifurtoinol, nocardicin, nogalamycin, novobiocin, octylmonate, olivomycin, orthosomycin, oudemansin, oxirapentyn, oxoglaucine methiodide, pactacin, pactamycin, papulacandin, paulomycin, phaeoramularia fungicide, phenelfamycin, phenyl, cerulenin, phenylmonate, pholipomycin, pirlimycin, pleuromutilin, a polylactone derivative, polynitroxin, polyoxin, porfiromycin, pradimicin, prenomycin, prop-2-enylmonate, protomycin, Pseudomonas antibiotic, pseudomonic acid, purpuromycin, pyrinodemin, pyrrolnitrin, pyrrolomycin, amino, chloro pentenedioic acid, rapamycin, rebeccamycin, resistomycin, reuterin, reveromycin, rhizocticin, roridin, rubiflavin, naphthyridinomycin, saframycin, saphenamycin, sarkomycin, sarkomycin, sclopularin, selenomycin, siccanin, spartanamicin, spectinomycin, spongistatin, stravidin, streptolydigin, Streptomyces arenae antibiotic complex, streptonigrin, streptothricins, streptovitacin, streptozotocine, a strobilurin derivative, stubomycin, sulfamethoxazol-trimethoprim, sakamycin, tejeramycin, terpentecin, tetrocarcin, thermorubin, thermozymocidin, thiamphenicol, thioaurin, thiolutin, thiomarinol, thiomarinol, tirandamycin, tolytoxin, trichodermin, trienomycin, trimethoprim, trioxacarcin, tyrissamycin, umbrinomycin, unphenelfamycin, urauchimycin, usnic acid, uredolysin, variotin, vermisporin, verrucarin and analogs, salts and derivatives thereof.

[1509] In one or more embodiments, the antibiotic agent is a naturally occurring antibiotic compound. As used herein, the term naturally-occurring antibiotic agent includes all antibiotics that are obtained, derived or extracted from plant or vertebrate sources. Non-limiting examples of families of naturally-occurring antibiotic agents include phenol, resorcinol, antibiotic aminoglycosides, anamycin, quinines, anthraquinones, antibiotic glycopeptides, azoles, macrolides, avilamycin, agropyrene, cnicin, aucubin antibioticsaponin fractions, berberine (isoquinoline alkaloid), arctiopicrin (sesquiterpene lactone), lupulone, humulone (bitter acids), allicin, hyperforin, echinacoside, coniosetin, tetramic acid, imanine and novoimanine.

[1510] Ciclopirox and ciclopiroxolamine possess fungicidal, fungistatic and sporicidal activity. They are active against a broad spectrum of dermatophytes, yeasts, moulds and other fungi, such as Trichophytons species, Microsporum species, Epidermophyton species and yeasts (Candida albicans, Candida glabrata, other candida species and Cryptococcus neoformans). Some Aspergillus species are sensitive to ciclopirox as are some Penicillium. Likewise, ciclopirox is effective against many Gram-positive and Gram-negative bacteria (e.g., Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus and Streptococcus species), as well as Mycoplasma species, Trichomonas vaginalis and Actinomyces.

[1511] Plant oils and extracts which contain antibiotic agents are also useful. Non-limiting examples of plants that contain agents include thyme, Perilla, lavender, tea tree, Terfezia clayeryi, Micromonospora, Putterlickia verrucosa, Putterlickia pyracantha, Putterlickia retrospinosa, Maytenus ilicifolia, Maytenus evonymoides, Maytenus aquifolia, Faenia interjecta, Cordyceps sinensis, couchgrass, holy thistle, plantain, burdock, hops, echinacea, buchu, chaparral, myrrh, red clover and yellow dock, garlic, and St. John's wort. Mixtures of the antibiotic agents as described herein may also be employed.

Combination Detection:

[1512] Any combination of the analytes disclosed herein can be detected using any of the methods described herein. In particular, any combination disclosed herein can be detected using any of the methods described herein.

[1513] A photosensitizer as used herein refers to a sensitizer for generation of singlet oxygen usually by excitation with light. Exemplary photosensitizers suitable for use include those described in U.S. Pat. Nos. 6,251,581, 5,516,636, 8,907,081, 6,545,012, 6,331,530, 8,247,180, 5,763,602, 5,705,622, 5,516,636, 7,217,531, and U.S. Patent Publication No. 2007/0059316, all of which are herein expressly incorporated by reference in their entireties. The photosensitizer can be photoactivatable (e.g., dyes and aromatic compounds) or chemiactivated (e.g., enzymes and metal salts). When excited by light the photosensitizer is usually a compound comprised of covalently bonded atoms, usually with multiple conjugated double or triple bonds. The compound should absorb light in the wavelength range of 200-1100 nm, usually 300-1000 nm, e.g., 450-950 nm, with an extinction coefficient at its absorbance maximum greater than 500 M.sup.1 cm.sup.1, e.g., at least 5000 M.sup.1 cm.sup.1, or at least 50,000 M.sup.1 cm.sup.1 at the excitation wavelength. The lifetime of an excited state produced following absorption of light in the absence of oxygen will usually be at least 100 nsec, e.g., at least 1 sec. In general, the lifetime must be sufficiently long to permit energy transfer to oxygen, which will normally be present at concentrations in the range of 10.sup.5 to 10.sup.31 3M depending on the medium. The sensitizer excited state will usually have a different spin quantum number (S) than its ground state and will usually be a triplet (S=1) when, as is usually the case, the ground state is a singlet (SO). In some embodiments, the sensitizer will have a high intersystem crossing yield. That is, photoexcitation of a sensitizer will produce the long lived state (usually triplet) with an efficiency of at least 10%, at least 40%, e.g., greater than 80%. The photosensitizer will usually be at most weakly fluorescent under the assay conditions (quantum yield usually less that 0.5, or less that 0.1).

[1514] Photosensitizers that are to be excited by light will be relatively photostable and will not react efficiently with singlet oxygen. Several structural features are present in most useful sensitizers. Most sensitizers have at least one and frequently three or more conjugated double or triple bonds held in a rigid, frequently aromatic structure. They will frequently contain at least one group that accelerates intersystem crossing such as a carbonyl or imine group or a heavy atom selected from rows 3-6 of the periodic table, especially iodine or bromine, or they may have extended aromatic structures. Typical sensitizers include acetone, benzophenone, 9-thioxanthone, eosin, 9,10-dibromoanthracene, methylene blue, metallo-porphyrins, such as hematoporphyrin, phthalocyanines, chlorophylls, rose bengal, buckminsterfullerene, etc., and derivatives of these compounds having substituents of 1 to 50 atoms for rendering such compounds more lipophilic or more hydrophilic and/or as attaching groups for attachment. Examples of other photosensitizers that may be utilized are those that have the above properties and are enumerated in N. J. Turro, Molecular Photochemistry, page 132, W. A. Benjamin Inc., N.Y. 1965.

[1515] In some embodiments, the photosensitizers are relatively non-polar to assure dissolution into a lipophilic member when the photosensitizer is incorporated in an oil droplet, liposome, latex particle, etc.

[1516] In some embodiments, the photosensitizers suitable for use herein include other substances and compositions that can produce singlet oxygen with or without activation by an external light source. Thus, for example, molybdate (MoO.sub.4.sup.=) salts and chloroperoxidase and myeloperoxidase plus bromide or chloride ion (Kanofsky, J. Biol. Chem. (1983) 259 5596) have been shown to catalyze the conversion of hydrogen peroxide to singlet oxygen and water. Either of these compositions can, for example, be included in particles and used in the assay method wherein hydrogen peroxide is included as an ancillary reagebly, chloroperoxidase is bound to a surface and molybdate is incorporated in the aqueous phase of a liposome. Also included within the scope of the invention as photosensitizers are compounds that are not true sensitizers but which on excitation by heat, light, or chemical activation will release a molecule of singlet oxygen. The best known members of this class of compounds includes the endoperoxides such as 1,4-biscarboxyethyl-1,4-naphthalene endoperoxide, 9,10-diphenylanthracene-9,10-endoperoxide and 5,6,11,12-tetraphenyl naphthalene 5,12-endoperoxide. Heating or direct absorption of light by these compounds releases singlet oxygen.

[1517] A chemiluminescent compound as used herein refers to a substance that undergoes a chemical reaction with singlet oxygen to form a metastable intermediate that can decompose with the simultaneous or subsequent emission of light within the wavelength range of 250 to 1200 nm. Exemplary chemiluminescent compounds suitable for use include those described in U.S. Pat. Nos. 6,251,581 and 7,709,273, and Patent Cooperatio Treaty (PCT) International Application Publication No. WO1999/042838. Examplery chemiluminescent compound includes the following:

TABLE-US-00028 Emission Chemiluminescer Half-Life Max Thioxene + Diphenyl anthracence: 0.6 seconds 430 nm Thioxene + Umbelliferone derivative 0.6 seconds 500 nm Thioxene + Europium chelate 0.6 seconds 615 nm Thioxene + Samarium Chelate 0.6 seconds 648 nm Thioxene + terbium Chelate 0.6 seconds 540 nm N-Phenyl Oxazine + Umbelliferone 30 seconds 500 nm derivative N-Phenyl Oxazine + Europium chelate 30 seconds 613 nm N-phenyl Oxazine + Samarium Chelate 30 seconds 648 nm N-phenyl Oxazine + terbium Chelate 30 seconds 540 nm Dioxene + Umbelliferone derivative 300 seconds 500 nm Dioxene + Europium chelate 300 seconds 613 nm Dioxene + Samarium Chelate 300 seconds 648 nm N-phenyl Oxazine + terbium Chelate 300 seconds 540 nm

[1518] All of the above mentioned applications are hereby expressly incorporated by reference herein in their entireties. Emission will usually occur without the presence of an energy acceptor or catalyst to cause decomposition and light emission. In some embodiments, the intermediate decomposes spontaneously without heating or addition of ancillary reagents following its formation. However, addition of a reagent after formation of the intermediate or the use of elevated temperature to accelerate decomposition will be required for some chemiluminescent compounds. The chemiluminescent compounds are usually electron rich compounds that react with singlet oxygen, frequently with formation of dioxetanes or dioxetanones. Exemplary of such compounds are enol ethers, enamines, 9-alkylidenexanthans, 9-alkylidene-N-alkylacridans, aryl vinyl ethers, dioxenes, arylimidazoles and lucigenin. Other chemiluminescent compounds give intermediates upon reaction with singlet oxygen, which subsequently react with another reagent with light emission. Exemplary compounds are hydrazides such as luminol and oxalate esters.

[1519] The chemiluminescent compounds of interest will generally emit at wavelengths above 300 nanometers and usually above 400 nm. Compounds that alone or together with a fluorescent molecule emit light at wavelengths beyond the region where serum components absorb light will be of particular use. The fluorescence of serum drops off rapidly above 500 nm and becomes relatively unimportant above 550 nm. Therefore, when the analyte is in serum, chemiluminescent compounds that emit light above 550 nm, e.g., above 600 nm may be suitable for use. In order to avoid autosensitization of the chemiluminescent compound, in some embodiments, the chemiluminescent compounds do not absorb light used to excite the photosensitizer. In some embodiments, the sensitizer is excited with light wavelengths longer than 500 nm, it will therefore be desirable that light absorption by the chemiluminescent compound be very low above 500 nm.

[1520] Where long wave length emission from the chemiluminescent compound is desired, a long wavelength emitter such as a pyrene, bound to the chemiluminescent compound can be used. Alternatively, a fluorescent molecule can be included in the medium containing the chemiluminescent compound. In some embodiments, fluorescent molecules will be excited by the activated chemiluminescent compound and emit at a wavelength longer than the emission wavelength of the chemiluminescent compound, usually greater that 550 nm. It is usually also desirable that the fluorescent molecules do not absorb at the wavelengths of light used to activate the photosensitizer. Examples of useful dyes include rhodamine, ethidium, dansyl, Eu(fod).sub.3, Eu(TTA).sub.3, Ru(bpy).sub.3.sup.++ (wherein bpy=2,2-dipyridyl, etc. In general these dyes act as acceptors in energy transfer processes and in some embodiments, have high fluorescent quantum yields and do not react rapidly with singlet oxygen. They can be incorporated into particles simultaneously with the incorporation of the chemiluminescent compound into the particles.

[1521] In some embodiments, the disclosure provides diffractive optics detection technology that can be used with, for example, ingestible device technology. In certain embodiments, an ingestible device includes the diffractive optics technology (e.g., diffractive optics detection system). In certain embodiments, the disclosure provides diffractive optics technology (e.g., diffractive optics detection systems) that are used outside the body of subject. As an example, an ingestible device can be used to obtain one more samples in the body (e.g., in the gastrointestinal tract) of a subject, and the diffractive optics technology can be used to analyze the sample(s). Such analysis can be performed in vivo (e.g., when the ingestible device contains the diffractive optics).

[1522] Diffraction is a phenomenon that occurs due to the wave nature of light. When light hits an edge or passes through a small aperture, it is scattered in different directions. But light waves can interfere to add (constructively) and subtract (destructively) from each other, so that if light hits a non-random pattern of obstacles, the subsequent constructive and destructive interference will result in a clear and distinct diffraction pattern. A specific example is that of a diffraction grating, which is of uniformly spaced lines, typically prepared by ruling straight, parallel grooves on a surface. Light incident on such a surface produces a pattern of evenly spaced spots of high light intensity. This is called Bragg scattering, and the distance between spots (or Bragg scattering peaks) is a unique function of the diffraction pattern and the wavelength of the light source. Diffraction gratings, like focusing optics, can be operated in both transmission and reflection modes.

[1523] In general, the light used in the diffractive optics can be of any appropriate wavelength. Exemplary wavelengths include visible light, infrared red (IR) and ultraviolet (UV). Optionally, the light can be monochromatic or polychromatic. The light can be coherent or incoherent. The light can be collimated or non-collimated. In some embodiments, the light is coherent and collimated. Generally, any appropriate light source may be used, such as, for example, a laser (e.g., a laser diode) or a light emitting diode. In some embodiments, the light source is a laser diode operating at 670 nm wavelength, e.g., at 3 mWatts power. Optionally, an operating wavelength of a laser diode can be 780 nm, e.g., when larger grating periods are used. In certain embodiments, the light source is a laser, such as, for example, a HeNe laser, a Nd:YVO4 laser, or an argon-ion laser. In some embodiments, the light source is a low power, continuous waver laser.

[1524] The diffracted light can be detected using any appropriate light detector(s). Examples of light detectors include photodetectors, such as, for example, position sensitive photodiodes, photomultiplier tubes (PMTs), photodiodes (PDs), avalanche photodiodes (APDs), charged-coupled device (CCD) arrays, and CMOS detectors. In some embodiments, the diffracted light is detected via one or more individual photodiodes.

[1525] In general, the diffraction grating is made of a material that is transparent in the wavelength of the radiation used to illuminate the sensor. Any appropriate material may be used for the diffraction grating substrate, such as glass or a polymer. Exemplary polymers include polystyrene polymers (PSEs), cyclo-olefin polymers (COPs), polycarbonate polymers, polymethyl methacrylates, and methyl methacrylate styrene copolymers. Exemplary COPs include Zeonex (e.g., Zeonex E48R, Zeonex F52R).

[1526] The light may be incident on the diffraction grating any appropriate angle. In some embodiments, the light is incident on the diffraction grating with an angle of incidence of from 30 to 80 (e.g., from 40 to 80, from 50 to 70, from 55 to 65, 60). Optionally, the system is configured so that that diffractive grating and light source can move relative to each other

[1527] In general, the light detector can be positioned with respect to the diffractive grating so that the diffraction grating can be illuminated at a desired angle of incidence and/or so that diffracted light can be detected at a desired angle and/or so that diffracted light of a desired order can be detected.

[1528] The period P of the diffraction grating can be selected as desired. In some embodiments, the period P is from 0.5 microns to 50 microns (e.g., from one micron to 15 microns, from one micron to five microns). In some embodiments, the grating is a repeating patter of 1.5 micron and 4.5 micron lines with a period of 15 microns.

[1529] The height h of the diffraction grating can be selected as desired. In certain embodiments, the height h is from one nanometer to about 1000 nanometers (e.g., from about five nanometers to about 250 nanometers, from five nanometers to 100 nanometers).

[1530] In general, the diffractive optics can be prepared using any appropriate method, such as, for example, surface ablation, photolithograph (e.g., UV photolithography), laser etching, electron beam etching, nano-imprint molding, or microcontact printing.

[1531] Optionally, the diffractive optics system can include one or more additional optical elements, such as, for example, one or more mirrors, filters and/or lenses. Such optical elements can, for example, be arranged between the light source and the diffractive grating and/or between the diffractive grating and the detector.

[1532] In some of the embodiments of the devices described herein, a primary binding partner specifically binds to a secondary binding partner through non-covalent interactions (e.g., electrostatic, van der Waals, hydrophobic effect). In some embodiments, a primary binding partner specifically binds to a secondary binding partner via a covalent bond (e.g., a polar covalent bond or a non-polar covalent bond). In some embodiments of any of the devices described herein, the primary and the secondary binding partner can be interchanged. For example, the primary binding partner can be biotin, or a derivative thereof, and the secondary binding partner is avidin, or a derivative thereof. In other examples, the primary binding partner can be avidin, or a derivative thereof, and the secondary binding partner is biotin.

[1533] In some embodiments, the binding of the primary and the secondary binding partner is essentially irreversible. In some embodiments, the binding of the primary and the secondary binding partner is reversible. In some embodiments, the primary binding partner is CaptAvidin biotin-binding protein and the secondary binding partner is biotin, or vice versa. In some embodiments, the primary binding partner is DSB-X biotin and the secondary binding partner is avidin, or vice versa. In some embodiments, the primary binding partner is desthiobiotin and the secondary binding partner is avidin, or vice versa (Hirsch et al., Anal Biochem. 308(2):343-357, 2002). In some embodiments, the primary binding partner is glutathione (GSH) or a derivative thereof, and the secondary binding partner is glutathione-S-transferase (GST).

[1534] In some embodiments, the primary binding partner can bind to a target analyte that is a nucleic acid (e.g., a DNA molecule, a RNA molecule). In some embodiments, the primary binding partner comprises a portion of a nucleic acid that is complementary to the nucleic acid sequence of the target analyte.

[1535] In some embodiments of any of the devices described herein, the device can include a label that binds to the target analyte and does not prevent binding of the target analyte to the primary binding partner. In some embodiments, the label can amplify the diffraction signal of the target analyte.

[1536] In some embodiments, the label is from about 1 nm to 200 nm (e.g., about 50 nm to about 200 nm).

[1537] In some embodiments, the label (e.g., any of the labels described herein) includes one or more antibodies (e.g., any of the antibodies and/or antibody fragments described herein).

[1538] In some embodiments, the label is a nanoparticle (e.g., a gold nanoparticle) that includes the primary binding partner that has a nucleic acid sequence that is complementary to the target analyte, and is covalently linked to the nanoparticle.

[1539] One or more additional steps can be performed in any of the methods described herein. In some embodiments, the one or more additional steps are performed: prior to the binding of the primary binding partner to the secondary binding partner, after the binding of the primary binding partner to the secondary binding partner, prior to the binding of the primary binding partner to the target analyte, or after the binding of the primary binding partner to the target analyte.

[1540] In some embodiments of any of the methods described herein, the determining step (during which the primary binding partner binds to the target analyte is detected) can occur in at least 15 seconds. In some embodiments, the binding of the primary binding partner to the target analyte can occur during a period of time of, for example, five at least seconds.

[1541] In some embodiments, the one or more additional steps can include: a blocking of the sensors step, at least one wash step, a capturing step, and/or a filtering step. In some embodiments, the blocking step can include blocking a sensor within the ingestible device with a solution comprising at least 1% bovine serum albumin (BSA) in a buffered solution (e.g., phosphate buffered saline (PBS), Tris buffered saline (TBS)). In some embodiments, the at least one wash step can include washing with a buffered solution (e.g., phosphate buffered saline (PBS), Tris buffered saline (TBS)). In general, blocking is performed during capsule manufacture, rather than in vivo.

[1542] In some embodiments, the capturing step includes enriching the target analyte. In some embodiments, the capturing step includes physically separating the target analyte from the remaining sample using a filter, a pore, or a magnetic bead. In some embodiments, the target analyte is captured by size exclusion.

[1543] In some embodiments, the disclosure provides methods of obtaining, culturing, and/or detecting target cells and/or target analytes in vivo within the gastrointestinal (GI) tract or reproductive tract of a subject. Associated devices are also disclosed. The methods and devices described provide a number of advantages for obtaining and/or analyzing fluid samples from a subject. In some embodiments, diluting the fluid sample increases the dynamic range of analyte detection and/or reduces background signals or interference within the sample. For example, interference may be caused by the presence of non-target analytes or non-specific binding of a dye or label within the sample. In some embodiments, culturing the sample increases the concentration of target cells and/or target analytes produced by the target cells thereby facilitating their detection and/or characterization.

[1544] In certain embodiments, the methods and devices a described herein may be used to obtain information regarding bacteria populations in the GI tract of a subject. This has a number of advantages and is less invasive than surgical procedures such as intubation or endoscopy to obtain fluid samples from the GI tract. The use of an ingestible device as described herein also allows for fluid samples to be obtained and data to be generated on bacterial populations from specific regions of the GI tract.

[1545] In some embodiments, the methods and devices described herein may be used to generate data such as by analyzing the fluid sample, dilutions thereof or cultured samples for one or more target cells and/or target analytes. The data may include, but is not limited to, the types of bacteria present in the fluid sample or the concentration of bacteria in specific regions of the GI tract. Such data may be used to determine whether a subject has an infection, such as Small Intestinal Bacterial Overgrowth (SIBO), or to characterize bacterial populations within the GI tract for diagnostic or other purposes. Thus, in some embodiments, analytes disclosed herein are indicative of disorders of the gastrointestinal tract associated with anomalous bacterial populations.

[1546] For example, in one aspect, the data may include, but is not limited to, the concentration of bacteria in a specific region of the GI tract that is one or more of the duodenum, jejunum, ileum, ascending colon, transverse colon or descending colon. In one aspect, the specific region of the GI tract is the duodenum. In one aspect, the specific region of the GI tract is the jejunum. In one aspect, the specific region of the GI tract is the ileum. In one aspect, the specific region of the GI tract is the ascending colon. In one aspect, the specific region of the GI tract is the transverse colon. In one aspect, the specific region of the GI tract is the descending colon. In a related embodiment, the data may be generated every one or more days to monitor disease flare-ups, or response to the therapeutic agents disclosed herein.

[1547] Data may be generated after the device has exited the subject, or the data may be generated in vivo and stored on the device and recovered ex vivo. Alternatively, the data can be transmitted wirelessly from the device while the device is passing through the GI tract of the subject or in place within the reproductive tract of the subject.

[1548] In some embodiments, a method comprises: providing a device comprising one or more dilution chambers and dilution fluid; transferring all or part of a fluid sample obtained from the GI tract or reproductive tract of the subject into the one or more dilution chambers in vivo; and combining the fluid sample and the dilution fluid to produce one or more diluted samples in the one or more dilution chambers.

[1549] In certain embodiments, a method comprises: providing an ingestible device comprising one or more dilution chambers; transferring all or part of a fluid sample obtained from the GI tract into the one or more dilution chambers comprising sterile media; culturing the sample in vivo within the one or more dilution chambers to produce one or more cultured samples; and detecting bacteria in the one or more cultured samples.

[1550] In some embodiments, a method comprises: providing a device comprising one or more dilution chambers; transferring all or part of a fluid sample obtained from the GI tract or reproductive tract into the one or more dilution chambers; combining all or part of the fluid sample with a dilution fluid in the one or more dilution chambers; and detecting the target analyte in the one or more diluted samples.

[1551] In certain embodiments, a device comprises: one or more dilution chambers for diluting a fluid sample obtained from the GI tract or reproductive tract; and dilution fluid for diluting the sample within the one or more dilution chambers.

[1552] In some embodiments, the device comprises: one or more dilution chambers for culturing a fluid sample obtained from the GI tract; sterile media for culturing the sample within the one or more dilution chambers; and a detection system for detecting bacteria.

[1553] In certain embodiments, a device comprises: one or more dilution chambers for culturing a fluid sample obtained from the GI tract; sterile media for culturing the sample within the one or more dilution chambers; and a detection system for detecting bacteria.

[1554] Also provided is the use of a device as described herein for diluting one or more samples obtained from the GI tract or reproductive tract of a subject. In one embodiment, there is provided the use of an ingestible device as described herein for detecting target cells and/or target analytes in vivo within the gastrointestinal (GI) tract of a subject.

[1555] Further provided is a system comprising a device as described herein and a base station. In one embodiment, the device transmits data to the base station, such as data indicative of the concentration and/or types of bacteria in the GI tract of the subject. In one embodiment, the device receives operating parameters from the base station. Some embodiments described herein provide an ingestible device for obtaining one or more samples from the GI tract or reproductive tract of a subject and diluting and/or culturing all or part of the one or more samples. The ingestible device includes a cylindrical rotatable element having a port on the wall of the cylindrical rotatable element. The ingestible device further includes a shell element wrapping around the cylindrical rotatable element to form a first dilution chamber between the cylindrical rotatable element and the shell element. The shell element has an aperture that exposes a portion of the wall of the cylindrical rotatable element to an exterior of the ingestible device.

[1556] In certain embodiments, the medical device comprises one or more dilution chambers for receiving a fluid sample from the GI tract or reproductive tract of a subject or a dilution thereof. In some embodiments, one or more dilutions of the fluid sample are cultured in one or more dilution chambers. In certain embodiments, the dilution chambers each define a known volume, optionally the same volume or different volumes. In some embodiments, the dilution chambers define a fluid volume ranging from about 10 L to about 1 mL. The dilution chambers may define a fluid volume less than or equal to about 500 L, less than or equal to about 250 L, less than or equal to about 100 L, or less than or equal to about 50 L. In certain embodiments, the dilution chambers define a fluid volume of greater than or equal to about 10 L, greater than or equal to about 20 L, greater than or equal to about 30 L, or greater than or equal to about 50 L. In some embodiments, the dilution chambers define a fluid volume between about 10 L and 500 L, between about 20 L and 250 L, between about 30 L and 100 L or about 50 L.

[1557] In some embodiments, dilution fluid in the device is combined with all or part of the fluid sample, or dilution thereof, to produce one or more dilutions. In certain embodiments, the dilution fluid is sterile media suitable for culturing one or more target cells within the dilution chambers.

[1558] In certain embodiments, the one or more dilution chambers may be filled with the dilution fluid prior to a patient ingesting the ingestible device. In some embodiments, the dilution fluid may be added into the one or more dilution chambers in vivo from a reservoir of the ingestible device. Sampling and dilution of the GI fluid sample may take place in vivo. For example, an actuator of the ingestible device may pump the dilution fluid from the reservoir into a dilution chamber when it is determined that the ingestible device is located at a predetermined location within the GI tract. In some embodiments, the dilution chambers each contain a volume of sterile media suitable for culturing a fluid sample from the GI tract or reproductive tract. In certain embodiments, the dilution chambers are at least 95%, at least 97%, at least 98%, or at least 99% full of sterile media. In some embodiments, the dilution chambers each contain oxygen to facilitate aerobic bacteria growth. In certain embodiments, a non-dilution chamber comprises oxygen and is added to one or more of the dilution chambers to facilitate aerobic bacteria growth.

[1559] In some embodiments, the culturing may take place in vivo immediately after the GI fluid sample has been diluted. Or alternatively, the culturing may take place ex vivo, e.g., when the ingestible device has been evacuated and recovered such that the dilution chamber containing the diluted GI fluid sample may be extracted and the culturing may be performed in a laboratory. The recovery of the ingestible device may be performed in a similar manner as embodiments described in U.S. Provisional Application No. 62/434,188, filed on Dec. 14, 2016, which is herein expressly incorporated by reference in its entirety.

[1560] As used herein culturing refers to maintaining target cells in an environment that allows a population of one or more target cells to increase in number through cell division. For example, in some embodiments, culturing may include combining the cells with media in an dilution chamber at a temperature that permits cell growth, optionally a temperature found in vivo within the GI tract or reproductive tract of a subject. In certain embodiments, the cells are cultured at a temperature between about 35 C. and 42 C.

[1561] As used herein dilution fluid refers to a fluid within the device for diluting a fluid sample from the GI tract or reproductive tract. In some embodiments, the dilution fluid is an aqueous solution. In certain embodiments, the dilution fluid comprises one or more agents that promote or inhibit the growth of an organism, such as a fungus or bacteria. In some embodiments, the dilution fluid comprises one or more agents that facilitate the detection of a target analyte, such as dyes or binding agents for target analytes.

[1562] In some embodiments, the dilution fluid is a sterile media. As used herein, sterile media refers to media that does not contain any viable bacteria or other cells that would grow and increase in number through cell division. Media may be rendered sterile by various techniques known in the art such as, but not limited to, autoclaving and/or preparing the media using asceptic techniques. In certain embodiments, the media is a liquid media. Examples of media suitable for culturing bacteria include nutrient broth, Lysogeny Broth (LB) (also known as Luria Broth), Wilkins chalgren, and Tryptic Soy Broth (TSB), Other growth or culture media known in the art may also be used in the methods and devices described herein. In some embodiments, the media has a carbon source, such as glucose or glycerol, a nitrogen source such as ammonium salts or nitrates or amino acids, as well as salts and/or trace elements and vitamins required for microbial growth. In certain embodiments, the media is suitable for maintaining eukaryotic cells. In some embodiments, the media comprises one or more agents that promote or inhibit the growth of bacteria, optionally agents that promote or inhibit the growth of specific types of bacteria.

[1563] In certain embodiments, the media is a selective media. As used herein, selective media refers to a media that allows certain types of target cells to grow and inhibits the growth of other organisms. Accordingly, the growth of cells in a selective media indicates the presence of certain types of cells within the cultured sample. For example, in some embodiments, the media is selective for gram-positive or gram-negative bacteria. In certain embodiments, the media contains crystal violet and bile salts (such as found in MacConkey agar) that inhibit the growth of gram-positive organisms and allows for the selection and isolation of gram-negative bacteria. In some embodiments, the media contains a high concentration of salt (NaCl) (such as found in Mannitol salt agar) and is selective for Gram-positive bacteria. In some embodiments, the media selectively kills eukaryotic cells or only grows prokaryotic cells, for example, using a media comprising Triton X-100. In certain embodiments, the media selectively kills prokaryotic cells (or alternatively only grows eukaryotic cells), for example, using a media that comprises antibiotics.

[1564] In some embodiments, the media is an indicator media. As used herein, indicator media refers to a media that contains specific nutrients or indicators (such as, but not limited to neutral red, phenol red, eosin , or methylene blue) that produce a detectable signal when a certain type of cells are cultured in the indicator media.

[1565] In some embodiments, the disclosure provides a composition comprising a dye and optionally a reagent for selective lysis of eukaryotic cells. In certain embodiments, the composition comprises both a dye and a reagent for selective lysis of eukaryotic cells. In some embodiments, the composition further comprises one or more reagents independently selected from the group consisting of: a second reagent for selective lysis of eukaryotic cells (e.g., Triton X-100), an electrolyte (e.g., MgCl.sub.2), an anti-fungi reagent (e.g., amphotericin-B), and an antibiotic. In some embodiments, the composition comprises water and is in the form of an aqueous solution. In some embodiments, the composition is a solid or semi-solid. In some embodiments, the compositions described here are suitable for use in a kit or device for detecting or quantifying viable bacterial cells in a sample. In some embodiments, such a device is an ingestible device for detecting or quantifying viable bacterial cells in vivo (e.g., in the GI tract). In some embodiments, viable bacterial cells in a sample are detected or quantified in the presence of one or more antibiotics to determine antibiotic resistance of the bacteria in the sample. In some embodiments, anomalous bacterial populations in a sample may be detected or quantified, for example through the use of one a composition comprising a dye as disclosed herein, to determine whether a subject has an infection, such as Small Intestinal Bacterial Overgrowth (SIBO), or to characterize bacterial populations within the GI tract for diagnostic or other purposes.

[1566] In some embodiments, a method comprises: (a) contacting the sample with a composition as described herein; and (b) measuring total fluorescence or rate of change of fluorescence as a function of time of said sample, thereby detecting viable bacterial cells in said sample. In some embodiments, a control as described herein may be employed in the method. In some embodiments, the total fluorescence or the rate of change of fluorescence as a function of time of the sample is measured over multiple time points for an extended period of time in step (b), thereby detecting viable bacterial cells in said sample. In some embodiments, the method further comprises correlating the total fluorescence or the rate of change of fluorescence as a function of time determined in step (b) to the number of viable bacterial cells in the sample. In some embodiments, the rate of change of fluorescence as a function of time of the sample measured over multiple time points is determined and compared to the rate of change of fluorescence as a function of time of a control measured over the same time points to determine the number of viable bacterial cells in the sample. In some embodiments, the method does not require ex vivo plating or culturing. In some embodiments, the method does not require aspiration. In some embodiments, the method is performed in vivo (e.g., in an ingestible device in vivo). In some embodiments, the method comprises communicating the results of the onboard assay(s) to an ex vivo receiver.

[1567] In certain embodiments, a kit comprises a composition as described herein and instructions, e.g., for detecting or quantifying viable bacterial cells in a sample. In some embodiments, a device comprises a composition as described herein, e.g., for detecting or quantifying viable bacterial cells in a sample. The detection of live cells, as opposed to the detection of bacterial components (such as endotoxins) which can be present in the sample environment and lead to conflicting results, is the gold standard of viable plate counting and represents one of the advantages of the compositions and methods described herein.

[1568] The systems employ methods, compositions and detection systems found to accurately and reliably correlate fluorescence to total bacteria count (TBC) in an autonomous, ingestible device, or other similarly-sized device. The compositions include novel combinations of dyes, buffers and detergents that allow for the selective staining of viable bacterial cells in samples that comprise non-bacterial cells and other components that otherwise make detecting or quantifying live bacterial cells challenging. In some embodiments, the systems allow for bacteria to be quantified in near real-time and the results to be shared telemetrically outside of the device.

[1569] In certain embodiments, the disclosure provides a method of assessing or monitoring the need to treat a subject suffering from or at risk of overgrowth of bacterial cells in the gastrointestinal tract, which comprises: (a) obtaining a sample from the gastrointestinal tract of said subject; (b) contacting the sample with a composition as described herein; (c) measuring total fluorescence or rate of change of fluorescence as a function of time of said sample; and (d) correlating the total fluorescence or the rate of change of fluorescence as a function of time measured in step (c) to the number of viable bacterial cells in the sample, wherein the number of the viable bacterial cells determined in step (e) greater than about 105 CFU/mL indicates a need for treatment, e.g., with an antibiotic agent as described herein. In some embodiments, a control as described herein may be employed in the method. In some embodiments, the total fluorescence or the rate of change of fluorescence as a function of time of the sample is measured over multiple time points for an extended period of time in step (c). In some embodiments, the rate of change of fluorescence as a function of time of the sample measured over multiple time points is determined and compared to the rate of change of fluorescence as a function of time of a control measured over the same time points to determine the number of viable bacterial cells in the sample. In some embodiments, the method does not require ex vivo plating or culturing. In some embodiments, the method does not require aspiration. In some embodiments, the method is performed in vivo (e.g., in an ingestible device in vivo). In some embodiments, the method comprises communicating the results of the onboard assay(s) to an ex vivo receiver. In some embodiments, the method may be further used to monitor the subject after the treatment (e.g., with an antibiotic). In some embodiments, the method may be used to assess the efficacy of the treatment. For example, efficacious treatment may be indicated by the decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. Efficacy of the treatment may be evaluated by the rate of decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. In some embodiments, the method may be used to detect infection with antibiotic-resistant strains of bacteria in a subject. For instance, such infection may be indicated where the number of viable bacterial cells in a sample from the GI tract of the subject does not substantially decrease after antibiotic treatment.

[1570] In some embodiments, the disclosure provides an absorbable material, (e.g., absorbable sponge), having absorbed therein a composition as described herein. In some embodiments, the absorbable sponge is Ahlstrom Grade 6613H (Lot 150191) or Porex PSU-567, having absorbed therein a composition as described herein. In some embodiments, the absorbable sponge may be prepared by injecting into the absorbable sponge an aqueous solution comprising a composition as described herein, and optionally further comprising a step of drying the resulting absorbable sponge.

[1571] In certain embodiments, the disclosure provides a method for detecting the presence of viable bacterial cells in a sample, which comprises: (a) fully or partially saturating an absorbable sponge as described herein, or an absorbable sponge prepared as described herein, with the sample; and (b) measuring total fluorescence or rate of change of fluorescence as a function of time of the fully or partially saturated sponge prepared in step (a), thereby detecting viable bacterial cells. In some embodiments, a control as described herein may be employed in the method. In some embodiments, the total fluorescence or the rate of change of fluorescence as a function of time of the fully or partially saturated sponge is measured over multiple time points for an extended period of time in step (b), thereby detecting viable bacterial cells in said sample. In some embodiments, the method further comprises correlating the total fluorescence or the rate of change of fluorescence as a function of time measured in step (b) to the number of viable bacterial cells in the sample. In some embodiments, the rate of change of fluorescence as a function of time of the fully or partially saturated sponge measured over multiple time points is determined and compared to the rate of change of fluorescence as a function of time of a control measured over the same time points to determine the number of viable bacterial cells in the sample. In some embodiments, the method does not require ex vivo plating or culturing. In some embodiments, the method does not require aspiration. In some embodiments, the method is performed in vivo (e.g., in an ingestible device in vivo). In some embodiments, the method comprises communicating the results of the onboard assay(s) to an ex vivo receiver.

In one aspect, provided herein is a kit comprising an absorbable sponge as described herein and instructions, e.g., for detecting or quantifying viable bacterial cells in a sample. In another aspect, provided herein is a device comprising an absorbable sponge as described herein, e.g., for detecting or quantifying viable bacterial cells in a sample.

[1572] In certain embodiments, the disclosure provides a method of assessing or monitoring the need to treat a subject suffering from or at risk of overgrowth of bacterial cells in the gastrointestinal tract, which comprises: (a) obtaining a sample from the gastrointestinal tract of said subject; (b) fully or partially saturating an absorbable sponge described herein, or an absorbable sponge prepared as described herein, with the sample; (c) measuring total fluorescence or rate of change of fluorescence as a function of time of the fully or partially saturated sponge prepared in step (b); (d) correlating the total fluorescence or the rate of change of fluorescence as a function of time measured in step (c) to the number of viable bacterial cells in the sample, wherein the number of the viable bacterial cells as determined in step (e) greater than about 10.sup.5 CFU/mL indicates a need for treatment, e.g., with an antibiotic agent as described herein. In some embodiments, a control as described herein may be employed in the method. In some embodiments, the total fluorescence or the rate of change of fluorescence as a function of time of the fully or partially saturated sponge is measured over multiple time points for an extended period of time in step (c). In some embodiments, the rate of change of fluorescence as a function of time of the fully or partially saturated sponge measured over multiple time points is determined and compared to the rate of change of fluorescence as a function of time of a control measured over the same time points to determine the number of viable bacterial cells in the sample. In some embodiments, the method does not require ex vivo plating or culturing. In some embodiments, the method does not require aspiration. In some embodiments, the method is performed in vivo (e.g., in an ingestible device in vivo). In some embodiments, the method comprises communicating the results of the onboard assay(s) to an ex vivo receiver. In some embodiments, the method may be further used to monitor the subject after the treatment (e.g., with an antibiotic). In some embodiments, the method may be used to assess the efficacy of the treatment. For example, efficacious treatment may be indicated by the decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. Efficacy of the treatment may be evaluated by the rate of decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. In some embodiments, the method may be used to detect infection with antibiotic-resistant strains of bacteria in a subject. For instance, such infection may be indicated where the number of viable bacterial cells in a sample from the GI tract of the subject does not substantially decrease after antibiotic treatment

[1573] In certain embodiments, the disclosure provides and ingestible device comprising a housing; a first opening in the wall of the housing; a second opening in the first end of the housing; and a chamber connecting the first opening and the second opening, wherein at least a portion of the chamber forms a sampling chamber within the ingestible device. In some embodiments, the sampling chamber is configured to hold an absorbable sponge described herein. In some embodiments, the sampling chamber is configured to hold a sample obtained from a gastrointestinal (GI) tract of a body. In some embodiments, the ingestible device is individually calibrated (for example, by comparing to a positive or negative control as described herein), wherein the fluorescent properties of the absorbable sponge held in the sampling chamber of the device are determined prior to the introduction of the sample. The ingestible device as described herein is useful for detecting or quantifying viable bacterial cells in vivo. In some embodiments, provided herein is a method for detecting or quantifying viable bacterial cells in a GI tract sample in vivo using an ingestible device as described herein. In some embodiments, provided herein is a method of assessing or monitoring the need to treat a subject suffering from or at risk of overgrowth of bacterial cells in the GI tract in vivo using an ingestible device as described herein. In some embodiments, provided herein is a method of altering the treatment regimen of a subject suffering from or at risk of overgrowth of bacterial cells in the GI tract in vivo using an ingestible device as described herein. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the duodenum. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the jejunum. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the ileum. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the ascending colon. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the transverse colon. In one aspect, the subject is a subject suffering from or at risk of overgrowth of bacterial cells in the descending colon. In some embodiments, the method may be further used to monitor the subject after the treatment (e.g., with an antibiotic). In some embodiments, the method may be used to assess the efficacy of the treatment. For example, efficacious treatment may be indicated by the decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. Efficacy of the treatment may be evaluated by the rate of decrease of the number of viable bacterial cells in a sample from the GI tract of the subject post-treatment. In some embodiments, the method may be used to detect infection with antibiotic-resistant strains of bacteria in a subject. For instance, such infection may be indicated where the number of viable bacterial cells in a sample from the GI tract of the subject does not substantially decrease after antibiotic treatment. In some embodiments, the method is performed autonomously and does not require instructions, triggers or other inputs from outside the body after the device has been ingested.

[1574] Eukaryotic as recited herein relates to any type of eukaryotic organism excluding fungi, such as animals, in particular animals containing blood, and comprises invertebrate animals such as crustaceans and vertebrates. Vertebrates comprise both cold-blooded (fish, reptiles, amphibians) and warm blooded animal (birds and mammals). Mammals comprise in particular primates and more particularly humans

[1575] Selective lysis as used herein is obtained in a sample when the percentage of bacterial cells in that sample that remain intact is significantly higher (e.g. 2, 5, 10, 20, 50, 100, 250, 500, or 1,000 times more) than the percentage of the eukaryotic cells in that sample that remain intact, upon treatment of or contact with a composition or device as described herein.

[1576] In some embodiments, the dye suitable for use herein is a dye that is capable of being internalized by a viable cell, binding to or reacting with a target component of the viable cell, and having fluorescence properties that are measurably altered when the dye is bound to or reacted with the target component of the viable cell. In some embodiments, the dye herein is actively internalized by penetrating viable cells through a process other than passible diffusion across cell membranes. Such internalization includes, but is not limited to, internalization through cell receptors on cell surfaces or through channels in cell membranes. In some embodiments, the target component of a viable cell to which the dye is bound to or reacted with is selected from the group consisting of: nucleic acids, actin, tubulin, enzymes, nucleotide-binding proteins, ion-transport proteins, mitochondria, cytoplasmic components, and membrane components. In some embodiments, the dye suitable for use herein is a fluorogenic dye that is capable of being internalized and metabolized by a viable cell, and wherein said dye fluoresces when metabolized by the viable cell. In some embodiments, the dye is a chemiluminescent dye that is capable of being internalized and metabolized by a viable cell, and wherein said dye becomes chemiluminescent when metabolized by the viable cell.

[1577] In some embodiments, the composition comprises a dye that fluoresces when bond to nucleic acids. Examples of such dyes include, but are not limited to, acridine orange (U.S. Pat. No. 4,190,328); calcein-AM (U.S. Pat. No. 5,314,805); DAPI; Hoechst 33342; Hoechst 33258; PicoGreen; SYTO 16; SYBR Green I; Texas Red; Redmond Red; Bodipy Dyes; Oregon Green; ethidium bromide; and propidium iodide.

[1578] In some embodiments, the composition comprises a lipophilic dye that fluoresces when metabolized by a cell. In some embodiments, the dye fluoresces when reduced by a cell or a cell component. Examples of dyes that fluoresce when reduced include, but are not limited to, resazurin; C.sup.12-resazurin; 7-hydroxy-9H-(1,3 dichloro-9,9-dimethylacridin-2-ol)N-oxide; 6-chloro-9-nitro-5-oxo-5H-benzo[a]phenoxazine; and tetrazolium salts. In some embodiment, the dye fluoresces when oxidized by a cell or a cell component. Examples of such dyes include, but are not limited to, dihydrocalcein AM; dihydrorhodamine 123; dihydroethidium; 2,3,4,5,6-pentafluorotetramethyldihydrorosamine; and 3-(p-aminophenyl) fluorescein.

[1579] In some embodiments, the composition comprises a dye that becomes chemiluminescent when oxidized by a cell or a cell component, such as luminol.

[1580] In some embodiments, the composition comprises a dye that fluoresces when de-acetylated and/or oxidized by a cell or a cell component. Examples of such dyes include, but are not limited to, dihydrorhodamines; dihydrofluoresceins; 2,7-dichlorodihydrofluorescein diacetate; 5-(and 6-)carboxy-2,7-dichlorodihydrofluorescein diacetate; and chloromethyl-2,7-dichlorodihydrofluorescein diacetate acetyl ester.

[1581] In some embodiments, the composition comprises a dye that fluoresces when reacted with a peptidase. Examples of such dyes include, but are not limited to, (CBZ-Ala-Ala-Ala-Ala)2-R110 elastase 2; (CBZ-Ala-Ala-Asp)2-R110 granzyme B; and 7-amino-4-methylcoumarin, N-CBZ-L-aspartyl-L-glutamyl-L-valyl-L-aspartic acid amide.

[1582] In some embodiments, the composition comprises a dye selected from the group consisting of resazurin, FDA, Calcein AM, and SYTO 9. In some embodiments, the dye is FDA or SYTO 9.

[1583] SYTO 9, when used alone, labels the nucleic acid of bacteria cells. The excitation/emission wavelengths for SYTO 9 is 480/500 nm, with the background remaining non-fluorescent. See, e.g., J. Appl. Bacteriol. 72, 410 (1992); Lett. Appl. Microbiol. 13, 58 (1991); Curr. Microbiol. 4, 321 (1980); J. Microbiol. Methods 13, 87 (1991); and Microbiol. Rev. 51, 365 (1987); and J. Med. Microbiol. 39, 147 (1993).

[1584] FDA is a non-polar, non-fluorescent compound that can cross the membranes of mammalian and bacterial cells. The acetyl esterases (present only within viable cells) hydrolyze the FDA into the fluorescent compound fluorescein. Fluorescein is a fluorescent polar compound that is retained within these cells. Living cells can be visualized in a photospectrometer when assayed with an excitation wavelength of 494 nm and an emission wavelength of 518 nm. See, e.g., Brunius, G. (1980). Technical aspects of the use of 3,6-Diacetyl fluorescein for vital fluorescent staining of bacteria. Current Microbiol. 4: 321-323; Jones, K. H. and Senft, J. A. (1985). An improved method to determine cellviability by simultaneous staining with fluorescein diacetate-propidium iodide. J. Histochem. Cytochem. 33: 77-79; Ross, R. D. , Joneckis, C. C., Ordonez, J. V, Sisk, A. M., Wu, R. K., Hamburger, A. W., and Nora, R. E. (1989). Estimation of cell survival by flow cytometric quantifcation of fluorescein diacetate/propidium iodide viable cell number. Cancer Research. 49: 3776-3782.

[1585] Calcein-AM, which is an acetoxylmethyl ester of calcein, is highly lipophilic and cell permeable. Calcein-AM in itself is not fluorescent, but the calcein generated by esterase in a viable cell emits a green fluorescence with an excitation wavelength of 490 nm and an emission of 515 nm. Therefore, Calcein-AM can only stain viable cells. See, e.g., Kimura, K., et al., Neurosci. Lett., 208, 53 (1998); Shimokawa, I., et al., J. Geronto., 51a, b49 (1998); Yoshida, S., et al., Clin. Nephrol., 49, 273 (1998); and Tominaga, H., et al., Anal. Commun., 36, 47 (1999).

[1586] Resazuirn (also known as Alamar Blue) is a blue compound that can be reduced to pink resorufin which is fluorescent. This dye is mainly used in viability assays for mammalian cells. C.sup.12-resazurin has better cell permeability than resazurin. When lipohilic C.sup.12-resazurin crosses the cell membranes, it is subsequently reduced by living cells to make a red fluorescent resorufin. The adsorption/emission of C.sup.12-resazurin is 563/587 nm. See, e.g., Appl Environ Microbiol 56, 3785 (1990); J Dairy Res 57, 239 (1990); J Neurosci Methods 70, 195 (1996); J Immunol Methods 210, 25 (1997); J Immunol Methods 213, 157 (1998); Antimicrob Agents Chemother 41, 1004 (1997).

[1587] In some embodiments, the composition optionally further comprises a reagent for selective lysis of eukaryotic cells. In some embodiments, the composition comprises a dye as described herein and a reagent for selective lysis of eukaryotic cells. In some embodiments, the reagent for selective lysis of eukaryotic cells is a detergent, such as a non-ionic or an ionic detergent. Examples of the reagent for selective lysis of eukaryotic cells include, but are not limited to, alkylglycosides, Brij 35 (C12E23 Polyoxyethyleneglycol dodecyl ether), Brij 58 (C16E20 Polyoxyethyleneglycol dodecyl ether), Genapol, glucanids such as MEGA-8, -9, -10, octylglucoside, Pluronic F127, Triton X-100 (C.sub.14H.sub.22O(C.sub.2H.sub.4O)n), Triton X-114 (C.sub.24H.sub.42O.sub.6), Tween 20 (Polysorbate 20) and Tween 80 (Polysorbate 80), Nonidet P40, deoxycholate, reduced Triton X-100 and/or Igepal CA 630. In some embodiments, the composition comprises a dye as described herein and deoxycholate (e.g., sodium deoxycholate) as a reagent for selective lysis of eukaryotic cells. In some embodiments, the composition comprises deoxycholate at a concentration selected from 0.0001% to 1 wt %. In some embodiments, the composition comprises deoxycholate at a concentration of 0.005 wt %. In some embodiments, the composition may comprise more than one reagent for selective lysis of eukaryotic cells.

[1588] In some embodiments, the composition may comprise two different reagents for selective lysis of eukaryotic cells. In some instances, when more than one selective lysis reagents are used, more effective and/or complete selective lysis of eukaryotic cells in a sample may be achieved. For example, the composition may comprise deoxycholate (e.g., sodium deoxycholate) and Triton X-100 as two different reagents for selective lysis of eukaryotic cells. In some embodiments, the composition comprises deoxycholate (e.g., sodium deoxycholate) at a concentration selected from 0.0001% to 1 wt % (e.g., 0.005 wt %) and Triton X-100 at a concentration selected from 0.1 to 0.05 wt %.

[1589] In some embodiments, after a sample (e.g., a biological sample) is treated or contacted with a composition comprising a dye and one or more reagents for selective lysis of eukaryotic cells as described herein, the eukaryotic cells (e.g., animal cells) in the sample are selectively lysed whereby a substantial percentage (e.g., more than 20%, 40%, 60%, 80%, 90% or even more that 95%) of the bacterial cells in the same sample remains intact or alive.

[1590] In some embodiments, the composition does not comprise a reagent for selective lysis of eukaryotic cells, and such a composition is useful for detecting or quantifying viable bacterial cells in a sample (e.g., an environmental sample such as a water sample) that does not contain any eukaryotic cells.

[1591] In some embodiments, the composition further comprises an electrolyte, such as a divalent electrolyte (e.g., MgCl.sub.2). In some embodiments, the composition comprises MgCl.sub.2 at a concentration selected from 0.1 mM to 100 mM (e.g., a concentration selected from 0.5 mM to 50 mM).

[1592] In some embodiments, the composition further comprises water and is in a form of an aqueous solution. In some embodiments, the composition has a pH selected from 5-8 (e.g., a pH selected from 6-7.8, such as pH being 6.0). In some embodiments, the composition is a solid or a semi-solid.

[1593] In some embodiments, the composition further comprises an anti-fungal agent. Suitable anti-fungal agents for use herein include, but are not limited to, fungicidal and fungistatic agents including terbinafine, itraconazole, micronazole nitrate, thiapendazole, tolnaftate, clotrimazole and griseofulvin. In some embodiments, the anti-fungal agent is a polyene anti-fungal agent, such as amphotericin-B, nystatin, and pimaricin.

[1594] In some embodiments, the composition does not contain any anti-fungal agent. In some embodiments, the composition contains broad spectrum antibiotics but not any anti-fungal agent. Such compositions that do not contain anti-fungal agents but contain broad spectrum antibiotics may be useful in detecting or quantifying fungi (e.g., yeast) in a sample.

[1595] In some embodiments, the composition does not contain any anti-fungal agent, any antibiotics or any anti-mammalian agent. Such compositions that do not selectively lyse mammalian cells may be useful in detecting or quantifying mammalian cells (e.g., cells from the GI tract) in a sample since many dyes have a higher affinity for mammalian as compared to bacteria or fungi cells. In some embodiments, the composition contains broad spectrum antibiotics and one or more anti-fungal agents. Such compositions that contain anti-fungal agents and broad spectrum antibiotics may be useful in detecting or quantifying mammalian cells (e.g., cells from the GI tract) in a sample. The detection or quantification of mammalian cells may be useful for determining cell turnover in a subject. High cell turnover is sometimes associated with a GI injury (e.g., lesion), the presence of a tumor(s), or radiation-induced colitis or radiation enteropathy.

[1596] In some embodiments, the composition further comprises an antibiotic agent as described herein. Such a composition may be useful in detecting or quantifying antibiotic-resistant strains of bacteria in a sample.

[1597] In certain embodiments, the composition comprises Triton X-100, deoxycholate, resazurin, and MgCl.sub.2. In some embodiments, the composition comprises Triton X-100, deoxycholate, resazurin, amphotericin-B and MgCl.sub.2. In some embodiments, the composition comprises 0.1 wt % or 0.05 wt % Triton X-100; 0.005 wt % deoxycholate; 10 mM resazurin; 2.5 mg/L amphotericin-B and 50 mM MgCl.sub.2. In some embodiments, the composition has a pH of 6.0.

[1598] In certain embodiments, the compositions are suitable for use in a kit or device, e.g., for detecting or quantifying viable bacterial cells in a sample. In some embodiments, such a device is an ingestible device for detecting or quantifying viable bacterial cells in vivo (e.g., in the GI tract).

[1599] FIG. 62 illustrates a nonlimiting example of a system for collecting, communicating and/or analyzing data about a subject, using an ingestible device as disclosed herein. For example, an ingestible device may be configured to communicate with an external base station. As an example, an ingestible device can have a communications unit that communicates with an external base station which itself has a communications unit. FIG. 62 illustrates exemplary implementation of such an ingestible device. As shown in FIG. 62, a subject ingests an ingestible device as disclosed herein. Certain data about the subject (e.g., based on a collected sample) and/or the location of the ingestible device in the GI tract of the subject is collected or otherwise available and provided to a mobile device, which then forwards the data via the internet and a server/data store to a physician's office computer. The information collected by the ingestible device is communicated to a receiver, such as, for example, a watch or other object worn by the subject. The information is then communicated from the receiver to the mobile device which then forwards the data via the internet and a server/data store to a physician's office computer. The physician is then able to analyze some or all of the data about the subject to provide recommendations, such as, for example, delivery a therapeutic agent. While FIG. 62 shows a particular approach to collecting and transferring data about a subject, the disclosure is not limited. As an example, one or more of the receiver, mobile device, internet, and/or server/data store can be excluded from the data communication channel. For example, a mobile device can be used as the receiver of the device data, e.g., by using a dongle. In such embodiments, the item worn by the subject need not be part of the communication chain. As another example, one or more of the items in the data communication channel can be replaced with an alternative item. For example, rather than be provided to a physician's office computer, data may be provided to a service provider network, such as a hospital network, an HMO network, or the like. In some embodiments, subject data may be collected and/or stored in one location (e.g., a server/data store) while device data may be collected and/or stored in a different location (e.g., a different server/data store).

Locations of Release

[1600] In some embodiments, the immune modulator is delivered at a location in the large intestine of the subject. In some embodiments, the location is in the proximal portion of the large intestine. In some embodiments, the location is in the distal portion of the large intestine.

[1601] In some embodiments, the immune modulator is delivered at a location in the ascending colon of the subject. In some embodiments, the location is in the proximal portion of the ascending colon. In some embodiments, the location is in the distal portion of the ascending colon.

[1602] In some embodiments, the immune modulator is delivered at a location in the cecum of the subject. In some embodiments, the location is in the proximal portion of the cecum. In some embodiments, the location is in the distal portion of the cecum.

[1603] In some embodiments, the immune modulator is delivered at a location in the sigmoid colon of the subject. In some embodiments, the location is in the proximal portion of the sigmoid colon. In some embodiments, the location is in the distal portion of the sigmoid colon.

[1604] In some embodiments, the immune modulator is delivered at a location in the transverse colon of the subject. In some embodiments, the location is in the proximal portion of the transverse colon. In some embodiments, the location is in the distal portion of the transverse colon.

[1605] In some embodiments, the immune modulator is delivered at a location in the descending colon of the subject. In some embodiments, the location is in the proximal portion of the descending colon. In some embodiments, the location is in the distal portion of the descending colon.

[1606] In some embodiments, the immune modulator is delivered at a location in the small intestine of the subject. In some embodiments, the location is in the proximal portion of the small intestine. In some embodiments, the location is in the distal portion of the small intestine.

[1607] In some embodiments, the immune modulator is delivered at a location in the duodenum of the subject. In some embodiments, the location is in the proximal portion of the duodenum. In some embodiments, the location is in the distal portion of the duodenum.

[1608] In some embodiments, the immune modulator is delivered at a location in the jejunum of the subject. In some embodiments, the location is in the proximal portion of the jejunum. In some embodiments, the location is in the distal portion of the jejunum.

[1609] In some embodiments, the immune modulator is delivered at a location in the duodenum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1610] In some embodiments, the immune modulator is delivered at a location in the proximal duodenum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1611] In some embodiments, the immune modulator is delivered at a location in the jejunum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1612] In some embodiments, the immune modulator is delivered at a location in the proximal portion of the jejunum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1613] In some embodiments, the immune modulator is delivered at a location in the distal portion of the jejunum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1614] In some embodiments, the immune modulator is delivered at a location in the ileum of the subject. In some embodiments, the location is in the proximal portion of the ileum. In some embodiments, the location is in the distal portion of the ileum.

[1615] In some embodiments, the immune modulator is delivered at a location in the ileum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1616] In some embodiments, the immune modulator is delivered at a location in the proximal portion of the ileum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1617] In some embodiments, the immune modulator is delivered at a location in the distal portion of the ileum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1618] In some embodiments, the immune modulator is delivered at a location in the cecum of the subject and is not delivered at other locations in the gastrointestinal tract.

[1619] In some embodiments, the location at which the immune modulator is delivered is proximate to the intended site of release of the immune modulator. In some embodiments, the immune modulator is delivered 150 cm or less from the one or more sites of disease. In some embodiments, the immune modulator is delivered 125 cm or less from the one or more sites of disease. In some embodiments, the immune modulator is delivered 100 cm or less from the one or more sites of disease. In some embodiments, the immune modulator is delivered 50 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 40 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 30 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 20 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 10 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 5 cm or less from the intended site of release. In some embodiments, the immune modulator is delivered 2 cm or less from the intended site of release. In some embodiments, the method further comprises using an ingestible device to deliver the immune modulator and using localization methods disclosed herein (e.g., such as discussed in Example 14 below) to determine the location of the ingestible device within the GI tract. In some embodiments, the method further comprises using an ingestible device to deliver the immune modulator and determining the period of time since the ingestible device was ingested to determine the location of the ingestible device within the GI tract. In some embodiments, the method further comprises imaging of the gastrointestinal tract. In some embodiments, imaging of the gastrointestinal tract comprises video imaging. In some embodiments, imaging of the gastrointestinal tract comprises thermal imaging. In some embodiments, imaging of the gastrointestinal tract comprises ultrasound imaging. In some embodiments, imaging of the gastrointestinal tract comprises Doppler imaging.

[1620] In some embodiments the method does not comprise releasing more than 20% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments the method does not comprise releasing more than 10% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments the method does not comprise releasing more than 5% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments the method does not comprise releasing more than 4% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments the method does not comprise releasing more than 3% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments the method does not comprise releasing more than 2% of the immune modulator at a location that is not proximate to the intended site of release.

[1621] In some embodiments the method comprises releasing at least 80% of the immune modulator at a location proximate to a site of disease. In some embodiments the method comprise releasing at least 90% of the immune modulator at a location proximate to a site of disease. In some embodiments the method comprises releasing at least 95% of the immune modulator at a location proximate to a site of disease. In some embodiments the method comprises releasing at least 96% of the immune modulator at a location proximate to a site of disease. In some embodiments the method comprises releasing at least 97% of the immune modulator at a location proximate to a site of disease. In some embodiments the method comprises releasing at least 98% of the immune modulator at a location proximate to a site of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 150 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 125 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 100 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 50 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 40 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 30 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 20 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 10 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 5 cm or less from the one or more sites of disease. In some embodiments, the at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% of the immune modulator is delivered 2 cm or less from the one or more sites of disease. In some embodiments, the method further comprises using an ingestible device to deliver the immune modulator and using localization methods disclosed herein (e.g., such as discussed in Example 14 below) to determine the location of the ingestible device within the GI tract (e.g., relative to the site of disease). In some embodiments, the method further comprises using an ingestible device to deliver the immune modulator and determining the period of time since the ingestible device was ingested to determine the location of the ingestible device within the GI tract (e.g., relative to the site of disease).

[1622] In some embodiments, the amount of immune modulator that is delivered is a Human Equivalent Dose.

[1623] In some embodiments the method comprises releasing the immune modulator at a location that is proximate to the intended site of release, wherein the immune modulator and, if applicable, any carriers, excipients or stabilizers admixed with the immune modulator, are substantially unchanged, at the time of release of the immune modulator at the location, relatively to the time of administration of the composition to the subject.

[1624] In some embodiments the method comprises releasing the immune modulator at a location that is proximate to the intended site of release, wherein the immune modulator and, if applicable, any carriers, excipients or stabilizers admixed with the immune modulator, are substantially unchanged by any physiological process (such as, but not limited to, degradation in the stomach), at the time of release of the immune modulator at the location, relatively to the time of administration of the composition to the subject.

[1625] In some embodiments, the immune modulator is delivered to the location by mucosal contact.

[1626] In some embodiments, a method of treatment disclosed herein includes determining the level of an immune modulator at the intended site of release or a location in the gastrointestinal tract of the subject that is proximate to the intended site of release. In some examples, a method of treatment as described herein can include determining the level of the immune modulator at the intended site of release or a location in the gastrointestinal tract of the subject that is proximate to the intended site of release within a time period of about 10 minutes to about 10 hours following administration of the device.

[1627] In some examples, a method of treatment disclosed herein includes determining the level of the anti-inflammatory at the intended site of release or a location in the gastrointestinal tract of the subject that is proximate to the intended site of release at a time point following administration of the device that is elevated as compared to a level of the immune modulator at the same site of release or location at substantially the same time point in a subject following systemic administration of an equal amount of the the immune modulator.

[1628] As used herein, GI tissue refers to tissue in the gastrointestinal (GI) tract, such as tissue in one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum, more particularly in the proximal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon, or in the distal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon. Accordingly, in some embodiments the immune modulator can penetrate the dudodenum tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the jejunum tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the ileum tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the cecum tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the ascending colon tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the transverse colon tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the descending colon tissue proximate to the intended site of release. In some embodiments the immune modulator can penetrate the sigmoid colon tissue proximate to the intended site of release. For example, an immune modulator can penetrate one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa.

[1629] In some examples, administration of an immune modulator using any of the compositions or devices described herein results in penetration (e.g., a detectable level of penetration) of GI tissue (e.g., one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa) within a time period of about 10 minutes to about 10 hours, about 10 minutes to about 9 hours, about 10 minutes to about 8 hours, about 10 minutes to about 7 hours, about 10 minutes to about 6 hours, about 10 minutes to about 5 hours, about 10 minutes to about 4.5 hours, about 10 minutes to about 4 hours, about 10 minutes to about 3.5 hours, about 10 minutes to about 3 hours, about 10 minutes to about 2.5 hours, about 10 minutes to about 2 hours, about 10 minutes to about 1.5 hours, about 10 minutes to about 1 hour, about 10 minutes to about 55 minutes, about 10 minutes to about 50 minutes, about 10 minutes to about 45 minutes, about 10 minutes to about 40 minutes, about 10 minutes to about 35 minutes, about 10 minutes to about 30 minutes, about 10 minutes to about 25 minutes, about 10 minutes to about 20 minutes, about 10 minutes to about 15 minutes, about 15 minutes to about 10 hours, about 15 minutes to about 9 hours, about 15 minutes to about 8 hours, about 15 minutes to about 7 hours, about 15 minutes to about 6 hours, about 15 minutes to about 5 hours, about 15 minutes to about 4.5 hours, about 15 minutes to about 4 hours, about 15 minutes to about 3.5 hours, about 15 minutes to about 3 hours, about 15 minutes to about 2.5 hours, about 15 minutes to about 2 hours, about 15 minutes to about 1.5 hours, about 15 minutes to about 1 hour, about 15 minutes to about 55 minutes, about 15 minutes to about 50 minutes, about 15 minutes to about 45 minutes, about 15 minutes to about 40 minutes, about 15 minutes to about 35 minutes, about 15 minutes to about 30 minutes, about 15 minutes to about 25 minutes, about 15 minutes to about 20 minutes, about 20 minutes to about 10 hours, about 20 minutes to about 9 hours, about 20 minutes to about 8 hours, about 20 minutes to about 7 hours, about 20 minutes to about 6 hours, about 20 minutes to about 5 hours, about 20 minutes to about 4.5 hours, about 20 minutes to about 4 hours, about 20 minutes to about 3.5 hours, about 20 minutes to about 3 hours, about 20 minutes to about 2.5 hours, about 20 minutes to about 2 hours, about 20 minutes to about 1.5 hours, about 20 minutes to about 1 hour, about 20 minutes to about 55 minutes, about 20 minutes to about 50 minutes, about 20 minutes to about 45 minutes, about 20 minutes to about 40 minutes, about 20 minutes to about 35 minutes, about 20 minutes to about 30 minutes, about 20 minutes to about 25 minutes, about 25 minutes to about 10 hours, about 25 minutes to about 9 hours, about 25 minutes to about 8 hours, about 25 minutes to about 7 hours, about 25 minutes to about 6 hours, about 25 minutes to about 5 hours, about 25 minutes to about 4.5 hours, about 25 minutes to about 4 hours, about 25 minutes to about 3.5 hours, about 25 minutes to about 3 hours, about 25 minutes to about 2.5 hours, about 25 minutes to about 2 hours, about 25 minutes to about 1.5 hours, about 25 minutes to about 1 hour, about 25 minutes to about 55 minutes, about 25 minutes to about 50 minutes, about 25 minutes to about 45 minutes, about 25 minutes to about 40 minutes, about 25 minutes to about 35 minutes, about 25 minutes to about 30 minutes, about 30 minutes to about 10 hours, about 30 minutes to about 9 hours, about 30 minutes to about 8 hours, about 30 minutes to about 7 hours, about 30 minutes to about 6 hours, about 30 minutes to about 5 hours, about 30 minutes to about 4.5 hours, about 30 minutes to about 4 hours, about 30 minutes to about 3.5 hours, about 30 minutes to about 3 hours, about 30 minutes to about 2.5 hours, about 30 minutes to about 2 hours, about 30 minutes to about 1.5 hours, about 30 minutes to about 1 hour, about 30 minutes to about 55 minutes, about 30 minutes to about 50 minutes, about 30 minutes to about 45 minutes, about 30 minutes to about 40 minutes, about 30 minutes to about 35 minutes, about 35 minutes to about 10 hours, about 35 minutes to about 9 hours, about 35 minutes to about 8 hours, about 35 minutes to about 7 hours, about 35 minutes to about 6 hours, about 35 minutes to about 5 hours, about 35 minutes to about 4.5 hours, about 35 minutes to about 4 hours, about 35 minutes to about 3.5 hours, about 35 minutes to about 3 hours, about 35 minutes to about 2.5 hours, about 35 minutes to about 2 hours, about 35 minutes to about 1.5 hours, about 35 minutes to about 1 hour, about 35 minutes to about 55 minutes, about 35 minutes to about 50 minutes, about 35 minutes to about 45 minutes, about 35 minutes to about 40 minutes, about 40 minutes to about 10 hours, about 40 minutes to about 9 hours, about 40 minutes to about 8 hours, about 40 minutes to about 7 hours, about 40 minutes to about 6 hours, about 40 minutes to about 5 hours, about 40 minutes to about 4.5 hours, about 40 minutes to about 4 hours, about 40 minutes to about 3.5 hours, about 40 minutes to about 3 hours, about 40 minutes to about 2.5 hours, about 40 minutes to about 2 hours, about 40 minutes to about 1.5 hours, about 40 minutes to about 1 hour, about 40 minutes to about 55 minutes, about 40 minutes to about 50 minutes, about 40 minutes to about 45 minutes, about 45 minutes to about 10 hours, about 45 minutes to about 9 hours, about 45 minutes to about 8 hours, about 45 minutes to about 7 hours, about 45 minutes to about 6 hours, about 45 minutes to about 5 hours, about 45 minutes to about 4.5 hours, about 45 minutes to about 4 hours, about 45 minutes to about 3.5 hours, about 45 minutes to about 3 hours, about 45 minutes to about 2.5 hours, about 45 minutes to about 2 hours, about 45 minutes to about 1.5 hours, about 45 minutes to about 1 hour, about 45 minutes to about 55 minutes, about 45 minutes to about 50 minutes, about 50 minutes to about 10 hours, about 50 minutes to about 9 hours, about 50 minutes to about 8 hours, about 50 minutes to about 7 hours, about 50 minutes to about 6 hours, about 50 minutes to about 5 hours, about 50 minutes to about 4.5 hours, about 50 minutes to about 4 hours, about 50 minutes to about 3.5 hours, about 50 minutes to about 3 hours, about 50 minutes to about 2.5 hours, about 50 minutes to about 2 hours, about 50 minutes to about 1.5 hours, about 50 minutes to about 1 hour, about 50 minutes to about 55 minutes, about 55 minutes to about 10 hours, about 55 minutes to about 9 hours, about 55 minutes to about 8 hours, about 55 minutes to about 7 hours, about 55 minutes to about 6 hours, about 55 minutes to about 5 hours, about 55 minutes to about 4.5 hours, about 55 minutes to about 4 hours, about 55 minutes to about 3.5 hours, about 55 minutes to about 3 hours, about 55 minutes to about 2.5 hours, about 55 minutes to about 2 hours, about 55 minutes to about 1.5 hours, about 55 minutes to about 1 hour, about 1 hour to about 10 hours, about 1 hour to about 9 hours, about 1 hour to about 8 hours, about 1 hour to about 7 hours, about 1 hour to about 6 hours, about 1 hour to about 5 hours, about 1 hour to about 4.5 hours, about 1 hour to about 4 hours, about 1 hour to about 3.5 hours, about 1 hour to about 3 hours, about 1 hour to about 2.5 hours, about 1 hour to about 2 hours, about 1 hour to about 1.5 hours, about 1.5 hours to about 10 hours, about 1.5 hours to about 9 hours, about 1.5 hours to about 8 hours, about 1.5 hours to about 7 hours, about 1.5 hours to about 6 hours, about 1.5 hours to about 5 hours, about 1.5 hours to about 4.5 hours, about 1.5 hours to about 4 hours, about 1.5 hours to about 3.5 hours, about 1.5 hours to about 3 hours, about 1.5 hours to about 2.5 hours, about 1.5 hours to about 2 hours, about 2 hours to about 10 hours, about 2 hours to about 9 hours, about 2 hours to about 8 hours, about 2 hours to about 7 hours, about 2 hours to about 6 hours, about 2 hours to about 5 hours, about 2 hours to about 4.5 hours, about 2 hours to about 4 hours, about 2 hours to about 3.5 hours, about 2 hours to about 3 hours, about 2 hours to about 2.5 hours, about 2.5 hours to about 10 hours, about 2.5 hours to about 9 hours, about 2.5 hours to about 8 hours, about 2.5 hours to about 7 hours, about 2.5 hours to about 6 hours, about 2.5 hours to about 5 hours, about 2.5 hours to about 4.5 hours, about 2.5 hours to about 4 hours, about 2.5 hours to about 3.5 hours, about 2.5 hours to about 3 hours, about 3 hours to about 10 hours, about 3 hours to about 9 hours, about 3 hours to about 8 hours, about 3 hours to about 7 hours, about 3 hours to about 6 hours, about 3 hours to about 5 hours, about 3 hours to about 4.5 hours, about 3 hours to about 4 hours, about 3 hours to about 3.5 hours, about 3.5 hours to about 10 hours, about 3.5 hours to about 9 hours, about 3.5 hours to about 8 hours, about 3.5 hours to about 7 hours, about 3.5 hours to about 6 hours, about 3.5 hours to about 5 hours, about 3.5 hours to about 4.5 hours, about 3.5 hours to about 4 hours, about 4 hours to about 10 hours, about 4 hours to about 9 hours, about 4 hours to about 8 hours, about 4 hours to about 7 hours, about 4 hours to about 6 hours, about 4 hours to about 5 hours, about 4 hours to about 4.5 hours, about 4.5 hours to about 10 hours, about 4.5 hours to about 9 hours, about 4.5 hours to about 8 hours, about 4.5 hours to about 7 hours, about 4.5 hours to about 6 hours, about 4.5 hours to about 5 hours, about 5 hours to about 10 hours, about 5 hours to about 9 hours, about 5 hours to about 8 hours, about 5 hours to about 7 hours, about 5 hours to about 6 hours, about 6 hours to about 10 hours, about 6 hours to about 9 hours, about 6 hours to about 8 hours, about 6 hours to about 7 hours, about 7 hours to about 10 hours, about 7 hours to about 9 hours, about 7 hours to about 8 hours, about 8 hours to about 10 hours, about 8 hours to about 9 hours, or about 9 hours to about 10 hours. Penetration of GI tissue by an immune modulator can be detected by administering a labeled immune modulator, and performing imaging on the subject (e.g., ultrasound, computed tomography, or magnetic resonance imaging). For example, the label can be a radioisotope, a heavy metal, a fluorophore, or a luminescent agent (e.g., any suitable radioisotopes, heavy metals, fluorophores, or luminescent agents used for imaging known in the art).

[1630] In some embodiments, administration of an immune modulator can provide for treatment (e.g., a reduction in the number, severity, and/or duration of one or more symptoms of any of the disorders described herein in a subject) for a time period of between about 1 hour to about 30 days, about 1 hour to about 28 days, about 1 hour to about 26 days, about 1 hour to about 24 days, about 1 hour to about 22 days, about 1 hour to about 20 days, about 1 hour to about 18 days, about 1 hour to about 16 days, about 1 hour to about 14 days, about 1 hour to about 12 days, about 1 hour to about 10 days, about 1 hour to about 8 days, about 1 hour to about 6 days, about 1 hour to about 5 days, about 1 hour to about 4 days, about 1 hour to about 3 days, about 1 hour to about 2 days, about 1 hour to about 1 day, about 1 hour to about 12 hours, about 1 hour to about 6 hours, about 1 hour to about 3 hours, about 3 hours to about 30 days, about 3 hours to about 28 days, about 3 hours to about 26 days, about 3 hours to about 24 days, about 3 hours to about 22 days, about 3 hours to about 20 days, about 3 hours to about 18 days, about 3 hours to about 16 days, about 3 hours to about 14 days, about 3 hours to about 12 days, about 3 hours to about 10 days, about 3 hours to about 8 days, about 3 hours to about 6 days, about 3 hours to about 5 days, about 3 hours to about 4 days, about 3 hours to about 3 days, about 3 hours to about 2 days, about 3 hours to about 1 day, about 3 hours to about 12 hours, about 3 hours to about 6 hours, about 6 hours to about 30 days, about 6 hours to about 28 days, about 6 hours to about 26 days, about 6 hours to about 24 days, about 6 hours to about 22 days, about 6 hours to about 20 days, about 6 hours to about 18 days, about 6 hours to about 16 days, about 6 hours to about 14 days, about 6 hours to about 12 days, about 6 hours to about 10 days, about 6 hours to about 8 days, about 6 hours to about 6 days, about 6 hours to about 5 days, about 6 hours to about 4 days, about 6 hours to about 3 days, about 6 hours to about 2 days, about 6 hours to about 1 day, about 6 hours to about 12 hours, about 12 hours to about 30 days, about 12 hours to about 28 days, about 12 hours to about 26 days, about 12 hours to about 24 days, about 12 hours to about 22 days, about 12 hours to about 20 days, about 12 hours to about 18 days, about 12 hours to about 16 days, about 12 hours to about 14 days, about 12 hours to about 12 days, about 12 hours to about 10 days, about 12 hours to about 8 days, about 12 hours to about 6 days, about 12 hours to about 5 days, about 12 hours to about 4 days, about 12 hours to about 3 days, about 12 hours to about 2 days, about 12 hours to about 1 day, about 1 day to about 30 days, about 1 day to about 28 days, about 1 day to about 26 days, about 1 day to about 24 days, about 1 day to about 22 days, about 1 day to about 20 days, about 1 day to about 18 days, about 1 day to about 16 days, about 1 day to about 14 days, about 1 day to about 12 days, about 1 day to about 10 days, about 1 day to about 8 days, about 1 day to about 6 days, about 1 day to about 5 days, about 1 day to about 4 days, about 1 day to about 3 days, about 1 day to about 2 days, about 2 days to about 30 days, about 2 days to about 28 days, about 2 days to about 26 days, about 2 days to about 24 days, about 2 days to about 22 days, about 2 days to about 20 days, about 2 days to about 18 days, about 2 days to about 16 days, about 2 days to about 14 days, about 2 days to about 12 days, about 2 days to about 10 days, about 2 days to about 8 days, about 2 days to about 6 days, about 2 days to about 5 days, about 2 days to about 4 days, about 2 days to about 3 days, about 3 days to about 30 days, about 3 days to about 28 days, about 3 days to about 26 days, about 3 days to about 24 days, about 3 days to about 22 days, about 3 days to about 20 days, about 3 days to about 18 days, about 3 days to about 16 days, about 3 days to about 14 days, about 3 days to about 12 days, about 3 days to about 10 days, about 3 days to about 8 days, about 3 days to about 6 days, about 3 days to about 5 days, about 3 days to about 4 days, about 4 days to about 30 days, about 4 days to about 28 days, about 4 days to about 26 days, about 4 days to about 24 days, about 4 days to about 22 days, about 4 days to about 20 days, about 4 days to about 18 days, about 4 days to about 16 days, about 4 days to about 14 days, about 4 days to about 12 days, about 4 days to about 10 days, about 4 days to about 8 days, about 4 days to about 6 days, about 4 days to about 5 days, about 5 days to about 30 days, about 5 days to about 28 days, about 5 days to about 26 days, about 5 days to about 24 days, about 5 days to about 22 days, about 5 days to about 20 days, about 5 days to about 18 days, about 5 days to about 16 days, about 5 days to about 14 days, about 5 days to about 12 days, about 5 days to about 10 days, about 5 days to about 8 days, about 5 days to about 6 days, about 6 days to about 30 days, about 6 days to about 28 days, about 6 days to about 26 days, about 6 days to about 24 days, about 6 days to about 22 days, about 6 days to about 20 days, about 6 days to about 18 days, about 6 days to about 16 days, about 6 days to about 14 days, about 6 days to about 12 days, about 6 days to about 10 days, about 6 days to about 8 days, about 8 days to about 30 days, about 8 days to about 28 days, about 8 days to about 26 days, about 8 days to about 24 days, about 8 days to about 22 days, about 8 days to about 20 days, about 8 days to about 18 days, about 8 days to about 16 days, about 8 days to about 14 days, about 8 days to about 12 days, about 8 days to about 10 days, about 10 days to about 30 days, about 10 days to about 28 days, about 10 days to about 26 days, about 10 days to about 24 days, about 10 days to about 22 days, about 10 days to about 20 days, about 10 days to about 18 days, about 10 days to about 16 days, about 10 days to about 14 days, about 10 days to about 12 days, about 12 days to about 30 days, about 12 days to about 28 days, about 12 days to about 26 days, about 12 days to about 24 days, about 12 days to about 22 days, about 12 days to about 20 days, about 12 days to about 18 days, about 12 days to about 16 days, about 12 days to about 14 days, about 14 days to about 30 days, about 14 days to about 28 days, about 14 days to about 26 days, about 14 days to about 24 days, about 14 days to about 22 days, about 14 days to about 20 days, about 14 days to about 18 days, about 14 days to about 16 days, about 16 days to about 30 days, about 16 days to about 28 days, about 16 days to about 26 days, about 16 days to about 24 days, about 16 days to about 22 days, about 16 days to about 20 days, about 16 days to about 18 days, about 18 days to about 30 days, about 18 days to about 28 days, about 18 days to about 26 days, about 18 days to about 24 days, about 18 days to about 22 days, about 18 days to about 20 days, about 20 days to about 30 days, about 20 days to about 28 days, about 20 days to about 26 days, about 20 days to about 24 days, about 20 days to about 22 days, about 22 days to about 30 days, about 22 days to about 28 days, about 22 days to about 26 days, about 22 days to about 24 days, about 24 days to about 30 days, about 24 days to about 28 days, about 24 days to about 26 days, about 26 days to about 30 days, about 26 days to about 28 days, or about 28 days to about 30 days in a subject following first administration of an immune modulator using any of the compositions or devices described herein. Non-limiting examples of symptoms of a disease described herein are described below.

[1631] For example, treatment can result in a decrease (e.g., about 1% to about 99% decrease, about 1% to about 95% decrease, about 1% to about 90% decrease, about 1% to about 85% decrease, about 1% to about 80% decrease, about 1% to about 75% decrease, about 1% to about 70% decrease, about 1% to about 65% decrease, about 1% to about 60% decrease, about 1% to about 55% decrease, about 1% to about 50% decrease, about 1% to about 45% decrease, about 1% to about 40% decrease, about 1% to about 35% decrease, about 1% to about 30% decrease, about 1% to about 25% decrease, about 1% to about 20% decrease, about 1% to about 15% decrease, about 1% to about 10% decrease, about 1% to about 5% decrease, about 5% to about 99% decrease, about 5% to about 95% decrease, about 5% to about 90% decrease, about 5% to about 85% decrease, about 5% to about 80% decrease, about 5% to about 75% decrease, about 5% to about 70% decrease, about 5% to about 65% decrease, about 5% to about 60% decrease, about 5% to about 55% decrease, about 5% to about 50% decrease, about 5% to about 45% decrease, about 5% to about 40% decrease, about 5% to about 35% decrease, about 5% to about 30% decrease, about 5% to about 25% decrease, about 5% to about 20% decrease, about 5% to about 15% decrease, about 5% to about 10% decrease, about 10% to about 99% decrease, about 10% to about 95% decrease, about 10% to about 90% decrease, about 10% to about 85% decrease, about 10% to about 80% decrease, about 10% to about 75% decrease, about 10% to about 70% decrease, about 10% to about 65% decrease, about 10% to about 60% decrease, about 10% to about 55% decrease, about 10% to about 50% decrease, about 10% to about 45% decrease, about 10% to about 40% decrease, about 10% to about 35% decrease, about 10% to about 30% decrease, about 10% to about 25% decrease, about 10% to about 20% decrease, about 10% to about 15% decrease, about 15% to about 99% decrease, about 15% to about 95% decrease, about 15% to about 90% decrease, about 15% to about 85% decrease, about 15% to about 80% decrease, about 15% to about 75% decrease, about 15% to about 70% decrease, about 15% to about 65% decrease, about 15% to about 60% decrease, about 15% to about 55% decrease, about 15% to about 50% decrease, about 15% to about 45% decrease, about 15% to about 40% decrease, about 15% to about 35% decrease, about 15% to about 30% decrease, about 15% to about 25% decrease, about 15% to about 20% decrease, about 20% to about 99% decrease, about 20% to about 95% decrease, about 20% to about 90% decrease, about 20% to about 85% decrease, about 20% to about 80% decrease, about 20% to about 75% decrease, about 20% to about 70% decrease, about 20% to about 65% decrease, about 20% to about 60% decrease, about 20% to about 55% decrease, about 20% to about 50% decrease, about 20% to about 45% decrease, about 20% to about 40% decrease, about 20% to about 35% decrease, about 20% to about 30% decrease, about 20% to about 25% decrease, about 25% to about 99% decrease, about 25% to about 95% decrease, about 25% to about 90% decrease, about 25% to about 85% decrease, about 25% to about 80% decrease, about 25% to about 75% decrease, about 25% to about 70% decrease, about 25% to about 65% decrease, about 25% to about 60% decrease, about 25% to about 55% decrease, about 25% to about 50% decrease, about 25% to about 45% decrease, about 25% to about 40% decrease, about 25% to about 35% decrease, about 25% to about 30% decrease, about 30% to about 99% decrease, about 30% to about 95% decrease, about 30% to about 90% decrease, about 30% to about 85% decrease, about 30% to about 80% decrease, about 30% to about 75% decrease, about 30% to about 70% decrease, about 30% to about 65% decrease, about 30% to about 60% decrease, about 30% to about 55% decrease, about 30% to about 50% decrease, about 30% to about 45% decrease, about 30% to about 40% decrease, about 30% to about 35% decrease, about 35% to about 99% decrease, about 35% to about 95% decrease, about 35% to about 90% decrease, about 35% to about 85% decrease, about 35% to about 80% decrease, about 35% to about 75% decrease, about 35% to about 70% decrease, about 35% to about 65% decrease, about 35% to about 60% decrease, about 35% to about 55% decrease, about 35% to about 50% decrease, about 35% to about 45% decrease, about 35% to about 40% decrease, about 40% to about 99% decrease, about 40% to about 95% decrease, about 40% to about 90% decrease, about 40% to about 85% decrease, about 40% to about 80% decrease, about 40% to about 75% decrease, about 40% to about 70% decrease, about 40% to about 65% decrease, about 40% to about 60% decrease, about 40% to about 55% decrease, about 40% to about 50% decrease, about 40% to about 45% decrease, about 45% to about 99% decrease, about 45% to about 95% decrease, about 45% to about 90% decrease, about 45% to about 85% decrease, about 45% to about 80% decrease, about 45% to about 75% decrease, about 45% to about 70% decrease, about 45% to about 65% decrease, about 45% to about 60% decrease, about 45% to about 55% decrease, about 45% to about 50% decrease, about 50% to about 99% decrease, about 50% to about 95% decrease, about 50% to about 90% decrease, about 50% to about 85% decrease, about 50% to about 80% decrease, about 50% to about 75% decrease, about 50% to about 70% decrease, about 50% to about 65% decrease, about 50% to about 60% decrease, about 50% to about 55% decrease, about 55% to about 99% decrease, about 55% to about 95% decrease, about 55% to about 90% decrease, about 55% to about 85% decrease, about 55% to about 80% decrease, about 55% to about 75% decrease, about 55% to about 70% decrease, about 55% to about 65% decrease, about 55% to about 60% decrease, about 60% to about 99% decrease, about 60% to about 95% decrease, about 60% to about 90% decrease, about 60% to about 85% decrease, about 60% to about 80% decrease, about 60% to about 75% decrease, about 60% to about 70% decrease, about 60% to about 65% decrease, about 65% to about 99% decrease, about 65% to about 95% decrease, about 65% to about 90% decrease, about 65% to about 85% decrease, about 65% to about 80% decrease, about 65% to about 75% decrease, about 65% to about 70% decrease, about 70% to about 99% decrease, about 70% to about 95% decrease, about 70% to about 90% decrease, about 70% to about 85% decrease, about 70% to about 80% decrease, about 70% to about 75% decrease, about 75% to about 99% decrease, about 75% to about 95% decrease, about 75% to about 90% decrease, about 75% to about 85% decrease, about 75% to about 80% decrease, about 80% to about 99% decrease, about 80% to about 95% decrease, about 80% to about 90% decrease, about 80% to about 85% decrease, about 85% to about 99% decrease, about 85% to about 95% decrease, about 85% to about 90% decrease, about 90% to about 99% decrease, about 90% to about 95% decrease, or about 95% to about 99% decrease) in one or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) of: the level of interferon- in GI tissue, the level of IL-1 in GI tissue, the level of IL-6 in GI tissue, the level of IL-22 in GI tissue, the level of IL-17A in the GI tissue, the level of TNF in GI tissue, the level of IL-2 in GI tissue, the number of Th memory cells in Peyer's patches, and the number of Th memory cells in mesentery lymph nodes, in a subject (e.g., as compared to the level in the subject prior to treatment or compared to a subject or population of subjects having a similar disease but receiving a placebo or a different treatment) (e.g., for a time period of between about 1 hour to about 30 days (e.g., or any of the subranges herein) following the first administration of an immune modulator using any of the compositions or devices described herein. Exemplary methods for determining the endoscopy score are described herein and other methods for determining the endoscopy score are known in the art. Exemplary methods for determining the levels of interferon-, IL-1, IL-6, IL-22, IL-17A, TNF, and IL-2 are described herein. Additional methods for determining the levels of these cytokines are known in the art. Exemplary methods for determining the number of Th memory cells in Peyer's patches and mesentery lymph nodes are described herein. Additional methods for determining the number of Th memory cells in Peyer's patches and mesentery lymph nodes are known in the art.

[1632] In some examples, treatment can result in an increase (e.g., about 1% to about 500% increase, about 1% to about 400% increase, about 1% to about 300% increase, about 1% to about 200% increase, about 1% to about 150% increase, about 1% to about 100% increase, about 1% to about 90% increase, about 1% to about 80% increase, about 1% to about 70% increase, about 1% to about 60% increase, about 1% to about 50% increase, about 1% to about 40% increase, about 1% to about 30% increase, about 1% to about 20% increase, about 1% to about 10% increase, a 10% to about 500% increase, about 10% to about 400% increase, about 10% to about 300% increase, about 10% to about 200% increase, about 10% to about 150% increase, about 10% to about 100% increase, about 10% to about 90% increase, about 10% to about 80% increase, about 10% to about 70% increase, about 10% to about 60% increase, about 10% to about 50% increase, about 10% to about 40% increase, about 10% to about 30% increase, about 10% to about 20% increase, about 20% to about 500% increase, about 20% to about 400% increase, about 20% to about 300% increase, about 20% to about 200% increase, about 20% to about 150% increase, about 20% to about 100% increase, about 20% to about 90% increase, about 20% to about 80% increase, about 20% to about 70% increase, about 20% to about 60% increase, about 20% to about 50% increase, about 20% to about 40% increase, about 20% to about 30% increase, about 30% to about 500% increase, about 30% to about 400% increase, about 30% to about 300% increase, about 30% to about 200% increase, about 30% to about 150% increase, about 30% to about 100% increase, about 30% to about 90% increase, about 30% to about 80% increase, about 30% to about 70% increase, about 30% to about 60% increase, about 30% to about 50% increase, about 30% to about 40% increase, about 40% to about 500% increase, about 40% to about 400% increase, about 40% to about 300% increase, about 40% to about 200% increase, about 40% to about 150% increase, about 40% to about 100% increase, about 40% to about 90% increase, about 40% to about 80% increase, about 40% to about 70% increase, about 40% to about 60% increase, about 40% to about 50% increase, about 50% to about 500% increase, about 50% to about 400% increase, about 50% to about 300% increase, about 50% to about 200% increase, about 50% to about 150% increase, about 50% to about 100% increase, about 50% to about 90% increase, about 50% to about 80% increase, about 50% to about 70% increase, about 50% to about 60% increase, about 60% to about 500% increase, about 60% to about 400% increase, about 60% to about 300% increase, about 60% to about 200% increase, about 60% to about 150% increase, about 60% to about 100% increase, about 60% to about 90% increase, about 60% to about 80% increase, about 60% to about 70% increase, about 70% to about 500% increase, about 70% to about 400% increase, about 70% to about 300% increase, about 70% to about 200% increase, about 70% to about 150% increase, about 70% to about 100% increase, about 70% to about 90% increase, about 70% to about 80% increase, about 80% to about 500% increase, about 80% to about 400% increase, about 80% to about 300% increase, about 80% to about 200% increase, about 80% to about 150% increase, about 80% to about 100% increase, about 80% to about 90% increase, about 90% to about 500% increase, about 90% to about 400% increase, about 90% to about 300% increase, about 90% to about 200% increase, about 90% to about 150% increase, about 90% to about 100% increase, about 100% to about 500% increase, about 100% to about 400% increase, about 100% to about 300% increase, about 100% to about 200% increase, about 100% to about 150% increase, about 150% to about 500% increase, about 150% to about 400% increase, about 150% to about 300% increase, about 150% to about 200% increase, about 200% to about 500% increase, about 200% to about 400% increase, about 200% to about 300% increase, about 300% to about 500% increase, about 300% to about 400% increase, or about 400% to about 500% increase) in one or both of stool consistency score and weight of a subject (e.g., as compared to the level in the subject prior to treatment or compared to a subject or population of subjects having a similar disease but receiving a placebo or a different treatment) (e.g., for a time period of between about 1 hour to about 30 days (e.g., or any of the subranges herein) following the first administration of an immune modulator using any of the compositions or devices described herein. Exemplary methods for determining stool consistency score are described herein. Additional methods for determining a stool consistency score are known in the art.

[1633] In some embodiments, administration of an immune modulator using any of the devices or compositions described herein can result in a ratio of GI tissue concentration of the immune modulator to the blood, serum, or plasma concentration of the immune modulator that is higher than the same ratio when the immune modulator is administered by traditional means (e.g., systemically or orally). Examples of a ratio of GI tissue concentration of the immune modulator to the blood, serum, or plasma concentration of the immune modulator include about 2 to about 600, about 2 to about 580, about 2 to about 560, about 2 to about 540, about 2 to about 520, about 2 to about 500, about 2 to about 480, about 2 to about 460, about 4 to about 440, about 2 to about 420, about 2 to about 400, about 2 to about 380, about 2 to about 360, about 2 to about 340, about 2 to about 320, about 2 to about 300, about 2 to about 280, about 2 to about 260, about 2 to about 240, about 2 to about 220, about 2 to about 200, about 2 to about 190, about 2 to about 180, about 2 to about 170, about 2 to about 160, about 2 to about 150, about 2 to about 140, about 2 to about 130, about 2 to about 120, about 2 to about 110, about 2 to about 100, about 2 to about 90, about 2 to about 80, about 2 to about 70, about 2 to about 60, about 2 to about 50, about 2 to about 40, about 2 to about 30, about 2 to about 20, about 2 to about 15, about 2 to about 10, about 2 to about 5, about 5 to about 600, about 5 to about 580, about 5 to about 560, about 5 to about 540, about 5 to about 520, about 5 to about 500, about 5 to about 480, about 5 to about 460, about 5 to about 440, about 5 to about 420, about 5 to about 400, about 5 to about 380, about 5 to about 360, about 5 to about 340, about 5 to about 320, about 5 to about 300, about 5 to about 280, about 5 to about 260, about 5 to about 240, about 5 to about 220, about 5 to about 200, about 5 to about 190, about 5 to about 180, about 5 to about 170, about 5 to about 160, about 5 to about 150, about 5 to about 140, about 5 to about 130, about 5 to about 120, about 5 to about 110, about 5 to about 100, about 5 to about 90, about 5 to about 80, about 5 to about 70, about 5 to about 60, about 5 to about 50, about 5 to about 40, about 5 to about 30, about 5 to about 20, about 5 to about 15, about 5 to about 10, about 10 to about 600, about 10 to about 580, about 10 to about 560, about 10 to about 540, about 10 to about 520, about 10 to about 500, about 10 to about 480, about 10 to about 460, about 10 to about 440, about 10 to about 420, about 10 to about 400, about 10 to about 380, about 10 to about 360, about 10 to about 340, about 10 to about 320, about 10 to about 300, about 10 to about 280, about 10 to about 260, about 10 to about 240, about 10 to about 220, about 10 to about 200, about 10 to about 190, about 10 to about 180, about 10 to about 170, about 10 to about 160, about 10 to about 150, about 10 to about 140, about 10 to about 130, about 10 to about 120, about 10 to about 110, about 10 to about 100, about 10 to about 90, about 10 to about 80, about 10 to about 70, about 10 to about 60, about 10 to about 50, about 10 to about 40, about 10 to about 30, about 10 to about 20, about 10 to about 15, about 15 to about 600, about 15 to about 580, about 15 to about 560, about 15 to about 540, about 15 to about 520, about 15 to about 500, about 15 to about 480, about 15 to about 460, about 15 to about 440, about 15 to about 420, about 15 to about 400, about 15 to about 380, about 15 to about 360, about 15 to about 340, about 15 to about 320, about 15 to about 300, about 15 to about 280, about 15 to about 260, about 15 to about 240, about 15 to about 220, about 15 to about 200, about 15 to about 190, about 15 to about 180, about 15 to about 170, about 15 to about 160, about 15 to about 150, about 15 to about 140, about 15 to about 130, about 15 to about 120, about 15 to about 110, about 15 to about 100, about 15 to about 90, about 15 to about 80, about 15 to about 70, about 15 to about 60, about 15 to about 50, about 15 to about 40, about 15 to about 30, about 15 to about 20, about 20 to about 600, about 20 to about 580, about 20 to about 560, about 20 to about 540, about 20 to about 520, about 20 to about 500, about 20 to about 480, about 20 to about 460, about 20 to about 440, about 20 to about 420, about 20 to about 400, about 20 to about 380, about 20 to about 360, about 20 to about 340, about 20 to about 320, about 20 to about 300, about 20 to about 280, about 20 to about 260, about 20 to about 240, about 20 to about 220, about 20 to about 200, about 20 to about 190, about 20 to about 180, about 20 to about 170, about 20 to about 160, about 20 to about 150, about 20 to about 140, about 20 to about 130, about 20 to about 120, about 20 to about 110, about 20 to about 100, about 20 to about 90, about 20 to about 80, about 20 to about 70, about 20 to about 60, about 20 to about 50, about 20 to about 40, about 20 to about 30, about 30 to about 600, about 30 to about 580, about 30 to about 560, about 30 to about 540, about 30 to about 520, about 30 to about 500, about 30 to about 480, about 30 to about 460, about 30 to about 440, about 30 to about 420, about 30 to about 400, about 30 to about 380, about 30 to about 360, about 30 to about 340, about 30 to about 320, about 30 to about 300, about 30 to about 280, about 30 to about 260, about 30 to about 240, about 30 to about 220, about 30 to about 200, about 30 to about 190, about 30 to about 180, about 30 to about 170, about 30 to about 160, about 30 to about 150, about 30 to about 140, about 30 to about 130, about 30 to about 120, about 30 to about 110, about 30 to about 100, about 30 to about 90, about 30 to about 80, about 30 to about 70, about 30 to about 60, about 30 to about 50, about 30 to about 40, about 40 to about 600, about 40 to about 580, about 40 to about 560, about 40 to about 540, about 40 to about 520, about 40 to about 500, about 40 to about 480, about 40 to about 460, about 40 to about 440, about 40 to about 420, about 40 to about 400, about 40 to about 380, about 40 to about 360, about 40 to about 340, about 40 to about 320, about 40 to about 300, about 40 to about 280, about 40 to about 260, about 40 to about 240, about 40 to about 220, about 40 to about 200, about 40 to about 190, about 40 to about 180, about 40 to about 170, about 40 to about 160, about 40 to about 150, about 40 to about 140, about 40 to about 130, about 40 to about 120, about 40 to about 110, about 40 to about 100, about 40 to about 90, about 40 to about 80, about 40 to about 70, about 40 to about 60, about 40 to about 50, about 50 to about 600, about 50 to about 580, about 50 to about 560, about 50 to about 540, about 50 to about 520, about 50 to about 500, about 50 to about 480, about 50 to about 460, about 50 to about 440, about 50 to about 420, about 50 to about 400, about 50 to about 380, about 50 to about 360, about 50 to about 340, about 50 to about 320, about 50 to about 300, about 50 to about 280, about 50 to about 260, about 50 to about 240, about 50 to about 220, about 50 to about 200, about 50 to about 190, about 50 to about 180, about 50 to about 170, about 50 to about 160, about 50 to about 150, about 50 to about 140, about 50 to about 130, about 50 to about 120, about 50 to about 110, about 50 to about 100, about 50 to about 90, about 50 to about 80, about 50 to about 70, about 50 to about 60, about 60 to about 600, about 60 to about 580, about 60 to about 560, about 60 to about 540, about 60 to about 520, about 60 to about 500, about 60 to about 480, about 60 to about 460, about 60 to about 440, about 60 to about 420, about 60 to about 400, about 60 to about 380, about 60 to about 360, about 60 to about 340, about 60 to about 320, about 60 to about 300, about 60 to about 280, about 60 to about 260, about 60 to about 240, about 60 to about 220, about 60 to about 200, about 60 to about 190, about 60 to about 180, about 60 to about 170, about 60 to about 160, about 60 to about 150, about 60 to about 140, about 60 to about 130, about 60 to about 120, about 60 to about 110, about 60 to about 100, about 60 to about 90, about 60 to about 80, about 60 to about 70, about 70 to about 600, about 70 to about 580, about 70 to about 560, about 70 to about 540, about 70 to about 520, about 70 to about 500, about 70 to about 480, about 70 to about 460, about 70 to about 440, about 70 to about 420, about 70 to about 400, about 70 to about 380, about 70 to about 360, about 70 to about 340, about 70 to about 320, about 70 to about 300, about 70 to about 280, about 70 to about 260, about 70 to about 240, about 70 to about 220, about 70 to about 200, about 70 to about 190, about 70 to about 180, about 70 to about 170, about 70 to about 160, about 70 to about 150, about 70 to about 140, about 70 to about 130, about 70 to about 120, about 70 to about 110, about 70 to about 100, about 70 to about 90, about 70 to about 80, about 80 to about 600, about 80 to about 580, about 80 to about 560, about 80 to about 540, about 80 to about 520, about 80 to about 500, about 80 to about 480, about 80 to about 460, about 80 to about 440, about 80 to about 420, about 80 to about 400, about 80 to about 380, about 80 to about 360, about 80 to about 340, about 80 to about 320, about 80 to about 300, about 80 to about 280, about 80 to about 260, about 80 to about 240, about 80 to about 220, about 80 to about 200, about 80 to about 190, about 80 to about 180, about 80 to about 170, about 80 to about 160, about 80 to about 150, about 80 to about 140, about 80 to about 130, about 80 to about 120, about 80 to about 110, about 80 to about 100, about 80 to about 90, about 90 to about 600, about 90 to about 580, about 90 to about 560, about 90 to about 540, about 90 to about 520, about 90 to about 500, about 90 to about 480, about 90 to about 460, about 90 to about 440, about 90 to about 420, about 90 to about 400, about 90 to about 380, about 90 to about 360, about 90 to about 340, about 90 to about 320, about 90 to about 300, about 90 to about 280, about 90 to about 260, about 90 to about 240, about 90 to about 220, about 90 to about 200, about 90 to about 190, about 90 to about 180, about 90 to about 170, about 90 to about 160, about 90 to about 150, about 90 to about 140, about 90 to about 130, about 90 to about 120, about 90 to about 110, about 90 to about 100, about 100 to about 600, about 100 to about 580, about 100 to about 560, about 100 to about 540, about 100 to about 520, about 100 to about 500, about 100 to about 480, about 100 to about 460, about 100 to about 440, about 100 to about 420, about 100 to about 400, about 100 to about 380, about 100 to about 360, about 100 to about 340, about 100 to about 320, about 100 to about 300, about 100 to about 280, about 100 to about 260, about 100 to about 240, about 100 to about 220, about 100 to about 200, about 100 to about 190, about 100 to about 180, about 100 to about 170, about 100 to about 160, about 100 to about 150, about 100 to about 140, about 100 to about 130, about 100 to about 120, about 100 to about 110, about 110 to about 600, about 110 to about 580, about 110 to about 560, about 110 to about 540, about 110 to about 520, about 110 to about 500, about 110 to about 480, about 110 to about 460, about 110 to about 440, about 110 to about 420, about 110 to about 400, about 110 to about 380, about 110 to about 360, about 110 to about 340, about 110 to about 320, about 110 to about 300, about 110 to about 280, about 110 to about 260, about 110 to about 240, about 110 to about 220, about 110 to about 200, about 110 to about 190, about 110 to about 180, about 110 to about 170, about 110 to about 160, about 110 to about 150, about 110 to about 140, about 110 to about 130, about 110 to about 120, about 120 to about 600, about 120 to about 580, about 120 to about 560, about 120 to about 540, about 120 to about 520, about 120 to about 500, about 120 to about 480, about 120 to about 460, about 120 to about 440, about 120 to about 420, about 120 to about 400, about 120 to about 380, about 120 to about 360, about 120 to about 340, about 120 to about 320, about 120 to about 300, about 120 to about 280, about 120 to about 260, about 120 to about 240, about 120 to about 220, about 120 to about 200, about 120 to about 190, about 120 to about 180, about 120 to about 170, about 120 to about 160, about 120 to about 150, about 120 to about 140, about 120 to about 130, about 130 to about 600, about 130 to about 580, about 130 to about 560, about 130 to about 540, about 130 to about 520, about 130 to about 500, about 130 to about 480, about 130 to about 460, about 130 to about 440, about 130 to about 420, about 130 to about 400, about 130 to about 380, about 130 to about 360, about 130 to about 340, about 130 to about 320, about 130 to about 300, about 130 to about 280, about 130 to about 260, about 130 to about 240, about 130 to about 220, about 130 to about 200, about 130 to about 190, about 130 to about 180, about 130 to about 170, about 130 to about 160, about 130 to about 150, about 130 to about 140, about 140 to about 600, about 140 to about 580, about 140 to about 560, about 140 to about 540, about 140 to about 520, about 140 to about 500, about 140 to about 480, about 140 to about 460, about 140 to about 440, about 140 to about 420, about 140 to about 400, about 140 to about 380, about 140 to about 360, about 140 to about 340, about 140 to about 320, about 140 to about 300, about 140 to about 280, about 140 to about 260, about 140 to about 240, about 140 to about 220, about 140 to about 200, about 140 to about 190, about 140 to about 180, about 140 to about 170, about 140 to about 160, about 140 to about 150, about 150 to about 600, about 150 to about 580, about 150 to about 560, about 150 to about 540, about 150 to about 520, about 150 to about 500, about 150 to about 480, about 150 to about 460, about 150 to about 440, about 150 to about 420, about 150 to about 400, about 150 to about 380, about 150 to about 360, about 150 to about 340, about 150 to about 320, about 150 to about 300, about 150 to about 280, about 150 to about 260, about 150 to about 240, about 150 to about 220, about 150 to about 200, about 150 to about 190, about 150 to about 180, about 150 to about 170, about 150 to about 160, about 160 to about 600, about 160 to about 580, about 160 to about 560, about 160 to about 540, about 160 to about 520, about 160 to about 500, about 160 to about 480, about 160 to about 460, about 160 to about 440, about 160 to about 420, about 160 to about 400, about 160 to about 380, about 160 to about 360, about 160 to about 340, about 160 to about 320, about 160 to about 300, about 160 to about 280, about 160 to about 260, about 160 to about 240, about 160 to about 220, about 160 to about 200, about 160 to about 190, about 160 to about 180, about 160 to about 170, about 170 to about 600, about 170 to about 580, about 170 to about 560, about 170 to about 540, about 170 to about 520, about 170 to about 500, about 170 to about 480, about 170 to about 460, about 170 to about 440, about 170 to about 420, about 170 to about 400, about 170 to about 380, about 170 to about 360, about 170 to about 340, about 170 to about 320, about 170 to about 300, about 170 to about 280, about 170 to about 260, about 170 to about 240, about 170 to about 220, about 170 to about 200, about 170 to about 190, about 170 to about 180, about 180 to about 600, about 180 to about 580, about 180 to about 560, about 180 to about 540, about 180 to about 520, about 180 to about 500, about 180 to about 480, about 180 to about 460, about 180 to about 440, about 180 to about 420, about 180 to about 400, about 180 to about 380, about 180 to about 360, about 180 to about 340, about 180 to about 320, about 180 to about 300, about 180 to about 280, about 180 to about 260, about 180 to about 240, about 180 to about 220, about 180 to about 200, about 180 to about 190, about 190 to about 600, about 190 to about 580, about 190 to about 560, about 190 to about 540, about 190 to about 520, about 190 to about 500, about 190 to about 480, about 190 to about 460, about 190 to about 440, about 190 to about 420, about 190 to about 400, about 190 to about 380, about 190 to about 360, about 190 to about 340, about 190 to about 320, about 190 to about 300, about 190 to about 280, about 190 to about 260, about 190 to about 240, about 190 to about 220, about 190 to about 200, about 200 to about 600, about 200 to about 580, about 200 to about 560, about 200 to about 540, about 200 to about 520, about 200 to about 500, about 200 to about 480, about 200 to about 460, about 200 to about 440, about 200 to about 420, about 200 to about 400, about 200 to about 380, about 200 to about 360, about 200 to about 340, about 200 to about 320, about 200 to about 300, about 200 to about 280, about 200 to about 260, about 200 to about 240, about 200 to about 220, about 220 to about 600, about 220 to about 580, about 220 to about 560, about 220 to about 540, about 220 to about 520, about 220 to about 500, about 220 to about 480, about 220 to about 460, about 220 to about 440, about 220 to about 420, about 220 to about 400, about 220 to about 380, about 220 to about 360, about 220 to about 340, about 220 to about 320, about 220 to about 300, about 220 to about 280, about 220 to about 260, about 220 to about 240, about 240 to about 600, about 240 to about 580, about 240 to about 560, about 240 to about 540, about 240 to about 520, about 240 to about 500, about 240 to about 480, about 240 to about 460, about 240 to about 440, about 240 to about 420, about 240 to about 400, about 240 to about 380, about 240 to about 360, about 240 to about 340, about 240 to about 320, about 240 to about 300, about 240 to about 280, about 240 to about 260, about 260 to about 600, about 260 to about 580, about 260 to about 560, about 260 to about 540, about 260 to about 520, about 260 to about 500, about 260 to about 480, about 260 to about 460, about 260 to about 440, about 260 to about 420, about 260 to about 400, about 260 to about 380, about 260 to about 360, about 260 to about 340, about 260 to about 320, about 260 to about 300, about 260 to about 280, about 280 to about 600, about 280 to about 580, about 280 to about 560, about 280 to about 540, about 280 to about 520, about 280 to about 500, about 280 to about 480, about 280 to about 460, about 280 to about 440, about 280 to about 420, about 280 to about 400, about 280 to about 380, about 280 to about 360, about 280 to about 340, about 280 to about 320, about 280 to about 300, about 300 to about 600, about 300 to about 580, about 300 to about 560, about 300 to about 540, about 300 to about 520, about 300 to about 500, about 300 to about 480, about 300 to about 460, about 300 to about 440, about 300 to about 420, about 300 to about 400, about 300 to about 380, about 300 to about 360, about 300 to about 340, about 300 to about 320, about 320 to about 600, about 320 to about 580, about 320 to about 560, about 320 to about 540, about 320 to about 520, about 320 to about 500, about 320 to about 480, about 320 to about 460, about 320 to about 440, about 320 to about 420, about 320 to about 400, about 320 to about 380, about 320 to about 360, about 320 to about 340, about 340 to about 600, about 340 to about 580, about 340 to about 560, about 340 to about 540, about 340 to about 520, about 340 to about 500, about 340 to about 480, about 340 to about 460, about 340 to about 440, about 340 to about 420, about 340 to about 400, about 340 to about 380, about 340 to about 360, about 360 to about 600, about 360 to about 580, about 360 to about 560, about 360 to about 540, about 360 to about 520, about 360 to about 500, about 360 to about 480, about 360 to about 460, about 360 to about 440, about 360 to about 420, about 360 to about 400, about 360 to about 380, about 380 to about 600, about 380 to about 580, about 380 to about 560, about 380 to about 540, about 380 to about 520, about 380 to about 500, about 380 to about 480, about 380 to about 460, about 380 to about 440, about 380 to about 420, about 380 to about 400, about 400 to about 600, about 400 to about 580, about 400 to about 560, about 400 to about 540, about 400 to about 520, about 400 to about 500, about 400 to about 480, about 400 to about 460, about 400 to about 440, about 400 to about 420, about 420 to about 600, about 420 to about 580, about 420 to about 560, about 420 to about 540, about 420 to about 520, about 420 to about 500, about 420 to about 480, about 420 to about 460, about 420 to about 440, about 440 to about 600, about 440 to about 580, about 440 to about 560, about 440 to about 540, about 440 to about 520, about 440 to about 500, about 440 to about 480, about 440 to about 460, about 460 to about 600, about 460 to about 580, about 460 to about 560, about 460 to about 540, about 460 to about 520, about 460 to about 500, about 460 to about 480, about 480 to about 600, about 480 to about 580, about 480 to about 560, about 480 to about 540, about 480 to about 520, about 480 to about 500, about 500 to about 600, about 500 to about 580, about 500 to about 560, about 500 to about 540, about 500 to about 520, about 520 to about 600, about 520 to about 580, about 520 to about 560, about 520 to about 540, about 540 to about 600, about 540 to about 580, about 540 to about 560, about 560 to about 600, about 560 to about 580, or about 580 to about 600.

[1634] Additional examples of a ratio of GI tissue concentration of the immune modulator to the blood, serum, or plasma concentration of the immune modulator include to 1.1 to 600, 1.2 to 600, 1.3 to 600, 1.4 to 600, 1.5 to 600, 1.6 to 600, 1.7 to 600, 1.8 to 600, or 1.9 to 600, such as 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 1.9.

[1635] In some examples, administration of an immune modulator using any of the devices or compositions described herein can result in a ratio of GI tissue concentration of the immune modulator to the blood, serum, or plasma concentration of the immune modulator of, e.g., about 2.8 to about 6.0, about 2.8 to about 5.8, about 2.8 to about 5.6, about 2.8 to about 5.4, about 2.8 to about 5.2, about 2.8 to about 5.0, about 2.8 to about 4.8, about 2.8 to about 4.6, about 2.8 to about 4.4, about 2.8 to about 4.2, about 2.8 to about 4.0, about 2.8 to about 3.8, about 2.8 to about 3.6, about 2.8 to about 3.4, about 2.8 to about 3.2, about 2.8 to about 3.0, about 3.0 to about 6.0, about 3.0 to about 5.8, about 3.0 to about 5.6, about 3.0 to about 5.4, about 3.0 to about 5.2, about 3.0 to about 5.0, about 3.0 to about 4.8, about 3.0 to about 4.6, about 3.0 to about 4.4, about 3.0 to about 4.2, about 3.0 to about 4.0, about 3.0 to about 3.8, about 3.0 to about 3.6, about 3.0 to about 3.4, about 3.0 to about 3.2, about 3.2 to about 6.0, about 3.2 to about 5.8, about 3.2 to about 5.6, about 3.2 to about 5.4, about 3.2 to about 5.2, about 3.2 to about 5.0, about 3.2 to about 4.8, about 3.2 to about 4.6, about 3.2 to about 4.4, about 3.2 to about 4.2, about 3.2 to about 4.0, about 3.2 to about 3.8, about 3.2 to about 3.6, about 3.2 to about 3.4, about 3.4 to about 6.0, about 3.4 to about 5.8, about 3.4 to about 5.6, about 3.4 to about 5.4, about 3.4 to about 5.2, about 3.4 to about 5.0, about 3.4 to about 4.8, about 3.4 to about 4.6, about 3.4 to about 4.4, about 3.4 to about 4.2, about 3.4 to about 4.0, about 3.4 to about 3.8, about 3.4 to about 3.6, about 3.6 to about 6.0, about 3.6 to about 5.8, about 3.6 to about 5.6, about 3.6 to about 5.4, about 3.6 to about 5.2, about 3.6 to about 5.0, about 3.6 to about 4.8, about 3.6 to about 4.6, about 3.6 to about 4.4, about 3.6 to about 4.2, about 3.6 to about 4.0, about 3.6 to about 3.8, about 3.8 to about 6.0, about 3.8 to about 5.8, about 3.8 to about 5.6, about 3.8 to about 5.4, about 3.8 to about 5.2, about 3.8 to about 5.0, about 3.8 to about 4.8, about 3.8 to about 4.6, about 3.8 to about 4.4, about 3.8 to about 4.2, about 3.8 to about 4.0, about 4.0 to about 6.0, about 4.0 to about 5.8, about 4.0 to about 5.6, about 4.0 to about 5.4, about 4.0 to about 5.2, about 4.0 to about 5.0, about 4.0 to about 4.8, about 4.0 to about 4.6, about 4.0 to about 4.4, about 4.0 to about 4.2, about 4.2 to about 6.0, about 4.2 to about 5.8, about 4.2 to about 5.6, about 4.2 to about 5.4, about 4.2 to about 5.2, about 4.2 to about 5.0, about 4.2 to about 4.8, about 4.2 to about 4.6, about 4.2 to about 4.4, about 4.4 to about 6.0, about 4.4 to about 5.8, about 4.4 to about 5.6, about 4.4 to about 5.4, about 4.4 to about 5.2, about 4.4 to about 5.0, about 4.4 to about 4.8, about 4.4 to about 4.6, about 4.6 to about 6.0, about 4.6 to about 5.8, about 4.6 to about 5.6, about 4.6 to about 5.4, about 4.6 to about 5.2, about 4.6 to about 5.0, about 4.6 to about 4.8, about 4.8 to about 6.0, about 4.8 to about 5.8, about 4.8 to about 5.6, about 4.8 to about 5.4, about 4.8 to about 5.2, about 4.8 to about 5.0, about 5.0 to about 6.0, about 5.0 to about 5.8, about 5.0 to about 5.6, about 5.0 to about 5.4, about 5.0 to about 5.2, about 5.2 to about 6.0, about 5.2 to about 5.8, about 5.2 to about 5.6, about 5.2 to about 5.4, about 5.4 to about 6.0, about 5.4 to about 5.8, about 5.4 to about 5.6, about 5.6 to about 6.0, about 5.6 to about 5.8, or about 5.8 to about 6.0. Accordingly, in some embodiments, a method of treatment disclosed herein can include determining the ratio of the level of the immune modulator in the GI tissue to the level of the immune modulator inhibitor in the blood, serum, or plasma of a subject at substantially the same time point following administration of the device is about 2.8 to about 6.0. Exemplary methods for measuring the concentration of an immune modulator in the plasma or the GI tissue of a subject are described herein. Additional methods for measuring the concentration of an immune modulator in the plasma or the GI tissue of a subject are known in the art.

[1636] Accordingly, in some embodiments, a method of treatment disclosed herein includes determining the level of the immune modulator in the GI tissue (e.g., one or more of any of the exemplary GI tissues described herein). In some embodiments, a method of treatment disclosed herein can include determining the level of immune modulator in one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa.

[1637] In some embodiments, a method of treatment disclosed herein includes determining that the level of the immune modulator in the GI tissue (e.g., one or more of any of the exemplary types of GI tissue described herein) at a time point following administration of the device is higher than the level of the immune modulator in the GI tissue at substantially the same time point following systemic administration of an equal amount of the immune modulator. In some embodiments, a method of treatment disclosed herein can include determining that the level of the immune modulator in one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa at a time point following administration of the device is higher than the level of the immune modulator in one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa at substantially the same time point following systemic administration of an equal amount of the immune modulator.

[1638] In some embodiments, a method of treatment disclosed herein includes determining the level of immune modulator in the feces of the subject. In some embodiments, a method of treatment disclosed herein includes determining the level of immune modulator in the GI tissue, e.g., in one or more (e.g., two, three, or four) of the lumen/superficial mucosa, the lamina propria, the submucosa, and the tunica muscularis/serosa within a time period of about 10 minutes to about 10 hours following administration of the device.

[1639] In some embodiments, a method of treatment as disclosed herein comprises determining the level of the immune modulator at the location of release following administration of the device.

[1640] In some embodiments, a method of treatment as disclosed herein comprises determining that the level of immune modulator at the location of release at the time point following administration of the device is higher than the level of the immune modulator at the same location of release at substantially the same time point following systemic administration of an equal amount of the immune modulator.

[1641] In some embodiments, a method of treatment as disclosed herein comprises determining the level of the immune modulator in the tissue of the subject within a time period of about 10 minutes to 10 hours following administration of the device.

[1642] Some examples of any of the methods described herein can, e.g., result in a selective suppression of a local inflammatory response (e.g., suppression in the local lymphatic system, for example, in the mesenteric lymph nodes), while maintaining the systemic immune response (e.g., blood).

[1643] FAs used herein, GI content refers to the content of the gastrointestinal (GI) tract, such as the content of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum, more particularly of the proximal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon, or of the distal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon.

[1644] In some examples, the methods described herein can result in a 1% increase to 500% increase (e.g., a 1% increase to 450% increase, a 1% increase to 400% increase, a 1% increase to 350% increase, a 1% increase to 300% increase, a 1% increase to 250% increase, a 1% increase to 200% increase, a 1% increase to 190% increase, a 1% increase to 180% increase, a 1% increase to 170% increase, a 1% increase to 160% increase, a 1% increase to 150% increase, a 1% increase to 140% increase, a 1% increase to 130% increase, a 1% increase to 120% increase, a 1% increase to 110% increase, a 1% increase to 100% increase, a 1% increase to 90% increase, a 1% increase to 80% increase, a 1% increase to 70% increase, a 1% increase to 60% increase, a 1% increase to 50% increase, a 1% increase to 40% increase, a 1% increase to 30% increase, a 1% increase to 25% increase, a 1% increase to 20% increase, a 1% increase to 15% increase, a 1% increase to 10% increase, a 1% increase to 5% increase, a 5% increase to 500% increase, a 5% increase to 450% increase, a 5% increase to 400% increase, a 5% increase to 350% increase, a 5% increase to 300% increase, a 5% increase to 250% increase, a 5% increase to 200% increase, a 5% increase to 190% increase, a 5% increase to 180% increase, a 5% increase to 170% increase, a 5% increase to 160% increase, a 5% increase to 150% increase, a 5% increase to 140% increase, a 5% increase to 130% increase, a 5% increase to 120% increase, a 5% increase to 110% increase, a 5% increase to 100% increase, a 5% increase to 90% increase, a 5% increase to 80% increase, a 5% increase to 70% increase, a 5% increase to 60% increase, a 5% increase to 50% increase, a 5% increase to 40% increase, a 5% increase to 30% increase, a 5% increase to 25% increase, a 5% increase to 20% increase, a 5% increase to 15% increase, a 5% increase to 10% increase, a 10% increase to 500% increase, a 10% increase to 450% increase, a 10% increase to 400% increase, a 10% increase to 350% increase, a 10% increase to 300% increase, a 10% increase to 250% increase, a 10% increase to 200% increase, a 10% increase to 190% increase, a 10% increase to 180% increase, a 10% increase to 170% increase, a 10% increase to 160% increase, a 10% increase to 150% increase, a 10% increase to 140% increase, a 10% increase to 130% increase, a 10% increase to 120% increase, a 10% increase to 110% increase, a 10% increase to 100% increase, a 10% increase to 90% increase, a 10% increase to 80% increase, a 10% increase to 70% increase, a 10% increase to 60% increase, a 10% increase to 50% increase, a 10% increase to 40% increase, a 10% increase to 30% increase, a 10% increase to 25% increase, a 10% increase to 20% increase, a 10% increase to 15% increase, a 15% increase to 500% increase, a 15% increase to 450% increase, a 15% increase to 400% increase, a 15% increase to 350% increase, a 15% increase to 300% increase, a 15% increase to 250% increase, a 15% increase to 200% increase, a 15% increase to 190% increase, a 15% increase to 180% increase, a 15% increase to 170% increase, a 15% increase to 160% increase, a 15% increase to 150% increase, a 15% increase to 140% increase, a 15% increase to 130% increase, a 15% increase to 120% increase, a 15% increase to 110% increase, a 15% increase to 100% increase, a 15% increase to 90% increase, a 15% increase to 80% increase, a 15% increase to 70% increase, a 15% increase to 60% increase, a 15% increase to 50% increase, a 15% increase to 40% increase, a 15% increase to 30% increase, a 15% increase to 25% increase, a 15% increase to 20% increase, a 20% increase to 500% increase, a 20% increase to 450% increase, a 20% increase to 400% increase, a 20% increase to 350% increase, a 20% increase to 300% increase, a 20% increase to 250% increase, a 20% increase to 200% increase, a 20% increase to 190% increase, a 20% increase to 180% increase, a 20% increase to 170% increase, a 20% increase to 160% increase, a 20% increase to 150% increase, a 20% increase to 140% increase, a 20% increase to 130% increase, a 20% increase to 120% increase, a 20% increase to 110% increase, a 20% increase to 100% increase, a 20% increase to 90% increase, a 20% increase to 80% increase, a 20% increase to 70% increase, a 20% increase to 60% increase, a 20% increase to 50% increase, a 20% increase to 40% increase, a 20% increase to 30% increase, a 20% increase to 25% increase, a 25% increase to 500% increase, a 25% increase to 450% increase, a 25% increase to 400% increase, a 25% increase to 350% increase, a 25% increase to 300% increase, a 25% increase to 250% increase, a 25% increase to 200% increase, a 25% increase to 190% increase, a 25% increase to 180% increase, a 25% increase to 170% increase, a 25% increase to 160% increase, a 25% increase to 150% increase, a 25% increase to 140% increase, a 25% increase to 130% increase, a 25% increase to 120% increase, a 25% increase to 110% increase, a 25% increase to 100% increase, a 25% increase to 90% increase, a 25% increase to 80% increase, a 25% increase to 70% increase, a 25% increase to 60% increase, a 25% increase to 50% increase, a 25% increase to 40% increase, a 25% increase to 30% increase, a 30% increase to 500% increase, a 30% increase to 450% increase, a 30% increase to 400% increase, a 30% increase to 350% increase, a 30% increase to 300% increase, a 30% increase to 250% increase, a 30% increase to 200% increase, a 30% increase to 190% increase, a 30% increase to 180% increase, a 30% increase to 170% increase, a 30% increase to 160% increase, a 30% increase to 150% increase, a 30% increase to 140% increase, a 30% increase to 130% increase, a 30% increase to 120% increase, a 30% increase to 110% increase, a 30% increase to 100% increase, a 30% increase to 90% increase, a 30% increase to 80% increase, a 30% increase to 70% increase, a 30% increase to 60% increase, a 30% increase to 50% increase, a 30% increase to 40% increase, a 40% increase to 500% increase, a 40% increase to 450% increase, a 40% increase to 400% increase, a 40% increase to 350% increase, a 40% increase to 300% increase, a 40% increase to 250% increase, a 40% increase to 200% increase, a 40% increase to 190% increase, a 40% increase to 180% increase, a 40% increase to 170% increase, a 40% increase to 160% increase, a 40% increase to 150% increase, a 40% increase to 140% increase, a 40% increase to 130% increase, a 40% increase to 120% increase, a 40% increase to 110% increase, a 40% increase to 100% increase, a 40% increase to 90% increase, a 40% increase to 80% increase, a 40% increase to 70% increase, a 40% increase to 60% increase, a 40% increase to 50% increase, a 50% increase to 500% increase, a 50% increase to 450% increase, a 50% increase to 400% increase, a 50% increase to 350% increase, a 50% increase to 300% increase, a 50% increase to 250% increase, a 50% increase to 200% increase, a 50% increase to 190% increase, a 50% increase to 180% increase, a 50% increase to 170% increase, a 50% increase to 160% increase, a 50% increase to 150% increase, a 50% increase to 140% increase, a 50% increase to 130% increase, a 50% increase to 120% increase, a 50% increase to 110% increase, a 50% increase to 100% increase, a 50% increase to 90% increase, a 50% increase to 80% increase, a 50% increase to 70% increase, a 50% increase to 60% increase, a 60% increase to 500% increase, a 60% increase to 450% increase, a 60% increase to 400% increase, a 60% increase to 350% increase, a 60% increase to 300% increase, a 60% increase to 250% increase, a 60% increase to 200% increase, a 60% increase to 190% increase, a 60% increase to 180% increase, a 60% increase to 170% increase, a 60% increase to 160% increase, a 60% increase to 150% increase, a 60% increase to 140% increase, a 60% increase to 130% increase, a 60% increase to 120% increase, a 60% increase to 110% increase, a 60% increase to 100% increase, a 60% increase to 90% increase, a 60% increase to 80% increase, a 60% increase to 70% increase, a 70% increase to 500% increase, a 70% increase to 450% increase, a 70% increase to 400% increase, a 70% increase to 350% increase, a 70% increase to 300% increase, a 70% increase to 250% increase, a 70% increase to 200% increase, a 70% increase to 190% increase, a 70% increase to 180% increase, a 70% increase to 170% increase, a 70% increase to 160% increase, a 70% increase to 150% increase, a 70% increase to 140% increase, a 70% increase to 130% increase, a 70% increase to 120% increase, a 70% increase to 110% increase, a 70% increase to 100% increase, a 70% increase to 90% increase, a 70% increase to 80% increase, a 80% increase to 500% increase, a 80% increase to 450% increase, a 80% increase to 400% increase, a 80% increase to 350% increase, a 80% increase to 300% increase, a 80% increase to 250% increase, a 80% increase to 200% increase, a 80% increase to 190% increase, a 80% increase to 180% increase, a 80% increase to 170% increase, a 80% increase to 160% increase, a 80% increase to 150% increase, a 80% increase to 140% increase, a 80% increase to 130% increase, a 80% increase to 120% increase, a 80% increase to 110% increase, a 80% increase to 100% increase, a 80% increase to 90% increase, a 90% increase to 500% increase, a 90% increase to 450% increase, a 90% increase to 400% increase, a 90% increase to 350% increase, a 90% increase to 300% increase, a 90% increase to 250% increase, a 90% increase to 200% increase, a 90% increase to 190% increase, a 90% increase to 180% increase, a 90% increase to 170% increase, a 90% increase to 160% increase, a 90% increase to 150% increase, a 90% increase to 140% increase, a 90% increase to 130% increase, a 90% increase to 120% increase, a 90% increase to 110% increase, a 90% increase to 100% increase, a 100% increase to 500% increase, a 100% increase to 450% increase, a 100% increase to 400% increase, a 100% increase to 350% increase, a 100% increase to 300% increase, a 100% increase to 250% increase, a 100% increase to 200% increase, a 100% increase to 190% increase, a 100% increase to 180% increase, a 100% increase to 170% increase, a 100% increase to 160% increase, a 100% increase to 150% increase, a 100% increase to 140% increase, a 100% increase to 130% increase, a 100% increase to 120% increase, a 100% increase to 110% increase, a 110% increase to 500% increase, a 110% increase to 450% increase, a 110% increase to 400% increase, a 110% increase to 350% increase, a 110% increase to 300% increase, a 110% increase to 250% increase, a 110% increase to 200% increase, a 110% increase to 190% increase, a 110% increase to 180% increase, a 110% increase to 170% increase, a 110% increase to 160% increase, a 110% increase to 150% increase, a 110% increase to 140% increase, a 110% increase to 130% increase, a 110% increase to 120% increase, a 120% increase to 500% increase, a 120% increase to 450% increase, a 120% increase to 400% increase, a 120% increase to 350% increase, a 120% increase to 300% increase, a 120% increase to 250% increase, a 120% increase to 200% increase, a 120% increase to 190% increase, a 120% increase to 180% increase, a 120% increase to 170% increase, a 120% increase to 160% increase, a 120% increase to 150% increase, a 120% increase to 140% increase, a 120% increase to 130% increase, a 130% increase to 500% increase, a 130% increase to 450% increase, a 130% increase to 400% increase, a 130% increase to 350% increase, a 130% increase to 300% increase, a 130% increase to 250% increase, a 130% increase to 200% increase, a 130% increase to 190% increase, a 130% increase to 180% increase, a 130% increase to 170% increase, a 130% increase to 160% increase, a 130% increase to 150% increase, a 130% increase to 140% increase, a 140% increase to 500% increase, a 140% increase to 450% increase, a 140% increase to 400% increase, a 140% increase to 350% increase, a 140% increase to 300% increase, a 140% increase to 250% increase, a 140% increase to 200% increase, a 140% increase to 190% increase, a 140% increase to 180% increase, a 140% increase to 170% increase, a 140% increase to 160% increase, a 140% increase to 150% increase, a 150% increase to 500% increase, a 150% increase to 450% increase, a 150% increase to 400% increase, a 150% increase to 350% increase, a 150% increase to 300% increase, a 150% increase to 250% increase, a 150% increase to 200% increase, a 150% increase to 190% increase, a 150% increase to 180% increase, a 150% increase to 170% increase, a 150% increase to 160% increase, a 160% increase to 500% increase, a 160% increase to 450% increase, a 160% increase to 400% increase, a 160% increase to 350% increase, a 160% increase to 300% increase, a 160% increase to 250% increase, a 160% increase to 200% increase, a 160% increase to 190% increase, a 160% increase to 180% increase, a 160% increase to 170% increase, a 170% increase to 500% increase, a 170% increase to 450% increase, a 170% increase to 400% increase, a 170% increase to 350% increase, a 170% increase to 300% increase, a 170% increase to 250% increase, a 170% increase to 200% increase, a 170% increase to 190% increase, a 170% increase to 180% increase, a 180% increase to 500% increase, a 180% increase to 450% increase, a 180% increase to 400% increase, a 180% increase to 350% increase, a 180% increase to 300% increase, a 180% increase to 250% increase, a 180% increase to 200% increase, a 180% increase to 190% increase, a 190% increase to 500% increase, a 190% increase to 450% increase, a 190% increase to 400% increase, a 190% increase to 350% increase, a 190% increase to 300% increase, a 190% increase to 250% increase, a 190% increase to 200% increase, a 200% increase to 500% increase, a 200% increase to 450% increase, a 200% increase to 400% increase, a 200% increase to 350% increase, a 200% increase to 300% increase, a 200% increase to 250% increase, a 250% increase to 500% increase, a 250% increase to 450% increase, a 250% increase to 400% increase, a 250% increase to 350% increase, a 250% increase to 300% increase, a 300% increase to 500% increase, a 300% increase to 450% increase, a 300% increase to 400% increase, a 300% increase to 350% increase, a 350% increase to 500% increase, a 350% increase to 450% increase, a 350% increase to 400% increase, a 400% increase to 500% increase, a 400% increase to 450% increase, or a 450% increase to 500% increase) in one or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) of: the plasma, serum, or blood level of IL-6; the plasma, serum, or blood level of IL-2; the plasma, serum, or blood level of IL-I; the plasma, serum, or blood level of TNF; the plasma, serum, or blood level of IL-17A; the plasma, serum, or blood level of IL-22; the plasma, serum, or blood level of interferon-; the level of blood Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells); the level of 47 expression in blood cells, and the level of 47 expression in Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells) in mesenteric lymph nodes, e.g., each as compared to the corresponding level in a subject systemically administered the same dose of the same immune modulator. Methods for determining the plasma, serum, or blood level of IL-6; the plasma, serum, or blood level of IL-2; the plasma, serum, or blood level of IL-1; the plasma, serum, or blood level of TNF; the plasma, serum, or blood level of IL-17A; the plasma, serum, or blood level of IL-22; the plasma, serum, or blood level of interferon-; the level of blood Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells); and the level of 47 expression in blood cells; and the level of 47 expression in blood cells are known in the art.

[1645] In some examples, the methods described herein can result in a 1% decrease to 99% decrease (e.g., a 1% decrease to 95% decrease, a 1% decrease to 90% decrease, a 1% decrease to 85% decrease, a 1% decrease to 80% decrease, a 1% decrease to 75% decrease, a 1% decrease to 70% decrease, a 1% decrease to 65% decrease, a 1% decrease to 60% decrease, a 1% decrease to 55% decrease, a 1% decrease to 50% decrease, a 1% decrease to 45% decrease, a 1% decrease to 40% decrease, a 1% decrease to 35% decrease, a 1% decrease to 30% decrease, a 1% decrease to 25% decrease, a 1% decrease to 20% decrease, a 1% decrease to 15% decrease, a 1% decrease to 10% decrease, a 1% decrease to 5% decrease, a 5% decrease to 99% decrease, a 5% decrease to 95% decrease, a 5% decrease to 90% decrease, a 5% decrease to 85% decrease, a 5% decrease to 80% decrease, a 5% decrease to 75% decrease, a 5% decrease to 70% decrease, a 5% decrease to 65% decrease, a 5% decrease to 60% decrease, a 5% decrease to 55% decrease, a 5% decrease to 50% decrease, a 5% decrease to 45% decrease, a 5% decrease to 40% decrease, a 5% decrease to 35% decrease, a 5% decrease to 30% decrease, a 5% decrease to 25% decrease, a 5% decrease to 20% decrease, a 5% decrease to 15% decrease, a 5% decrease to 10% decrease, a 10% decrease to 99% decrease, a 10% decrease to 95% decrease, a 10% decrease to 90% decrease, a 10% decrease to 85% decrease, a 10% decrease to 80% decrease, a 10% decrease to 75% decrease, a 10% decrease to 70% decrease, a 10% decrease to 65% decrease, a 10% decrease to 60% decrease, a 10% decrease to 55% decrease, a 10% decrease to 50% decrease, a 10% decrease to 45% decrease, a 10% decrease to 40% decrease, a 10% decrease to 35% decrease, a 10% decrease to 30% decrease, a 10% decrease to 25% decrease, a 10% decrease to 20% decrease, a 10% decrease to 15% decrease, a 15% decrease to 99% decrease, a 15% decrease to 95% decrease, a 15% decrease to 90% decrease, a 15% decrease to 85% decrease, a 15% decrease to 80% decrease, a 15% decrease to 75% decrease, a 15% decrease to 70% decrease, a 15% decrease to 65% decrease, a 15% decrease to 60% decrease, a 15% decrease to 55% decrease, a 15% decrease to 50% decrease, a 15% decrease to 45% decrease, a 15% decrease to 40% decrease, a 15% decrease to 35% decrease, a 15% decrease to 30% decrease, a 15% decrease to 25% decrease, a 15% decrease to 20% decrease, a 20% decrease to 99% decrease, a 20% decrease to 95% decrease, a 20% decrease to 90% decrease, a 20% decrease to 85% decrease, a 20% decrease to 80% decrease, a 20% decrease to 75% decrease, a 20% decrease to 70% decrease, a 20% decrease to 65% decrease, a 20% decrease to 60% decrease, a 20% decrease to 55% decrease, a 20% decrease to 50% decrease, a 20% decrease to 45% decrease, a 20% decrease to 40% decrease, a 20% decrease to 35% decrease, a 20% decrease to 30% decrease, a 20% decrease to 25% decrease, a 25% decrease to 99% decrease, a 25% decrease to 95% decrease, a 25% decrease to 90% decrease, a 25% decrease to 85% decrease, a 25% decrease to 80% decrease, a 25% decrease to 75% decrease, a 25% decrease to 70% decrease, a 25% decrease to 65% decrease, a 25% decrease to 60% decrease, a 25% decrease to 55% decrease, a 25% decrease to 50% decrease, a 25% decrease to 45% decrease, a 25% decrease to 40% decrease, a 25% decrease to 35% decrease, a 25% decrease to 30% decrease, a 30% decrease to 99% decrease, a 30% decrease to 95% decrease, a 30% decrease to 90% decrease, a 30% decrease to 85% decrease, a 30% decrease to 80% decrease, a 30% decrease to 75% decrease, a 30% decrease to 70% decrease, a 30% decrease to 65% decrease, a 30% decrease to 60% decrease, a 30% decrease to 55% decrease, a 30% decrease to 50% decrease, a 30% decrease to 45% decrease, a 30% decrease to 40% decrease, a 30% decrease to 35% decrease, a 35% decrease to 99% decrease, a 35% decrease to 95% decrease, a 35% decrease to 90% decrease, a 35% decrease to 85% decrease, a 35% decrease to 80% decrease, a 35% decrease to 75% decrease, a 35% decrease to 70% decrease, a 35% decrease to 65% decrease, a 35% decrease to 60% decrease, a 35% decrease to 55% decrease, a 35% decrease to 50% decrease, a 35% decrease to 45% decrease, a 35% decrease to 40% decrease, a 40% decrease to 99% decrease, a 40% decrease to 95% decrease, a 40% decrease to 90% decrease, a 40% decrease to 85% decrease, a 40% decrease to 80% decrease, a 40% decrease to 75% decrease, a 40% decrease to 70% decrease, a 40% decrease to 65% decrease, a 40% decrease to 60% decrease, a 40% decrease to 55% decrease, a 40% decrease to 50% decrease, a 40% decrease to 45% decrease, a 45% decrease to 99% decrease, a 45% decrease to 95% decrease, a 45% decrease to 90% decrease, a 45% decrease to 85% decrease, a 45% decrease to 80% decrease, a 45% decrease to 75% decrease, a 45% decrease to 70% decrease, a 45% decrease to 65% decrease, a 45% decrease to 60% decrease, a 45% decrease to 55% decrease, a 45% decrease to 50% decrease, a 50% decrease to 99% decrease, a 50% decrease to 95% decrease, a 50% decrease to 90% decrease, a 50% decrease to 85% decrease, a 50% decrease to 80% decrease, a 50% decrease to 75% decrease, a 50% decrease to 70% decrease, a 50% decrease to 65% decrease, a 50% decrease to 60% decrease, a 50% decrease to 55% decrease, a 55% decrease to 99% decrease, a 55% decrease to 95% decrease, a 55% decrease to 90% decrease, a 55% decrease to 85% decrease, a 55% decrease to 80% decrease, a 55% decrease to 75% decrease, a 55% decrease to 70% decrease, a 55% decrease to 65% decrease, a 55% decrease to 60% decrease, a 60% decrease to 99% decrease, a 60% decrease to 95% decrease, a 60% decrease to 90% decrease, a 60% decrease to 85% decrease, a 60% decrease to 80% decrease, a 60% decrease to 75% decrease, a 60% decrease to 70% decrease, a 60% decrease to 65% decrease, a 65% decrease to 99% decrease, a 65% decrease to 95% decrease, a 65% decrease to 90% decrease, a 65% decrease to 85% decrease, a 65% decrease to 80% decrease, a 65% decrease to 75% decrease, a 65% decrease to 70% decrease, a 70% decrease to 99% decrease, a 70% decrease to 95% decrease, a 70% decrease to 90% decrease, a 70% decrease to 85% decrease, a 70% decrease to 80% decrease, a 70% decrease to 75% decrease, a 75% decrease to 99% decrease, a 75% decrease to 95% decrease, a 75% decrease to 90% decrease, a 75% decrease to 85% decrease, a 75% decrease to 80% decrease, a 80% decrease to 99% decrease, a 80% decrease to 95% decrease, a 80% decrease to 90% decrease, a 80% decrease to 85% decrease, a 85% decrease to 99% decrease, a 85% decrease to 95% decrease, a 85% decrease to 90% decrease, a 90% decrease to 99% decrease, a 90% decrease to 95% decrease, or a 95% decrease to 99% decrease) in the the level of Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells) in mesenteric lymph nodes and/or the level of Th memory cells in Peyer's patches, e.g., as compared to the corresponding level in a subject systemically administered the same dose of the immune modulator. Methods for determining the level of Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells) in Peyer's patches, and the level of Th memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells) in mesenteric lymph nodes are known in the art.

[1646] In some embodiments, the immune modulator is delivered to the location by a process that does not comprise systemic transport of the immune modulator.

[1647] In some embodiments, the amount of the immune modulator that is administered is from about 1 mg to about 650 mg. In some embodiments, the amount of immune modulator that is administered is from about 1 mg to about 600 mg. In some embodiments, the amount of the immune modulator that is administered is from about 1 mg to about 500 mg. In some embodiments, the amount of the immune modulator that is administered is from about 1 mg to about 100 mg. In some embodiments, the amount of the immune modulator that is administered is from about 5 mg to about 40 mg. In some embodiments, the amount of the immune modulator inhibitor is administered as an escalating dose of 10 mg, followed by 20 mg, followed by 30 mg; or an escalating dose of 20 mg, followed by 30 mg, followed by 50 mg.

[1648] In some embodiments, the amount of the immune modulator inhibitor is administered in a dose of, e.g., about 1 mg to about 300 mg, about 1 mg to about 250 mg, about 1 mg to about 200 mg, about 1 mg to about 195 mg, about 1 mg to about 190 mg, about 1 mg to about 185 mg, about 1 mg to about 180 mg, about 1 mg to about 175 mg, about 1 mg to about 170 mg, about 1 mg to about 165 mg, about 1 mg to about 160 mg, about 1 mg to about 155 mg, about 1 mg to about 150 mg, about 1 mg to about 145 mg, about 1 mg to about 140 mg, about 1 mg to about 135 mg, about 1 mg to about 130 mg, about 1 mg to about 125 mg, about 1 mg to about 120 mg, about 1 mg to about 115 mg, about 1 mg to about 110 mg, about 1 mg to about 105 mg, about 1 mg to about 100 mg, about 1 mg to about 95 mg, about 1 mg to about 90 mg, about 1 mg to about 85 mg, about 1 mg to about 80 mg, about 1 mg to about 75 mg, about 1 mg to about 70 mg, about 1 mg to about 65 mg, about 1 mg to about 60 mg, about 1 mg to about 55 mg, about 1 mg to about 50 mg, about 1 mg to about 45 mg, about 1 mg to about 40 mg, about 1 mg to about 35 mg, about 1 mg to about 30 mg, about 1 mg to about 25 mg, about 1 mg to about 20 mg, about 1 mg to about 15 mg, about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 5 mg to about 200 mg, about 5 mg to about 195 mg, about 5 mg to about 190 mg, about 5 mg to about 185 mg, about 5 mg to about 180 mg, about 5 mg to about 175 mg, about 5 mg to about 170 mg, about 5 mg to about 165 mg, about 5 mg to about 160 mg, about 5 mg to about 155 mg, about 5 mg to about 150 mg, about 5 mg to about 145 mg, about 5 mg to about 140 mg, about 5 mg to about 135 mg, about 5 mg to about 130 mg, about 5 mg to about 125 mg, about 5 mg to about 120 mg, about 5 mg to about 115 mg, about 5 mg to about 110 mg, about 5 mg to about 105 mg, about 5 mg to about 100 mg, about 5 mg to about 95 mg, about 5 mg to about 90 mg, about 5 mg to about 85 mg, about 5 mg to about 80 mg, about 5 mg to about 75 mg, about 5 mg to about 70 mg, about 5 mg to about 65 mg, about 5 mg to about 60 mg, about 5 mg to about 55 mg, about 5 mg to about 50 mg, about 5 mg to about 45 mg, about 5 mg to about 40 mg, about 5 mg to about 35 mg, about 5 mg to about 30 mg, about 5 mg to about 25 mg, about 5 mg to about 20 mg, about 5 mg to about 15 mg, about 5 mg to about 10 mg, about 10 mg to about 200 mg, about 10 mg to about 195 mg, about 10 mg to about 190 mg, about 10 mg to about 185 mg, about 10 mg to about 180 mg, about 10 mg to about 175 mg, about 10 mg to about 170 mg, about 10 mg to about 165 mg, about 10 mg to about 160 mg, about 10 mg to about 155 mg, about 10 mg to about 150 mg, about 10 mg to about 145 mg, about 10 mg to about 140 mg, about 10 mg to about 135 mg, about 10 mg to about 130 mg, about 10 mg to about 125 mg, about 10 mg to about 120 mg, about 10 mg to about 115 mg, about 10 mg to about 110 mg, about 10 mg to about 105 mg, about 10 mg to about 100 mg, about 10 mg to about 95 mg, about 10 mg to about 90 mg, about 10 mg to about 85 mg, about 10 mg to about 80 mg, about 10 mg to about 75 mg, about 10 mg to about 70 mg, about 10 mg to about 65 mg, about 10 mg to about 60 mg, about 10 mg to about 55 mg, about 10 mg to about 50 mg, about 10 mg to about 45 mg, about 10 mg to about 40 mg, about 10 mg to about 35 mg, about 10 mg to about 30 mg, about 10 mg to about 25 mg, about 10 mg to about 20 mg, about 10 mg to about 15 mg, about 15 mg to about 200 mg, about 15 mg to about 195 mg, about 15 mg to about 190 mg, about 15 mg to about 185 mg, about 15 mg to about 180 mg, about 15 mg to about 175 mg, about 15 mg to about 170 mg, about 15 mg to about 165 mg, about 15 mg to about 160 mg, about 15 mg to about 155 mg, about 15 mg to about 150 mg, about 15 mg to about 145 mg, about 15 mg to about 140 mg, about 15 mg to about 135 mg, about 15 mg to about 130 mg, about 15 mg to about 125 mg, about 15 mg to about 120 mg, about 15 mg to about 115 mg, about 15 mg to about 110 mg, about 15 mg to about 105 mg, about 15 mg to about 100 mg, about 15 mg to about 95 mg, about 15 mg to about 90 mg, about 15 mg to about 85 mg, about 15 mg to about 80 mg, about 15 mg to about 75 mg, about 15 mg to about 70 mg, about 15 mg to about 65 mg, about 15 mg to about 60 mg, about 15 mg to about 55 mg, about 15 mg to about 50 mg, about 15 mg to about 45 mg, about 15 mg to about 40 mg, about 15 mg to about 35 mg, about 15 mg to about 30 mg, about 15 mg to about 25 mg, about 15 mg to about 20 mg, about 20 mg to about 200 mg, about 20 mg to about 195 mg, about 20 mg to about 190 mg, about 20 mg to about 185 mg, about 20 mg to about 180 mg, about 20 mg to about 175 mg, about 20 mg to about 170 mg, about 20 mg to about 165 mg, about 20 mg to about 160 mg, about 20 mg to about 155 mg, about 20 mg to about 150 mg, about 20 mg to about 145 mg, about 20 mg to about 140 mg, about 20 mg to about 135 mg, about 20 mg to about 130 mg, about 20 mg to about 125 mg, about 20 mg to about 120 mg, about 20 mg to about 115 mg, about 20 mg to about 110 mg, about 20 mg to about 105 mg, about 20 mg to about 100 mg, about 20 mg to about 95 mg, about 20 mg to about 90 mg, about 20 mg to about 85 mg, about 20 mg to about 80 mg, about 20 mg to about 75 mg, about 20 mg to about 70 mg, about 20 mg to about 65 mg, about 20 mg to about 60 mg, about 20 mg to about 55 mg, about 20 mg to about 50 mg, about 20 mg to about 45 mg, about 20 mg to about 40 mg, about 20 mg to about 35 mg, about 20 mg to about 30 mg, about 20 mg to about 25 mg, about 25 mg to about 200 mg, about 25 mg to about 195 mg, about 25 mg to about 190 mg, about 25 mg to about 185 mg, about 25 mg to about 180 mg, about 25 mg to about 175 mg, about 25 mg to about 170 mg, about 25 mg to about 165 mg, about 25 mg to about 160 mg, about 25 mg to about 155 mg, about 25 mg to about 150 mg, about 25 mg to about 145 mg, about 25 mg to about 140 mg, about 25 mg to about 135 mg, about 25 mg to about 130 mg, about 25 mg to about 125 mg, about 25 mg to about 120 mg, about 25 mg to about 115 mg, about 25 mg to about 110 mg, about 25 mg to about 105 mg, about 25 mg to about 100 mg, about 25 mg to about 95 mg, about 25 mg to about 90 mg, about 25 mg to about 85 mg, about 25 mg to about 80 mg, about 25 mg to about 75 mg, about 25 mg to about 70 mg, about 25 mg to about 65 mg, about 25 mg to about 60 mg, about 25 mg to about 55 mg, about 25 mg to about 50 mg, about 25 mg to about 45 mg, about 25 mg to about 40 mg, about 25 mg to about 35 mg, about 25 mg to about 30 mg, about 30 mg to about 200 mg, about 30 mg to about 195 mg, about 30 mg to about 190 mg, about 30 mg to about 185 mg, about 30 mg to about 180 mg, about 30 mg to about 175 mg, about 30 mg to about 170 mg, about 30 mg to about 165 mg, about 30 mg to about 160 mg, about 30 mg to about 155 mg, about 30 mg to about 150 mg, about 30 mg to about 145 mg, about 30 mg to about 140 mg, about 30 mg to about 135 mg, about 30 mg to about 130 mg, about 30 mg to about 125 mg, about 30 mg to about 120 mg, about 30 mg to about 115 mg, about 30 mg to about 110 mg, about 30 mg to about 105 mg, about 30 mg to about 100 mg, about 30 mg to about 95 mg, about 30 mg to about 90 mg, about 30 mg to about 85 mg, about 30 mg to about 80 mg, about 30 mg to about 75 mg, about 30 mg to about 70 mg, about 30 mg to about 65 mg, about 30 mg to about 60 mg, about 30 mg to about 55 mg, about 30 mg to about 50 mg, about 30 mg to about 45 mg, about 30 mg to about 40 mg, about 30 mg to about 35 mg, about 35 mg to about 200 mg, about 35 mg to about 195 mg, about 35 mg to about 190 mg, about 35 mg to about 185 mg, about 35 mg to about 180 mg, about 35 mg to about 175 mg, about 35 mg to about 170 mg, about 35 mg to about 165 mg, about 35 mg to about 160 mg, about 35 mg to about 155 mg, about 35 mg to about 150 mg, about 35 mg to about 145 mg, about 35 mg to about 140 mg, about 35 mg to about 135 mg, about 35 mg to about 130 mg, about 35 mg to about 125 mg, about 35 mg to about 120 mg, about 35 mg to about 115 mg, about 35 mg to about 110 mg, about 35 mg to about 105 mg, about 35 mg to about 100 mg, about 35 mg to about 95 mg, about 35 mg to about 90 mg, about 35 mg to about 85 mg, about 35 mg to about 80 mg, about 35 mg to about 75 mg, about 35 mg to about 70 mg, about 35 mg to about 65 mg, about 35 mg to about 60 mg, about 35 mg to about 55 mg, about 35 mg to about 50 mg, about 35 mg to about 45 mg, about 35 mg to about 40 mg, about 40 mg to about 200 mg, about 40 mg to about 195 mg, about 40 mg to about 190 mg, about 40 mg to about 185 mg, about 40 mg to about 180 mg, about 40 mg to about 175 mg, about 40 mg to about 170 mg, about 40 mg to about 165 mg, about 40 mg to about 160 mg, about 40 mg to about 155 mg, about 40 mg to about 150 mg, about 40 mg to about 145 mg, about 40 mg to about 140 mg, about 40 mg to about 135 mg, about 40 mg to about 130 mg, about 40 mg to about 125 mg, about 40 mg to about 120 mg, about 40 mg to about 115 mg, about 40 mg to about 110 mg, about 40 mg to about 105 mg, about 40 mg to about 100 mg, about 40 mg to about 95 mg, about 40 mg to about 90 mg, about 40 mg to about 85 mg, about 40 mg to about 80 mg, about 40 mg to about 75 mg, about 40 mg to about 70 mg, about 40 mg to about 65 mg, about 40 mg to about 60 mg, about 40 mg to about 55 mg, about 40 mg to about 50 mg, about 40 mg to about 45 mg, about 45 mg to about 200 mg, about 45 mg to about 195 mg, about 45 mg to about 190 mg, about 45 mg to about 185 mg, about 45 mg to about 180 mg, about 45 mg to about 175 mg, about 45 mg to about 170 mg, about 45 mg to about 165 mg, about 45 mg to about 160 mg, about 45 mg to about 155 mg, about 45 mg to about 150 mg, about 45 mg to about 145 mg, about 45 mg to about 140 mg, about 45 mg to about 135 mg, about 45 mg to about 130 mg, about 45 mg to about 125 mg, about 45 mg to about 120 mg, about 45 mg to about 115 mg, about 45 mg to about 110 mg, about 45 mg to about 105 mg, about 45 mg to about 100 mg, about 45 mg to about 95 mg, about 45 mg to about 90 mg, about 45 mg to about 85 mg, about 45 mg to about 80 mg, about 45 mg to about 75 mg, about 45 mg to about 70 mg, about 45 mg to about 65 mg, about 45 mg to about 60 mg, about 45 mg to about 55 mg, about 45 mg to about 50 mg, about 50 mg to about 200 mg, about 50 mg to about 195 mg, about 50 mg to about 190 mg, about 50 mg to about 185 mg, about 50 mg to about 180 mg, about 50 mg to about 175 mg, about 50 mg to about 170 mg, about 50 mg to about 165 mg, about 50 mg to about 160 mg, about 50 mg to about 155 mg, about 50 mg to about 150 mg, about 50 mg to about 145 mg, about 50 mg to about 140 mg, about 50 mg to about 135 mg, about 50 mg to about 130 mg, about 50 mg to about 125 mg, about 50 mg to about 120 mg, about 50 mg to about 115 mg, about 50 mg to about 110 mg, about 50 mg to about 105 mg, about 50 mg to about 100 mg, about 50 mg to about 95 mg, about 50 mg to about 90 mg, about 50 mg to about 85 mg, about 50 mg to about 80 mg, about 50 mg to about 75 mg, about 50 mg to about 70 mg, about 50 mg to about 65 mg, about 50 mg to about 60 mg, about 50 mg to about 55 mg, about 55 mg to about 200 mg, about 55 mg to about 195 mg, about 55 mg to about 190 mg, about 55 mg to about 185 mg, about 55 mg to about 180 mg, about 55 mg to about 175 mg, about 55 mg to about 170 mg, about 55 mg to about 165 mg, about 55 mg to about 160 mg, about 55 mg to about 155 mg, about 55 mg to about 150 mg, about 55 mg to about 145 mg, about 55 mg to about 140 mg, about 55 mg to about 135 mg, about 55 mg to about 130 mg, about 55 mg to about 125 mg, about 55 mg to about 120 mg, about 55 mg to about 115 mg, about 55 mg to about 110 mg, about 55 mg to about 105 mg, about 55 mg to about 100 mg, about 55 mg to about 95 mg, about 55 mg to about 90 mg, about 55 mg to about 85 mg, about 55 mg to about 80 mg, about 55 mg to about 75 mg, about 55 mg to about 70 mg, about 55 mg to about 65 mg, about 55 mg to about 60 mg, about 60 mg to about 200 mg, about 60 mg to about 195 mg, about 60 mg to about 190 mg, about 60 mg to about 185 mg, about 60 mg to about 180 mg, about 60 mg to about 175 mg, about 60 mg to about 170 mg, about 60 mg to about 165 mg, about 60 mg to about 160 mg, about 60 mg to about 155 mg, about 60 mg to about 150 mg, about 60 mg to about 145 mg, about 60 mg to about 140 mg, about 60 mg to about 135 mg, about 60 mg to about 130 mg, about 60 mg to about 125 mg, about 60 mg to about 120 mg, about 60 mg to about 115 mg, about 60 mg to about 110 mg, about 60 mg to about 105 mg, about 60 mg to about 100 mg, about 60 mg to about 95 mg, about 60 mg to about 90 mg, about 60 mg to about 85 mg, about 60 mg to about 80 mg, about 60 mg to about 75 mg, about 60 mg to about 70 mg, about 60 mg to about 65 mg, about 65 mg to about 200 mg, about 65 mg to about 195 mg, about 65 mg to about 190 mg, about 65 mg to about 185 mg, about 65 mg to about 180 mg, about 65 mg to about 175 mg, about 65 mg to about 170 mg, about 65 mg to about 165 mg, about 65 mg to about 160 mg, about 65 mg to about 155 mg, about 65 mg to about 150 mg, about 65 mg to about 145 mg, about 65 mg to about 140 mg, about 65 mg to about 135 mg, about 65 mg to about 130 mg, about 65 mg to about 125 mg, about 65 mg to about 120 mg, about 65 mg to about 115 mg, about 65 mg to about 110 mg, about 65 mg to about 105 mg, about 65 mg to about 100 mg, about 65 mg to about 95 mg, about 65 mg to about 90 mg, about 65 mg to about 85 mg, about 65 mg to about 80 mg, about 65 mg to about 75 mg, about 65 mg to about 70 mg, about 70 mg to about 200 mg, about 70 mg to about 195 mg, about 70 mg to about 190 mg, about 70 mg to about 185 mg, about 70 mg to about 180 mg, about 70 mg to about 175 mg, about 70 mg to about 170 mg, about 70 mg to about 165 mg, about 70 mg to about 160 mg, about 70 mg to about 155 mg, about 70 mg to about 150 mg, about 70 mg to about 145 mg, about 70 mg to about 140 mg, about 70 mg to about 135 mg, about 70 mg to about 130 mg, about 70 mg to about 125 mg, about 70 mg to about 120 mg, about 70 mg to about 115 mg, about 70 mg to about 110 mg, about 70 mg to about 105 mg, about 70 mg to about 100 mg, about 70 mg to about 95 mg, about 70 mg to about 90 mg, about 70 mg to about 85 mg, about 70 mg to about 80 mg, about 70 mg to about 75 mg, about 75 mg to about 200 mg, about 75 mg to about 195 mg, about 75 mg to about 190 mg, about 75 mg to about 185 mg, about 75 mg to about 180 mg, about 75 mg to about 175 mg, about 75 mg to about 170 mg, about 75 mg to about 165 mg, about 75 mg to about 160 mg, about 75 mg to about 155 mg, about 75 mg to about 150 mg, about 75 mg to about 145 mg, about 75 mg to about 140 mg, about 75 mg to about 135 mg, about 75 mg to about 130 mg, about 75 mg to about 125 mg, about 75 mg to about 120 mg, about 75 mg to about 115 mg, about 75 mg to about 110 mg, about 75 mg to about 105 mg, about 75 mg to about 100 mg, about 75 mg to about 95 mg, about 75 mg to about 90 mg, about 75 mg to about 85 mg, about 75 mg to about 80 mg, about 80 mg to about 200 mg, about 80 mg to about 195 mg, about 80 mg to about 190 mg, about 80 mg to about 185 mg, about 80 mg to about 180 mg, about 80 mg to about 175 mg, about 80 mg to about 170 mg, about 80 mg to about 165 mg, about 80 mg to about 160 mg, about 80 mg to about 155 mg, about 80 mg to about 150 mg, about 80 mg to about 145 mg, about 80 mg to about 140 mg, about 80 mg to about 135 mg, about 80 mg to about 130 mg, about 80 mg to about 125 mg, about 80 mg to about 120 mg, about 80 mg to about 115 mg, about 80 mg to about 110 mg, about 80 mg to about 105 mg, about 80 mg to about 100 mg, about 80 mg to about 95 mg, about 80 mg to about 90 mg, about 80 mg to about 85 mg, about 85 mg to about 200 mg, about 85 mg to about 195 mg, about 85 mg to about 190 mg, about 85 mg to about 185 mg, about 85 mg to about 180 mg, about 85 mg to about 175 mg, about 85 mg to about 170 mg, about 85 mg to about 165 mg, about 85 mg to about 160 mg, about 85 mg to about 155 mg, about 85 mg to about 150 mg, about 85 mg to about 145 mg, about 85 mg to about 140 mg, about 85 mg to about 135 mg, about 85 mg to about 130 mg, about 85 mg to about 125 mg, about 85 mg to about 120 mg, about 85 mg to about 115 mg, about 85 mg to about 110 mg, about 85 mg to about 105 mg, about 85 mg to about 100 mg, about 85 mg to about 95 mg, about 85 mg to about 90 mg, about 90 mg to about 200 mg, about 90 mg to about 195 mg, about 90 mg to about 190 mg, about 90 mg to about 185 mg, about 90 mg to about 180 mg, about 90 mg to about 175 mg, about 90 mg to about 170 mg, about 90 mg to about 165 mg, about 90 mg to about 160 mg, about 90 mg to about 155 mg, about 90 mg to about 150 mg, about 90 mg to about 145 mg, about 90 mg to about 140 mg, about 90 mg to about 135 mg, about 90 mg to about 130 mg, about 90 mg to about 125 mg, about 90 mg to about 120 mg, about 90 mg to about 115 mg, about 90 mg to about 110 mg, about 90 mg to about 105 mg, about 90 mg to about 100 mg, about 90 mg to about 95 mg, about 95 mg to about 200 mg, about 95 mg to about 195 mg, about 95 mg to about 190 mg, about 95 mg to about 185 mg, about 95 mg to about 180 mg, about 95 mg to about 175 mg, about 95 mg to about 170 mg, about 95 mg to about 165 mg, about 95 mg to about 160 mg, about 95 mg to about 155 mg, about 95 mg to about 150 mg, about 95 mg to about 145 mg, about 95 mg to about 140 mg, about 95 mg to about 135 mg, about 95 mg to about 130 mg, about 95 mg to about 125 mg, about 95 mg to about 120 mg, about 95 mg to about 115 mg, about 95 mg to about 110 mg, about 95 mg to about 105 mg, about 95 mg to about 100 mg, about 100 mg to about 200 mg, about 100 mg to about 195 mg, about 100 mg to about 190 mg, about 100 mg to about 185 mg, about 100 mg to about 180 mg, about 100 mg to about 175 mg, about 100 mg to about 170 mg, about 100 mg to about 165 mg, about 100 mg to about 160 mg, about 100 mg to about 155 mg, about 100 mg to about 150 mg, about 100 mg to about 145 mg, about 100 mg to about 140 mg, about 100 mg to about 135 mg, about 100 mg to about 130 mg, about 100 mg to about 125 mg, about 100 mg to about 120 mg, about 100 mg to about 115 mg, about 100 mg to about 110 mg, about 100 mg to about 105 mg, about 105 mg to about 200 mg, about 105 mg to about 195 mg, about 105 mg to about 190 mg, about 105 mg to about 185 mg, about 105 mg to about 180 mg, about 105 mg to about 175 mg, about 105 mg to about 170 mg, about 105 mg to about 165 mg, about 105 mg to about 160 mg, about 105 mg to about 155 mg, about 105 mg to about 150 mg, about 105 mg to about 145 mg, about 105 mg to about 140 mg, about 105 mg to about 135 mg, about 105 mg to about 130 mg, about 105 mg to about 125 mg, about 105 mg to about 120 mg, about 105 mg to about 115 mg, about 105 mg to about 110 mg, about 110 mg to about 200 mg, about 110 mg to about 195 mg, about 110 mg to about 190 mg, about 110 mg to about 185 mg, about 110 mg to about 180 mg, about 110 mg to about 175 mg, about 110 mg to about 170 mg, about 110 mg to about 165 mg, about 110 mg to about 160 mg, about 110 mg to about 155 mg, about 110 mg to about 150 mg, about 110 mg to about 145 mg, about 110 mg to about 140 mg, about 110 mg to about 135 mg, about 110 mg to about 130 mg, about 110 mg to about 125 mg, about 110 mg to about 120 mg, about 110 mg to about 115 mg, about 115 mg to about 200 mg, about 115 mg to about 195 mg, about 115 mg to about 190 mg, about 115 mg to about 185 mg, about 115 mg to about 180 mg, about 115 mg to about 175 mg, about 115 mg to about 170 mg, about 115 mg to about 165 mg, about 115 mg to about 160 mg, about 115 mg to about 155 mg, about 115 mg to about 150 mg, about 115 mg to about 145 mg, about 115 mg to about 140 mg, about 115 mg to about 135 mg, about 115 mg to about 130 mg, about 115 mg to about 125 mg, about 115 mg to about 120 mg, about 120 mg to about 200 mg, about 120 mg to about 195 mg, about 120 mg to about 190 mg, about 120 mg to about 185 mg, about 120 mg to about 180 mg, about 120 mg to about 175 mg, about 120 mg to about 170 mg, about 120 mg to about 165 mg, about 120 mg to about 160 mg, about 120 mg to about 155 mg, about 120 mg to about 150 mg, about 120 mg to about 145 mg, about 120 mg to about 140 mg, about 120 mg to about 135 mg, about 120 mg to about 130 mg, about 120 mg to about 125 mg, about 125 mg to about 200 mg, about 125 mg to about 195 mg, about 125 mg to about 190 mg, about 125 mg to about 185 mg, about 125 mg to about 180 mg, about 125 mg to about 175 mg, about 125 mg to about 170 mg, about 125 mg to about 165 mg, about 125 mg to about 160 mg, about 125 mg to about 155 mg, about 125 mg to about 150 mg, about 125 mg to about 145 mg, about 125 mg to about 140 mg, about 125 mg to about 135 mg, about 125 mg to about 130 mg, about 130 mg to about 200 mg, about 130 mg to about 195 mg, about 130 mg to about 190 mg, about 130 mg to about 185 mg, about 130 mg to about 180 mg, about 130 mg to about 175 mg, about 130 mg to about 170 mg, about 130 mg to about 165 mg, about 130 mg to about 160 mg, about 130 mg to about 155 mg, about 130 mg to about 150 mg, about 130 mg to about 145 mg, about 130 mg to about 140 mg, about 130 mg to about 135 mg, about 135 mg to about 200 mg, about 135 mg to about 195 mg, about 135 mg to about 190 mg, about 135 mg to about 185 mg, about 135 mg to about 180 mg, about 135 mg to about 175 mg, about 135 mg to about 170 mg, about 135 mg to about 165 mg, about 135 mg to about 160 mg, about 135 mg to about 155 mg, about 135 mg to about 150 mg, about 135 mg to about 145 mg, about 135 mg to about 140 mg, about 140 mg to about 200 mg, about 140 mg to about 195 mg, about 140 mg to about 190 mg, about 140 mg to about 185 mg, about 140 mg to about 180 mg, about 140 mg to about 175 mg, about 140 mg to about 170 mg, about 140 mg to about 165 mg, about 140 mg to about 160 mg, about 140 mg to about 155 mg, about 140 mg to about 150 mg, about 140 mg to about 145 mg, about 145 mg to about 200 mg, about 145 mg to about 195 mg, about 145 mg to about 190 mg, about 145 mg to about 185 mg, about 145 mg to about 180 mg, about 145 mg to about 175 mg, about 145 mg to about 170 mg, about 145 mg to about 165 mg, about 145 mg to about 160 mg, about 145 mg to about 155 mg, about 145 mg to about 150 mg, about 150 mg to about 200 mg, about 150 mg to about 195 mg, about 150 mg to about 190 mg, about 150 mg to about 185 mg, about 150 mg to about 180 mg, about 150 mg to about 175 mg, about 150 mg to about 170 mg, about 150 mg to about 165 mg, about 150 mg to about 160 mg, about 150 mg to about 155 mg, about 155 mg to about 200 mg, about 155 mg to about 195 mg, about 155 mg to about 190 mg, about 155 mg to about 185 mg, about 155 mg to about 180 mg, about 155 mg to about 175 mg, about 155 mg to about 170 mg, about 155 mg to about 165 mg, about 155 mg to about 160 mg, about 160 mg to about 200 mg, about 160 mg to about 195 mg, about 160 mg to about 190 mg, about 160 mg to about 185 mg, about 160 mg to about 180 mg, about 160 mg to about 175 mg, about 160 mg to about 170 mg, about 160 mg to about 165 mg, about 165 mg to about 200 mg, about 165 mg to about 195 mg, about 165 mg to about 190 mg, about 165 mg to about 185 mg, about 165 mg to about 180 mg, about 165 mg to about 175 mg, about 165 mg to about 170 mg, about 170 mg to about 200 mg, about 170 mg to about 195 mg, about 170 mg to about 190 mg, about 170 mg to about 185 mg, about 170 mg to about 180 mg, about 170 mg to about 175 mg, about 175 mg to about 200 mg, about 175 mg to about 195 mg, about 175 mg to about 190 mg, about 175 mg to about 185 mg, about 175 mg to about 180 mg, about 180 mg to about 200 mg, about 180 mg to about 195 mg, about 180 mg to about 190 mg, about 180 mg to about 185 mg, about 185 mg to about 200 mg, about 185 mg to about 195 mg, about 185 mg to about 190 mg, about 190 mg to about 200 mg, about 190 mg to about 195 mg, or about 195 mg to about 200 mg.

[1649] In some embodiments, the amount of the immune modulator that is administered is less than an amount that is effective when the immune modulator is delivered systemically.

[1650] In some embodiments, the amount of the immune modulator that is administered is an induction dose. In some embodiments, such induction dose is effective to induce remission of the TNF and cytokine storm and healing of acute inflammation and lesions. In some embodiments, the induction dose is administered once a day. In some embodiments of any of the methods described herein, the induction dose is administered once every two days. In some embodiments, the induction dose is administered once every three days. In some embodiments, the induction dose is administered once a week. In some embodiments, the induction dose is administered once a day, once every three days, or once a week, over a period of about 6-8 weeks.

[1651] In some embodiments, the method comprises administering (i) an amount of the immune modulator that is an induction dose, and (ii) an amount of the immune modulator that is a maintenance dose, in this order. In some embodiments, step (ii) is repeated one or more times. In some embodiments, the induction dose is equal to the maintenance dose. In some embodiments, the induction dose is greater than the maintenance dose. In some embodiments, the induction dose is five times greater than the maintenance dose. In some embodiments, the induction dose is two times greater than the maintenance dose.

[1652] In some embodiments, the induction dose is the same as or higher than an induction dose administered systemically for treatment of the same disorder to a subject. In more particular embodiments, the induction dose is the same as or higher than an induction dose administered systemically for treatment of the same disorder to a subject, and the maintenance dose is lower than the maintenance dose administered systemically for treatment of the same disorder to a subject. In some embodiments, the induction dose is the same as or higher than an induction dose administered systemically for treatment of the same disorder to a subject, and the maintenance dose is higher than the maintenance dose administered systemically for treatment of the same disorder to a subject.

[1653] In some embodiments an induction dose of the immune modulator and a maintenance dose of immune modulator are each administered to the subject by administering a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator, wherein the pharmaceutical composition is a device. In some embodiments an induction dose of immune modulator is administered to the subject in a different manner from the maintenance dose. As an example, the induction dose may be administered systemically. In some embodiments, the induction dose may be administered other than orally. As an example, the induction dose may be administered rectally. As an example, the induction dose may be administered intravenously. As an example, the induction dose may be administered subcutaneously. In some embodiments, the induction dose may be administered by spray catheter.

[1654] In some embodiments, the concentration of the immune modulator delivered at the location in the gastrointestinal tract is 10%, 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 1000%, 2000% greater than the concentration of the immune modulator in plasma.

[1655] In some embodiments, the method provides a concentration of the immune modulator at a location that is the intended site of release is 2-100 times greater than the concentration at a location that is not the intended site of release in the GI tract.

[1656] In some embodiments, the method comprises delivering the immune modulator at the location in the gastrointestinal tract as a single bolus.

[1657] In some embodiments, the method comprises delivering the immune modulator at the location in the gastrointestinal tract as more than one bolus.

[1658] In some embodiments, the method comprises delivering the immune modulator at the location in the gastrointestinal tract in a continuous manner.

[1659] In some embodiments, the method comprises delivering the immune modulator at the location in the gastrointestinal tract over a time period of 20 or more minutes.

[1660] In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 10 pg/ml. In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 3 pg/ml. In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 1 pg/ml. In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.3 pg/ml. In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.1 pg/ml. In some embodiments, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.01 pg/ml. In some embodiments, the values of the concentration of the immune modulator in the plasma of the subject provided herein refer to C.sub.trough, that is, the lowest value of the concentration prior to administration of the next dose.

[1661] In some embodiments, the method provides a concentration of the immune modulator inhibitor in the plasma of the subject that is, e.g., about 1 ng/L to about 100 ng/mL, about 1 ng/mL to about 95 ng/mL, about 1 ng/mL to about 90 ng/mL, about 1 ng/mL to about 85 ng/mL, about 1 ng/mL to about 80 ng/mL, about 1 ng/mL to about 75 ng/mL, about 1 ng/mL to about 70 ng/mL, about 1 ng/mL to about 65 ng/mL, about 1 ng/mL to about 60 ng/mL, about 1 ng/mL to about 55 ng/mL, about 1 ng/mL to about 50 ng/mL, about 1 ng/mL to about 45 ng/mL, about 1 ng/mL to about 40 ng/mL, about 1 ng/mL to about 35 ng/mL, about 1 ng/mL to about 30 ng/mL, about 1 ng/mL to about 25 ng/mL, about 1 ng/mL to about 20 ng/mL, about 1 ng/mL to about 15 ng/mL, about 1 ng/mL to about 10 ng/mL, about 1 ng/mL to about 5 ng/mL, about 2 ng/L to about 100 ng/mL, about 2 ng/mL to about 95 ng/mL, about 2 ng/mL to about 90 ng/mL, about 2 ng/mL to about 85 ng/mL, about 2 ng/mL to about 80 ng/mL, about 2 ng/mL to about 75 ng/mL, about 2 ng/mL to about 70 ng/mL, about 2 ng/mL to about 65 ng/mL, about 2 ng/mL to about 60 ng/mL, about 2 ng/mL to about 55 ng/mL, about 2 ng/mL to about 50 ng/mL, about 2 ng/mL to about 45 ng/mL, about 2 ng/mL to about 40 ng/mL, about 2 ng/mL to about 35 ng/mL, about 2 ng/mL to about 30 ng/mL, about 2 ng/mL to about 25 ng/mL, about 2 ng/mL to about 20 ng/mL, about 2 ng/mL to about 15 ng/mL, about 2 ng/mL to about 10 ng/mL, about 2 ng/mL to about 5 ng/mL, about 5 ng/L to about 100 ng/mL, about 5 ng/mL to about 95 ng/mL, about 5 ng/mL to about 90 ng/mL, about 5 ng/mL to about 85 ng/mL, about 5 ng/mL to about 80 ng/mL, about 5 ng/mL to about 75 ng/mL, about 5 ng/mL to about 70 ng/mL, about 5 ng/mL to about 65 ng/mL, about 5 ng/mL to about 60 ng/mL, about 5 ng/mL to about 55 ng/mL, about 5 ng/mL to about 50 ng/mL, about 5 ng/mL to about 45 ng/mL, about 5 ng/mL to about 40 ng/mL, about 5 ng/mL to about 35 ng/mL, about 5 ng/mL to about 30 ng/mL, about 5 ng/mL to about 25 ng/mL, about 5 ng/mL to about 20 ng/mL, about 5 ng/mL to about 15 ng/mL, about 5 ng/mL to about 10 ng/mL, about 10 ng/L to about 100 ng/mL, about 10 ng/mL to about 95 ng/mL, about 10 ng/mL to about 90 ng/mL, about 10 ng/mL to about 85 ng/mL, about 10 ng/mL to about 80 ng/mL, about 10 ng/mL to about 75 ng/mL, about 10 ng/mL to about 70 ng/mL, about 10 ng/mL to about 65 ng/mL, about 10 ng/mL to about 60 ng/mL, about 10 ng/mL to about 55 ng/mL, about 10 ng/mL to about 50 ng/mL, about 10 ng/mL to about 45 ng/mL, about 10 ng/mL to about 40 ng/mL, about 10 ng/mL to about 35 ng/mL, about 10 ng/mL to about 30 ng/mL, about 10 ng/mL to about 25 ng/mL, about 10 ng/mL to about 20 ng/mL, about 10 ng/mL to about 15 ng/mL, about 15 ng/L to about 100 ng/mL, about 15 ng/mL to about 95 ng/mL, about 15 ng/mL to about 90 ng/mL, about 15 ng/mL to about 85 ng/mL, about 15 ng/mL to about 80 ng/mL, about 15 ng/mL to about 75 ng/mL, about 15 ng/mL to about 70 ng/mL, about 15 ng/mL to about 65 ng/mL, about 15 ng/mL to about 60 ng/mL, about 15 ng/mL to about 55 ng/mL, about 15 ng/mL to about 50 ng/mL, about 15 ng/mL to about 45 ng/mL, about 15 ng/mL to about 40 ng/mL, about 15 ng/mL to about 35 ng/mL, about 15 ng/mL to about 30 ng/mL, about 15 ng/mL to about 25 ng/mL, about 15 ng/mL to about 20 ng/mL, about 20 ng/L to about 100 ng/mL, about 20 ng/mL to about 95 ng/mL, about 20 ng/mL to about 90 ng/mL, about 20 ng/mL to about 85 ng/mL, about 20 ng/mL to about 80 ng/mL, about 20 ng/mL to about 75 ng/mL, about 20 ng/mL to about 70 ng/mL, about 20 ng/mL to about 65 ng/mL, about 20 ng/mL to about 60 ng/mL, about 20 ng/mL to about 55 ng/mL, about 20 ng/mL to about 50 ng/mL, about 20 ng/mL to about 45 ng/mL, about 20 ng/mL to about 40 ng/mL, about 20 ng/mL to about 35 ng/mL, about 20 ng/mL to about 30 ng/mL, about 20 ng/mL to about 25 ng/mL, about 25 ng/L to about 100 ng/mL, about 25 ng/mL to about 95 ng/mL, about 25 ng/mL to about 90 ng/mL, about 25 ng/mL to about 85 ng/mL, about 25 ng/mL to about 80 ng/mL, about 25 ng/mL to about 75 ng/mL, about 25 ng/mL to about 70 ng/mL, about 25 ng/mL to about 65 ng/mL, about 25 ng/mL to about 60 ng/mL, about 25 ng/mL to about 55 ng/mL, about 25 ng/mL to about 50 ng/mL, about 25 ng/mL to about 45 ng/mL, about 25 ng/mL to about 40 ng/mL, about 25 ng/mL to about 35 ng/mL, about 25 ng/mL to about 30 ng/mL, about 30 ng/L to about 100 ng/mL, about 30 ng/mL to about 95 ng/mL, about 30 ng/mL to about 90 ng/mL, about 30 ng/mL to about 85 ng/mL, about 30 ng/mL to about 80 ng/mL, about 30 ng/mL to about 75 ng/mL, about 30 ng/mL to about 70 ng/mL, about 30 ng/mL to about 65 ng/mL, about 30 ng/mL to about 60 ng/mL, about 30 ng/mL to about 55 ng/mL, about 30 ng/mL to about 50 ng/mL, about 30 ng/mL to about 45 ng/mL, about 30 ng/mL to about 40 ng/mL, about 30 ng/mL to about 35 ng/mL, about 35 ng/L to about 100 ng/mL, about 35 ng/mL to about 95 ng/mL, about 35 ng/mL to about 90 ng/mL, about 35 ng/mL to about 85 ng/mL, about 35 ng/mL to about 80 ng/mL, about 35 ng/mL to about 75 ng/mL, about 35 ng/mL to about 70 ng/mL, about 35 ng/mL to about 65 ng/mL, about 35 ng/mL to about 60 ng/mL, about 35 ng/mL to about 55 ng/mL, about 35 ng/mL to about 50 ng/mL, about 35 ng/mL to about 45 ng/mL, about 35 ng/mL to about 40 ng/mL, about 40 ng/L to about 100 ng/mL, about 40 ng/mL to about 95 ng/mL, about 40 ng/mL to about 90 ng/mL, about 40 ng/mL to about 85 ng/mL, about 40 ng/mL to about 80 ng/mL, about 40 ng/mL to about 75 ng/mL, about 40 ng/mL to about 70 ng/mL, about 40 ng/mL to about 65 ng/mL, about 40 ng/mL to about 60 ng/mL, about 40 ng/mL to about 55 ng/mL, about 40 ng/mL to about 50 ng/mL, about 40 ng/mL to about 45 ng/mL, about 45 ng/L to about 100 ng/mL, about 45 ng/mL to about 95 ng/mL, about 45 ng/mL to about 90 ng/mL, about 45 ng/mL to about 85 ng/mL, about 45 ng/mL to about 80 ng/mL, about 45 ng/mL to about 75 ng/mL, about 45 ng/mL to about 70 ng/mL, about 45 ng/mL to about 65 ng/mL, about 45 ng/mL to about 60 ng/mL, about 45 ng/mL to about 55 ng/mL, about 45 ng/mL to about 50 ng/mL, about 50 ng/L to about 100 ng/mL, about 50 ng/mL to about 95 ng/mL, about 50 ng/mL to about 90 ng/mL, about 50 ng/mL to about 85 ng/mL, about 50 ng/mL to about 80 ng/mL, about 50 ng/mL to about 75 ng/mL, about 50 ng/mL to about 70 ng/mL, about 50 ng/mL to about 65 ng/mL, about 50 ng/mL to about 60 ng/mL, about 50 ng/mL to about 55 ng/mL, about 55 ng/L to about 100 ng/mL, about 55 ng/mL to about 95 ng/mL, about 55 ng/mL to about 90 ng/mL, about 55 ng/mL to about 85 ng/mL, about 55 ng/mL to about 80 ng/mL, about 55 ng/mL to about 75 ng/mL, about 55 ng/mL to about 70 ng/mL, about 55 ng/mL to about 65 ng/mL, about 55 ng/mL to about 60 ng/mL, about 60 ng/L to about 100 ng/mL, about 60 ng/mL to about 95 ng/mL, about 60 ng/mL to about 90 ng/mL, about 60 ng/mL to about 85 ng/mL, about 60 ng/mL to about 80 ng/mL, about 60 ng/mL to about 75 ng/mL, about 60 ng/mL to about 70 ng/mL, about 60 ng/mL to about 65 ng/mL, about 65 ng/L to about 100 ng/mL, about 65 ng/mL to about 95 ng/mL, about 65 ng/mL to about 90 ng/mL, about 65 ng/mL to about 85 ng/mL, about 65 ng/mL to about 80 ng/mL, about 65 ng/mL to about 75 ng/mL, about 65 ng/mL to about 70 ng/mL, about 70 ng/L to about 100 ng/mL, about 70 ng/mL to about 95 ng/mL, about 70 ng/mL to about 90 ng/mL, about 70 ng/mL to about 85 ng/mL, about 70 ng/mL to about 80 ng/mL, about 70 ng/mL to about 75 ng/mL, about 75 ng/L to about 100 ng/mL, about 75 ng/mL to about 95 ng/mL, about 75 ng/mL to about 90 ng/mL, about 75 ng/mL to about 85 ng/mL, about 75 ng/mL to about 80 ng/mL, about 80 ng/L to about 100 ng/mL, about 80 ng/mL to about 95 ng/mL, about 80 ng/mL to about 90 ng/mL, about 80 ng/mL to about 85 ng/mL, about 85 ng/L to about 100 ng/mL, about 85 ng/mL to about 95 ng/mL, about 85 ng/mL to about 90 ng/mL, about 90 ng/L to about 100 ng/mL, about 90 ng/mL to about 95 ng/mL, or about 95 ng/mL to about 100 ng/mL.

[1662] In some embodiments, the method provides a concentration C.sub.max of the immune modulator in the plasma of the subject that is less than 10 pg/ml. In some embodiments, the method provides a concentration C.sub.max of the immune modulator in the plasma of the subject that is less than 3 g/ml. In some embodiments, the method provides a concentration Cm of the immune modulator in the plasma of the subject that is less than 1 pg/ml. In some embodiments, the method provides a concentration C.sub.max of the immune modulator in the plasma of the subject that is less than 0.3 pg/ml. In some embodiments, the method provides a concentration C.sub.max of the immune modulator in the plasma of the subject that is less than 0.1 pg/ml. In some embodiments, the method provides a concentration C.sub.max of the immune modulator in the plasma of the subject that is less than 0.01 pg/ml.

[1663] In some more particular embodiments, a method of treating a disease or condition of the gastrointestinal tract of a subject comprises administering an induction dose and subsequently a maintenance dose of the immune modulator. In some more particular embodiments, the total induction dose for a given period of time is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times or at least 10 times greater than a systemic induction dose for the same period of time. In some more particular embodiments, the total induction dose for a 2 week period is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times or at least 10 times greater than a systemic induction dose for the same period of time. In some more particular embodiments, the total induction dose for a 4 week period is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times or at least 10 times greater than a systemic induction dose for the same period of time. In some more particular embodiments, the total induction dose for a 6 week period is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times or at least 10 times greater than a systemic induction dose for the same period of time. In some more particular embodiments, the total induction dose for a 8 week period is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 8 times or at least 10 times greater than a systemic induction dose for the same period of time.

[1664] In some more particular embodiments, an ingestible device comprising an immune modulatory agent may be administered once per day or more than once per day, for example, 1, 2, 3, 4 or more times per day. In some more particular embodiments, two or more ingestible devices may be administered at the same time. In some more particular embodiments, two or more ingestible devices may be administered 1 minute apart, 2 minutes apart, 3 minutes apart, 4 minutes apart, 5 minutes apart, 10 minutes apart, 15 minutes apart, 30 minutes apart, or 60 minutes apart. In some more particular embodiments, two or more ingestible devices may be administered 1 hour apart, 2 hours apart, 3 hours apart, 4 hours apart, 5 hours apart, 6 hours apart, 7 hours apart, 8 hours apart, 9 hours apart, 10 hours apart, 11 hours apart, or 12 hours apart.

[1665] In some more particular embodiments, administration of an immune modulator using any of the devices or compositions described herein can provide a reduction in TH memory cell count in the mesenteric lymph nodes of the subject relative to systemic administration of the same amount of the immune modulator that is at least a 10% reduction, at least a 20% reduction, at least a 30% reduction, at least a 40% reduction or at least a 50% reduction.

[1666] In some more particular embodiments, administration of an immune modulator using any of the devices or compositions described herein can provide a reduction in TH memory cell count in the Peyer's Patches of the subject relative to systemic administration of the same amount of the immune modulator that is at least a 10% reduction.

[1667] In some more particular embodiments, administration of an immune modulator using any of the devices or compositions described herein can provide an increase in TH memory cell count in the blood of the subject relative to systemic administration of the same amount of the immune modulator that is at least a 1% increase, at least a 5% increase, at least at 10% increase or at least a 15% increase.

[1668] In some embodiments, the method does not comprise delivering an immune modulator rectally to the subject.

[1669] In some embodiments, the method does not comprise delivering an immune modulator via an enema to the subject.

[1670] In some embodiments, the method does not comprise delivering an immune modulator via suppository to the subject.

[1671] In some embodiments, the method does not comprise delivering an immune modulator via instillation to the rectum of a subject.

[1672] In some embodiments, the methods disclosed herein comprise producing a therapeutically effective degradation product of the immune modulator in the gastrointestinal tract. In some embodiments, a therapeutically effective amount of the degradation product is produced.

[1673] In some embodiments, the methods comprising administering the immune modulator in the manner disclosed herein disclosed herein result in a reduced immunosuppressive properties relative to methods of administration of the immune modulator systemically.

[1674] In some embodiments, the methods comprising administering the immune modulator in the manner disclosed herein disclosed herein result in reduced immunogenicity relative to methods of administration of the immune modulator systemically.

Patients Condition, Diagnosis and Treatment

[1675] In some embodiments herein, the method of treating an inflammatory disease or disorder that arises in a tissue that originates from the endoderm that comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to an intended site of release comprises one or more of the following: [1676] a) identifying a subject having an inflammatory disease or condition that arises in tissue originating from the endoderm; [1677] b) determination of the severity of the disease; and [1678] c) evaluating the subject for suitability to treatment, for example by determining the patency of the subject's GI tract, or if the patients has strictures or fistulae; [1679] d) administration of an induction dose or of a maintenance dose of an immune modulator; [1680] e) monitoring the progress of the disease one or more times, for example over a period of about 1-14 weeks, such as about 6-8 weeks following administration of the immune modulator, including at the 6-8 week time point, or over a period of about 52 weeks following administration of the immune modulator, including at the 52 week time point.

[1681] As used herein, an induction dose is a dose of drug that may be administered, for example, at the beginning of a course of treatment, and that is higher than the maintenance dose administered during treatment. An induction dose may also be administered during treatment, for example if the condition of the patients becomes worse.

[1682] As used herein, a maintenance dose is a dose of drug that is provided on a repetitive basis, for example at regular dosing intervals.

[1683] In some embodiments the immune modulator is released from an ingestible device.

[1684] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises a) hereinabove.

[1685] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises b) hereinabove.

[1686] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises c) hereinabove.

[1687] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises d) hereinabove.

[1688] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises e) hereinabove.

[1689] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises a) and b) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises a) and c) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises a) and d) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises a) and e) hereinabove.

[1690] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises b) and c) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises b) and d) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises b) and e) hereinabove.

[1691] In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises c) and d) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises c) and e) hereinabove. In some embodiments herein, the method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm comprises releasing an immune modulator at a location in the gastrointestinal tract that is proximate to the intended site of release comprises d) and e) hereinabove.

[1692] In some embodiments, one or more steps a) to e) herein comprise endoscopy of the gastrointestinal tract. In some embodiments, one or more steps a) to e) herein comprise colonoscopy of the gastrointestinal tract. In some embodiments, one or more steps a) to e) herein is performed one or more times. In some embodiments, such one or more of such one or more steps a) to e) is performed after releasing the immune modulator at the location in the gastrointestinal tract that is proximate to the intended site of release.

[1693] In some embodiments, the method comprises administering one or more maintenance doses following administration of the induction dose. In some embodiments an induction dose of an immune modulator and a maintenance dose of an immune modulator are each administered to the subject by administering a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator. In some embodiments an induction dose of an immune modulator is administered to the subject in a different manner from the maintenance dose. As an example, the maintenance dose may be administered systemically, while the maintenance dose is administered locally using a device. In one embodiment, a maintenance dose is administered systemically, and an induction dose is administered using a device every 1, 2, 3, 4, 5, 6, 7, 10, 15, 20, 25, 30, 35, 40, or 45 days. In another embodiment, a maintenance dose is administered systemically, and an induction dose is administered when a disease flare up is detected or suspected.

[1694] In some embodiments, the induction dose is a dose of the immune modulator is administered in an ingestible device as disclosed herein. In some embodiments, the maintenance dose is a dose of the immune modulator administered in an ingestible device as disclosed herein.

[1695] In some embodiments, the induction dose is a dose of the immune modulator administered in an ingestible device as disclosed herein. In some embodiments, the maintenance dose is a dose of the immune modulator delivered systemically, such as orally with a tablet or capsule, or subcutaneously, or intravenously.

[1696] In some embodiments, the induction dose is a dose of the immune modulator delivered systemically, such as orally with a tablet or capsule, or subcutaneously, or intravenously. In some embodiments, the maintenance dose is a dose of the immune modulator administered in an ingestible device as disclosed herein.

[1697] In some embodiments, the induction dose is a dose of the immune modulator administered in an ingestible device as disclosed herein. In some embodiments, the maintenance dose is a dose of a second agent as disclosed herein delivered systemically, such as orally with a tablet or capsule, or subcutaneously, or intravenously.

[1698] In some embodiments, the induction dose is a dose of a second agent as disclosed herein delivered systemically, such as orally with a tablet or capsule, or subcutaneously, or intravenously. In some embodiments, the maintenance dose is a dose of the immune modulator administered in an ingestible device as disclosed herein.

[1699] In one embodiment of the methods provided herein, the patient is not previously treated with an immune modulator.

[1700] In some embodiments, the method comprises identifying the intended site of release substantially at the same time as releasing the immune modulator.

[1701] In some embodiments, the method comprises monitoring the progress of the disease. In some embodiments, the method comprises administering an immune modulator with a spray catheter. For example, administering an immune modulator with a spray catheter may be performed in step (e) hereinabove.

[1702] In some embodiments, the method does not comprise administering an immune modulator with a spray catheter.

[1703] In some embodiments, data obtained from cell culture assays and animal studies can be used in formulating an appropriate dosage of any given immune modulator. The effectiveness and dosing of any immune modulator can be determined by a health care professional or veterinary professional using methods known in the art, as well as by the observation of one or more disease symptoms in a subject (e.g., a human). Certain factors may influence the dosage and timing required to effectively treat a subject (e.g., the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and the presence of other diseases).

[1704] In some embodiments, the subject is further administered an additional therapeutic agent (e.g., any of the additional therapeutic agents described herein). The additional therapeutic agent can be administered to the subject at substantially the same time as the immune modulator or pharmaceutical composition comprising it is administered and/or at one or more other time points. In some embodiments, the additional therapeutic agent is formulated together with the immune modulator (e.g., using any of the examples of formulations described herein).

[1705] In some embodiments, the subject is administered a dose of the immune modulator at least once a month (e.g., at least twice a month, at least three times a month, at least four times a month, at least once a week, at least twice a week, three times a week, once a day, or twice a day). The immune modulator may be administered to a subject chronically. Chronic treatments include any form of repeated administration for an extended period of time, such as repeated administrations for one or more months, between a month and a year, one or more years, more than five years, more than 10 years, more than 15 years, more than 20 years, more than 25 years, more than 30 years, more than 35 years, more than 40 years, more than 45 years, or longer. Alternatively or in addition, chronic treatments may be administered. Chronic treatments can involve regular administrations, for example one or more times a day, one or more times a week, or one or more times a month. For example, chronic treatment can include administration (e.g., intravenous administration) about every two weeks (e.g., between about every 10 to 18 days).

[1706] A suitable dose may be the amount that is the lowest dose effective to produce a desired therapeutic effect. Such an effective dose will generally depend upon the factors described herein. If desired, an effective daily dose of immune modulator can be administered as two, three, four, five, or six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.

[1707] In some examples, administration of an immune modulator using any of the compositions or devices described herein can result in the onset of treatment (e.g., a reduction in the number, severity, or duration of one or more symptoms and/or markers of any of the inflammatory diseases or conditions that arise in tissue originating from the endoderm described herein) or drug-target engagement in a subject within a time period of about 10 minutes to about 10 hours, about 10 minutes to about 9 hours, about 10 minutes to about 8 hours, about 10 minutes to about 7 hours, about 10 minutes to about 6 hours, about 10 minutes to about 5 hours, about 10 minutes to about 4.5 hours, about 10 minutes to about 4 hours, about 10 minutes to about 3.5 hours, about 10 minutes to about 3 hours, about 10 minutes to about 2.5 hours, about 10 minutes to about 2 hours, about 10 minutes to about 1.5 hours, about 10 minutes to about 1 hour, about 10 minutes to about 55 minutes, about 10 minutes to about 50 minutes, about 10 minutes to about 45 minutes, about 10 minutes to about 40 minutes, about 10 minutes to about 35 minutes, about 10 minutes to about 30 minutes, about 10 minutes to about 25 minutes, about 10 minutes to about 20 minutes, about 10 minutes to about 15 minutes, about 15 minutes to about 10 hours, about 15 minutes to about 9 hours, about 15 minutes to about 8 hours, about 15 minutes to about 7 hours, about 15 minutes to about 6 hours, about 15 minutes to about 5 hours, about 15 minutes to about 4.5 hours, about 15 minutes to about 4 hours, about 15 minutes to about 3.5 hours, about 15 minutes to about 3 hours, about 15 minutes to about 2.5 hours, about 15 minutes to about 2 hours, about 15 minutes to about 1.5 hours, about 15 minutes to about 1 hour, about 15 minutes to about 55 minutes, about 15 minutes to about 50 minutes, about 15 minutes to about 45 minutes, about 15 minutes to about 40 minutes, about 15 minutes to about 35 minutes, about 15 minutes to about 30 minutes, about 15 minutes to about 25 minutes, about 15 minutes to about 20 minutes, about 20 minutes to about 10 hours, about 20 minutes to about 9 hours, about 20 minutes to about 8 hours, about 20 minutes to about 7 hours, about 20 minutes to about 6 hours, about 20 minutes to about 5 hours, about 20 minutes to about 4.5 hours, about 20 minutes to about 4 hours, about 20 minutes to about 3.5 hours, about 20 minutes to about 3 hours, about 20 minutes to about 2.5 hours, about 20 minutes to about 2 hours, about 20 minutes to about 1.5 hours, about 20 minutes to about 1 hour, about 20 minutes to about 55 minutes, about 20 minutes to about 50 minutes, about 20 minutes to about 45 minutes, about 20 minutes to about 40 minutes, about 20 minutes to about 35 minutes, about 20 minutes to about 30 minutes, about 20 minutes to about 25 minutes, about 25 minutes to about 10 hours, about 25 minutes to about 9 hours, about 25 minutes to about 8 hours, about 25 minutes to about 7 hours, about 25 minutes to about 6 hours, about 25 minutes to about 5 hours, about 25 minutes to about 4.5 hours, about 25 minutes to about 4 hours, about 25 minutes to about 3.5 hours, about 25 minutes to about 3 hours, about 25 minutes to about 2.5 hours, about 25 minutes to about 2 hours, about 25 minutes to about 1.5 hours, about 25 minutes to about 1 hour, about 25 minutes to about 55 minutes, about 25 minutes to about 50 minutes, about 25 minutes to about 45 minutes, about 25 minutes to about 40 minutes, about 25 minutes to about 35 minutes, about 25 minutes to about 30 minutes, about 30 minutes to about 10 hours, about 30 minutes to about 9 hours, about 30 minutes to about 8 hours, about 30 minutes to about 7 hours, about 30 minutes to about 6 hours, about 30 minutes to about 5 hours, about 30 minutes to about 4.5 hours, about 30 minutes to about 4 hours, about 30 minutes to about 3.5 hours, about 30 minutes to about 3 hours, about 30 minutes to about 2.5 hours, about 30 minutes to about 2 hours, about 30 minutes to about 1.5 hours, about 30 minutes to about 1 hour, about 30 minutes to about 55 minutes, about 30 minutes to about 50 minutes, about 30 minutes to about 45 minutes, about 30 minutes to about 40 minutes, about 30 minutes to about 35 minutes, about 35 minutes to about 10 hours, about 35 minutes to about 9 hours, about 35 minutes to about 8 hours, about 35 minutes to about 7 hours, about 35 minutes to about 6 hours, about 35 minutes to about 5 hours, about 35 minutes to about 4.5 hours, about 35 minutes to about 4 hours, about 35 minutes to about 3.5 hours, about 35 minutes to about 3 hours, about 35 minutes to about 2.5 hours, about 35 minutes to about 2 hours, about 35 minutes to about 1.5 hours, about 35 minutes to about 1 hour, about 35 minutes to about 55 minutes, about 35 minutes to about 50 minutes, about 35 minutes to about 45 minutes, about 35 minutes to about 40 minutes, about 40 minutes to about 10 hours, about 40 minutes to about 9 hours, about 40 minutes to about 8 hours, about 40 minutes to about 7 hours, about 40 minutes to about 6 hours, about 40 minutes to about 5 hours, about 40 minutes to about 4.5 hours, about 40 minutes to about 4 hours, about 40 minutes to about 3.5 hours, about 40 minutes to about 3 hours, about 40 minutes to about 2.5 hours, about 40 minutes to about 2 hours, about 40 minutes to about 1.5 hours, about 40 minutes to about 1 hour, about 40 minutes to about 55 minutes, about 40 minutes to about 50 minutes, about 40 minutes to about 45 minutes, about 45 minutes to about 10 hours, about 45 minutes to about 9 hours, about 45 minutes to about 8 hours, about 45 minutes to about 7 hours, about 45 minutes to about 6 hours, about 45 minutes to about 5 hours, about 45 minutes to about 4.5 hours, about 45 minutes to about 4 hours, about 45 minutes to about 3.5 hours, about 45 minutes to about 3 hours, about 45 minutes to about 2.5 hours, about 45 minutes to about 2 hours, about 45 minutes to about 1.5 hours, about 45 minutes to about 1 hour, about 45 minutes to about 55 minutes, about 45 minutes to about 50 minutes, about 50 minutes to about 10 hours, about 50 minutes to about 9 hours, about 50 minutes to about 8 hours, about 50 minutes to about 7 hours, about 50 minutes to about 6 hours, about 50 minutes to about 5 hours, about 50 minutes to about 4.5 hours, about 50 minutes to about 4 hours, about 50 minutes to about 3.5 hours, about 50 minutes to about 3 hours, about 50 minutes to about 2.5 hours, about 50 minutes to about 2 hours, about 50 minutes to about 1.5 hours, about 50 minutes to about 1 hour, about 50 minutes to about 55 minutes, about 55 minutes to about 10 hours, about 55 minutes to about 9 hours, about 55 minutes to about 8 hours, about 55 minutes to about 7 hours, about 55 minutes to about 6 hours, about 55 minutes to about 5 hours, about 55 minutes to about 4.5 hours, about 55 minutes to about 4 hours, about 55 minutes to about 3.5 hours, about 55 minutes to about 3 hours, about 55 minutes to about 2.5 hours, about 55 minutes to about 2 hours, about 55 minutes to about 1.5 hours, about 55 minutes to about 1 hour, about 1 hour to about 10 hours, about 1 hour to about 9 hours, about 1 hour to about 8 hours, about 1 hour to about 7 hours, about 1 hour to about 6 hours, about 1 hour to about 5 hours, about 1 hour to about 4.5 hours, about 1 hour to about 4 hours, about 1 hour to about 3.5 hours, about 1 hour to about 3 hours, about 1 hour to about 2.5 hours, about 1 hour to about 2 hours, about 1 hour to about 1.5 hours, about 1.5 hours to about 10 hours, about 1.5 hours to about 9 hours, about 1.5 hours to about 8 hours, about 1.5 hours to about 7 hours, about 1.5 hours to about 6 hours, about 1.5 hours to about 5 hours, about 1.5 hours to about 4.5 hours, about 1.5 hours to about 4 hours, about 1.5 hours to about 3.5 hours, about 1.5 hours to about 3 hours, about 1.5 hours to about 2.5 hours, about 1.5 hours to about 2 hours, about 2 hours to about 10 hours, about 2 hours to about 9 hours, about 2 hours to about 8 hours, about 2 hours to about 7 hours, about 2 hours to about 6 hours, about 2 hours to about 5 hours, about 2 hours to about 4.5 hours, about 2 hours to about 4 hours, about 2 hours to about 3.5 hours, about 2 hours to about 3 hours, about 2 hours to about 2.5 hours, about 2.5 hours to about 10 hours, about 2.5 hours to about 9 hours, about 2.5 hours to about 8 hours, about 2.5 hours to about 7 hours, about 2.5 hours to about 6 hours, about 2.5 hours to about 5 hours, about 2.5 hours to about 4.5 hours, about 2.5 hours to about 4 hours, about 2.5 hours to about 3.5 hours, about 2.5 hours to about 3 hours, about 3 hours to about 10 hours, about 3 hours to about 9 hours, about 3 hours to about 8 hours, about 3 hours to about 7 hours, about 3 hours to about 6 hours, about 3 hours to about 5 hours, about 3 hours to about 4.5 hours, about 3 hours to about 4 hours, about 3 hours to about 3.5 hours, about 3.5 hours to about 10 hours, about 3.5 hours to about 9 hours, about 3.5 hours to about 8 hours, about 3.5 hours to about 7 hours, about 3.5 hours to about 6 hours, about 3.5 hours to about 5 hours, about 3.5 hours to about 4.5 hours, about 3.5 hours to about 4 hours, about 4 hours to about 10 hours, about 4 hours to about 9 hours, about 4 hours to about 8 hours, about 4 hours to about 7 hours, about 4 hours to about 6 hours, about 4 hours to about 5 hours, about 4 hours to about 4.5 hours, about 4.5 hours to about 10 hours, about 4.5 hours to about 9 hours, about 4.5 hours to about 8 hours, about 4.5 hours to about 7 hours, about 4.5 hours to about 6 hours, about 4.5 hours to about 5 hours, about 5 hours to about 10 hours, about 5 hours to about 9 hours, about 5 hours to about 8 hours, about 5 hours to about 7 hours, about 5 hours to about 6 hours, about 6 hours to about 10 hours, about 6 hours to about 9 hours, about 6 hours to about 8 hours, about 6 hours to about 7 hours, about 7 hours to about 10 hours, about 7 hours to about 9 hours, about 7 hours to about 8 hours, about 8 hours to about 10 hours, about 8 hours to about 9 hours, or about 9 hours to about 10 hours of administration of a dose of an immune modulator using any of the devices or compositions described herein. Drug-target engagement may be determined, for example, as disclosed in Simon G M, Niphakis M J, Cravatt B F, Nature chemical biology. 2013; 9(4):200-205, incorporated by reference herein in its entirety.

[1708] In some embodiments, administration of an immune modulator using any of the devices or compositions described herein can provide for treatment (e.g., a reduction in the number, severity, and/or duration of one or more symptoms and/or markers of any of the inflammatory diseases or conditions that arise in a tissue originating from the endoderm in a subject) for a time period of between about 1 hour to about 30 days, about 1 hour to about 28 days, about 1 hour to about 26 days, about 1 hour to about 24 days, about 1 hour to about 22 days, about 1 hour to about 20 days, about 1 hour to about 18 days, about 1 hour to about 16 days, about 1 hour to about 14 days, about 1 hour to about 12 days, about 1 hour to about 10 days, about 1 hour to about 8 days, about 1 hour to about 6 days, about 1 hour to about 5 days, about 1 hour to about 4 days, about 1 hour to about 3 days, about 1 hour to about 2 days, about 1 hour to about 1 day, about 1 hour to about 12 hours, about 1 hour to about 6 hours, about 1 hour to about 3 hours, about 3 hours to about 30 days, about 3 hours to about 28 days, about 3 hours to about 26 days, about 3 hours to about 24 days, about 3 hours to about 22 days, about 3 hours to about 20 days, about 3 hours to about 18 days, about 3 hours to about 16 days, about 3 hours to about 14 days, about 3 hours to about 12 days, about 3 hours to about 10 days, about 3 hours to about 8 days, about 3 hours to about 6 days, about 3 hours to about 5 days, about 3 hours to about 4 days, about 3 hours to about 3 days, about 3 hours to about 2 days, about 3 hours to about 1 day, about 3 hours to about 12 hours, about 3 hours to about 6 hours, about 6 hours to about 30 days, about 6 hours to about 28 days, about 6 hours to about 26 days, about 6 hours to about 24 days, about 6 hours to about 22 days, about 6 hours to about 20 days, about 6 hours to about 18 days, about 6 hours to about 16 days, about 6 hours to about 14 days, about 6 hours to about 12 days, about 6 hours to about 10 days, about 6 hours to about 8 days, about 6 hours to about 6 days, about 6 hours to about 5 days, about 6 hours to about 4 days, about 6 hours to about 3 days, about 6 hours to about 2 days, about 6 hours to about 1 day, about 6 hours to about 12 hours, about 12 hours to about 30 days, about 12 hours to about 28 days, about 12 hours to about 26 days, about 12 hours to about 24 days, about 12 hours to about 22 days, about 12 hours to about 20 days, about 12 hours to about 18 days, about 12 hours to about 16 days, about 12 hours to about 14 days, about 12 hours to about 12 days, about 12 hours to about 10 days, about 12 hours to about 8 days, about 12 hours to about 6 days, about 12 hours to about 5 days, about 12 hours to about 4 days, about 12 hours to about 3 days, about 12 hours to about 2 days, about 12 hours to about 1 day, about 1 day to about 30 days, about 1 day to about 28 days, about 1 day to about 26 days, about 1 day to about 24 days, about 1 day to about 22 days, about 1 day to about 20 days, about 1 day to about 18 days, about 1 day to about 16 days, about 1 day to about 14 days, about 1 day to about 12 days, about 1 day to about 10 days, about 1 day to about 8 days, about 1 day to about 6 days, about 1 day to about 5 days, about 1 day to about 4 days, about 1 day to about 3 days, about 1 day to about 2 days, about 2 days to about 30 days, about 2 days to about 28 days, about 2 days to about 26 days, about 2 days to about 24 days, about 2 days to about 22 days, about 2 days to about 20 days, about 2 days to about 18 days, about 2 days to about 16 days, about 2 days to about 14 days, about 2 days to about 12 days, about 2 days to about 10 days, about 2 days to about 8 days, about 2 days to about 6 days, about 2 days to about 5 days, about 2 days to about 4 days, about 2 days to about 3 days, about 3 days to about 30 days, about 3 days to about 28 days, about 3 days to about 26 days, about 3 days to about 24 days, about 3 days to about 22 days, about 3 days to about 20 days, about 3 days to about 18 days, about 3 days to about 16 days, about 3 days to about 14 days, about 3 days to about 12 days, about 3 days to about 10 days, about 3 days to about 8 days, about 3 days to about 6 days, about 3 days to about 5 days, about 3 days to about 4 days, about 4 days to about 30 days, about 4 days to about 28 days, about 4 days to about 26 days, about 4 days to about 24 days, about 4 days to about 22 days, about 4 days to about 20 days, about 4 days to about 18 days, about 4 days to about 16 days, about 4 days to about 14 days, about 4 days to about 12 days, about 4 days to about 10 days, about 4 days to about 8 days, about 4 days to about 6 days, about 4 days to about 5 days, about 5 days to about 30 days, about 5 days to about 28 days, about 5 days to about 26 days, about 5 days to about 24 days, about 5 days to about 22 days, about 5 days to about 20 days, about 5 days to about 18 days, about 5 days to about 16 days, about 5 days to about 14 days, about 5 days to about 12 days, about 5 days to about 10 days, about 5 days to about 8 days, about 5 days to about 6 days, about 6 days to about 30 days, about 6 days to about 28 days, about 6 days to about 26 days, about 6 days to about 24 days, about 6 days to about 22 days, about 6 days to about 20 days, about 6 days to about 18 days, about 6 days to about 16 days, about 6 days to about 14 days, about 6 days to about 12 days, about 6 days to about 10 days, about 6 days to about 8 days, about 8 days to about 30 days, about 8 days to about 28 days, about 8 days to about 26 days, about 8 days to about 24 days, about 8 days to about 22 days, about 8 days to about 20 days, about 8 days to about 18 days, about 8 days to about 16 days, about 8 days to about 14 days, about 8 days to about 12 days, about 8 days to about 10 days, about 10 days to about 30 days, about 10 days to about 28 days, about 10 days to about 26 days, about 10 days to about 24 days, about 10 days to about 22 days, about 10 days to about 20 days, about 10 days to about 18 days, about 10 days to about 16 days, about 10 days to about 14 days, about 10 days to about 12 days, about 12 days to about 30 days, about 12 days to about 28 days, about 12 days to about 26 days, about 12 days to about 24 days, about 12 days to about 22 days, about 12 days to about 20 days, about 12 days to about 18 days, about 12 days to about 16 days, about 12 days to about 14 days, about 14 days to about 30 days, about 14 days to about 28 days, about 14 days to about 26 days, about 14 days to about 24 days, about 14 days to about 22 days, about 14 days to about 20 days, about 14 days to about 18 days, about 14 days to about 16 days, about 16 days to about 30 days, about 16 days to about 28 days, about 16 days to about 26 days, about 16 days to about 24 days, about 16 days to about 22 days, about 16 days to about 20 days, about 16 days to about 18 days, about 18 days to about 30 days, about 18 days to about 28 days, about 18 days to about 26 days, about 18 days to about 24 days, about 18 days to about 22 days, about 18 days to about 20 days, about 20 days to about 30 days, about 20 days to about 28 days, about 20 days to about 26 days, about 20 days to about 24 days, about 20 days to about 22 days, about 22 days to about 30 days, about 22 days to about 28 days, about 22 days to about 26 days, about 22 days to about 24 days, about 24 days to about 30 days, about 24 days to about 28 days, about 24 days to about 26 days, about 26 days to about 30 days, about 26 days to about 28 days, or about 28 days to about 30 days in a subject following first administration of an immune modulator using any of the compositions or devices described herein. Non-limiting examples of symptoms and/or markers of a disease described herein are described below.

[1709] For example, treatment can result in a decrease (e.g., about 1% to about 99% decrease, about 1% to about 95% decrease, about 1% to about 90% decrease, about 1% to about 85% decrease, about 1% to about 80% decrease, about 1% to about 75% decrease, about 1% to about 70% decrease, about 1% to about 65% decrease, about 1% to about 60% decrease, about 1% to about 55% decrease, about 1% to about 50% decrease, about 1% to about 45% decrease, about 1% to about 40% decrease, about 1% to about 35% decrease, about 1% to about 30% decrease, about 1% to about 25% decrease, about 1% to about 20% decrease, about 1% to about 15% decrease, about 1% to about 10% decrease, about 1% to about 5% decrease, about 5% to about 99% decrease, about 5% to about 95% decrease, about 5% to about 90% decrease, about 5% to about 85% decrease, about 5% to about 80% decrease, about 5% to about 75% decrease, about 5% to about 70% decrease, about 5% to about 65% decrease, about 5% to about 60% decrease, about 5% to about 55% decrease, about 5% to about 50% decrease, about 5% to about 45% decrease, about 5% to about 40% decrease, about 5% to about 35% decrease, about 5% to about 30% decrease, about 5% to about 25% decrease, about 5% to about 20% decrease, about 5% to about 15% decrease, about 5% to about 10% decrease, about 10% to about 99% decrease, about 10% to about 95% decrease, about 10% to about 90% decrease, about 10% to about 85% decrease, about 10% to about 80% decrease, about 10% to about 75% decrease, about 10% to about 70% decrease, about 10% to about 65% decrease, about 10% to about 60% decrease, about 10% to about 55% decrease, about 10% to about 50% decrease, about 10% to about 45% decrease, about 10% to about 40% decrease, about 10% to about 35% decrease, about 10% to about 30% decrease, about 10% to about 25% decrease, about 10% to about 20% decrease, about 10% to about 15% decrease, about 15% to about 99% decrease, about 15% to about 95% decrease, about 15% to about 90% decrease, about 15% to about 85% decrease, about 15% to about 80% decrease, about 15% to about 75% decrease, about 15% to about 70% decrease, about 15% to about 65% decrease, about 15% to about 60% decrease, about 15% to about 55% decrease, about 15% to about 50% decrease, about 15% to about 45% decrease, about 15% to about 40% decrease, about 15% to about 35% decrease, about 15% to about 30% decrease, about 15% to about 25% decrease, about 15% to about 20% decrease, about 20% to about 99% decrease, about 20% to about 95% decrease, about 20% to about 90% decrease, about 20% to about 85% decrease, about 20% to about 80% decrease, about 20% to about 75% decrease, about 20% to about 70% decrease, about 20% to about 65% decrease, about 20% to about 60% decrease, about 20% to about 55% decrease, about 20% to about 50% decrease, about 20% to about 45% decrease, about 20% to about 40% decrease, about 20% to about 35% decrease, about 20% to about 30% decrease, about 20% to about 25% decrease, about 25% to about 99% decrease, about 25% to about 95% decrease, about 25% to about 90% decrease, about 25% to about 85% decrease, about 25% to about 80% decrease, about 25% to about 75% decrease, about 25% to about 70% decrease, about 25% to about 65% decrease, about 25% to about 60% decrease, about 25% to about 55% decrease, about 25% to about 50% decrease, about 25% to about 45% decrease, about 25% to about 40% decrease, about 25% to about 35% decrease, about 25% to about 30% decrease, about 30% to about 99% decrease, about 30% to about 95% decrease, about 30% to about 90% decrease, about 30% to about 85% decrease, about 30% to about 80% decrease, about 30% to about 75% decrease, about 30% to about 70% decrease, about 30% to about 65% decrease, about 30% to about 60% decrease, about 30% to about 55% decrease, about 30% to about 50% decrease, about 30% to about 45% decrease, about 30% to about 40% decrease, about 30% to about 35% decrease, about 35% to about 99% decrease, about 35% to about 95% decrease, about 35% to about 90% decrease, about 35% to about 85% decrease, about 35% to about 80% decrease, about 35% to about 75% decrease, about 35% to about 70% decrease, about 35% to about 65% decrease, about 35% to about 60% decrease, about 35% to about 55% decrease, about 35% to about 50% decrease, about 35% to about 45% decrease, about 35% to about 40% decrease, about 40% to about 99% decrease, about 40% to about 95% decrease, about 40% to about 90% decrease, about 40% to about 85% decrease, about 40% to about 80% decrease, about 40% to about 75% decrease, about 40% to about 70% decrease, about 40% to about 65% decrease, about 40% to about 60% decrease, about 40% to about 55% decrease, about 40% to about 50% decrease, about 40% to about 45% decrease, about 45% to about 99% decrease, about 45% to about 95% decrease, about 45% to about 90% decrease, about 45% to about 85% decrease, about 45% to about 80% decrease, about 45% to about 75% decrease, about 45% to about 70% decrease, about 45% to about 65% decrease, about 45% to about 60% decrease, about 45% to about 55% decrease, about 45% to about 50% decrease, about 50% to about 99% decrease, about 50% to about 95% decrease, about 50% to about 90% decrease, about 50% to about 85% decrease, about 50% to about 80% decrease, about 50% to about 75% decrease, about 50% to about 70% decrease, about 50% to about 65% decrease, about 50% to about 60% decrease, about 50% to about 55% decrease, about 55% to about 99% decrease, about 55% to about 95% decrease, about 55% to about 90% decrease, about 55% to about 85% decrease, about 55% to about 80% decrease, about 55% to about 75% decrease, about 55% to about 70% decrease, about 55% to about 65% decrease, about 55% to about 60% decrease, about 60% to about 99% decrease, about 60% to about 95% decrease, about 60% to about 90% decrease, about 60% to about 85% decrease, about 60% to about 80% decrease, about 60% to about 75% decrease, about 60% to about 70% decrease, about 60% to about 65% decrease, about 65% to about 99% decrease, about 65% to about 95% decrease, about 65% to about 90% decrease, about 65% to about 85% decrease, about 65% to about 80% decrease, about 65% to about 75% decrease, about 65% to about 70% decrease, about 70% to about 99% decrease, about 70% to about 95% decrease, about 70% to about 90% decrease, about 70% to about 85% decrease, about 70% to about 80% decrease, about 70% to about 75% decrease, about 75% to about 99% decrease, about 75% to about 95% decrease, about 75% to about 90% decrease, about 75% to about 85% decrease, about 75% to about 80% decrease, about 80% to about 99% decrease, about 80% to about 95% decrease, about 80% to about 90% decrease, about 80% to about 85% decrease, about 85% to about 99% decrease, about 85% to about 95% decrease, about 85% to about 90% decrease, about 90% to about 99% decrease, about 90% to about 95% decrease, or about 95% to about 99% decrease) in one or more (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve) of the severity of one or more symptoms of the inflammatory disease or condition that arises in a tissue originating from the endoderm, a decrease in the number of memory Th cells present in a meserteric lymph node, a decrease in the expression of 47 integrin in memory Th cells present in a mesenteric lymph node, a decrease in the number of memory Th cells present in the Peyer's patch, and a decrease in the expression of 47 integrin in memory Th cells present in the Peyer's patch, a decrease in the level of interferon- in the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of IL-I Pin the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of IL-6 in the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of IL-22 in the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of IL-17A in the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of TNF in the tissue originating from the endoderm involved in the inflammatory disease or condition, a decrease in the level of IL-2 in the tissue originating from the endoderm involved in the inflammatory disease or condition, and a decrease in the number of T-lymphocytes that have migrated into the tissue originating from the endoderm involved in the inflammatory disease or condition, in a subject (e.g., as compared to the level in the subject prior to treatment or compared to a subject or population of subjects having a similar disease but receiving a placebo or a different treatment) (e.g., for a time period of between about 1 hour to about 30 days (e.g., or any of the subranges herein) following the first administration of an immune modulator using any of the compositions or devices described herein. As used herein, GI tissue refers to tissue in the gastrointestinal (GI) tract, such as tissue in one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum, more particularly in the proximal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon, or in the distal portion of one or more of duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and sigmoid colon. Exemplary methods for determining the levels of interferon-, IL-1p, IL-6, IL-22, IL-17A, TNF, and IL-2 are described herein. Additional methods for determining the levels of these cytokines are known in the art. Exemplary methods for determining the number of Th memory cells in Peyer's patches and mesentery lymph nodes are described herein. Additional methods for determining the number of Th memory cells in Peyer's patches and mesentery lymph nodes are known in the art.

[1710] Accordingly, in some embodiments, a method of treatment disclosed herein includes determining the level of a marker at the location of disease in a subject (e.g., either before and/or after administration of the device). In some embodiments, the marker is a biomarker and the method of treatment disclosed herein comprises determining that the level of a biomarker at the location of disease is a subject following administration of the device is decreased as compared to the level of the biomarker at the same location of disease in a subject either before administration or at the same time point following systemic administration of an equal amount of the immune modulator. In some examples, the level of the biomarker at the same location of disease following administration of the device is 1% decreased to 99% decreased as compared to the level of the biomarker at the same location of disease in a subject either before administration or at the same time point following systemic administration of an equal amount of the immune modulator. In some embodiments, the level of the marker is one or more of: the level of interferon-, the level of IL-17A, the level of TNF, the level of IL-2, the number of Th memory cells in Peyer's patches, and the number of Th memory cells in mesenteric lymph nodes.

[1711] In some embodiments, the method of treatment disclosed herein includes determining that the level of a marker at a time point following administration of a device is lower than the level of the marker at a time point following administration of the device is lower than the level of the marker in a subject prior to administration of the device or in a subject at substantially the same time point following systemic administration of an equal amount of the immune modulator. In some examples, the level of the marker following administration of the device is 1% decreased to 99% decreased as compared to the level of the marker in a subject prior to administration of the device or in a subject at the same time point following systemic administration of an equal amount of the immune modulator. In some examples, a method of treatment disclosed herein includes determining the level of the biomarker at the location of disease in a subject within a time period of about 10 minutes to 10 hours following administration of the device.

[1712] In some embodiments, a method of treatment described herein includes: (i) determining the ratio RB of the level LiB of a biomarker at the location of disease at a first time point following administration of the device and the level L2B of the biomarker at the same location of disease in a subject at substantially the same time point following systemic administration of an equal amount of the immune modulator; (ii) determining the ratio of R.sub.D of the level of L.sub.1D of the immune modulator at the same location and the substantially the same time point as in (i) and the level L.sub.2D of the immune modulator at the same location of disease in a subject at substantially the same time point following systemic administration of an equal amount of the immune modulator; and (iii) determining the ratio of R.sub.B/R.sub.D.

[1713] In some embodiments, a method of treatment disclosed herein can include: (i) determining the ratio R.sub.B of the level L.sub.1B of a biomarker at the location of disease at a time point following administration of the device and the level L2B of the biomarker at the same location of disease in a subject at substantially the same time point following systemic administration of an equal amount of the immune modulator; (ii) determining the ratio R.sub.D of the level L.sub.1D of the immune modulator at the same location and at substantially the time point as in (i) and the level L.sub.2D of the immune modulator in a subject at the same location of disease at substantially the same time point following systemic administration of an equal amount of the immune modulator; and (iii) determining the product R.sub.BR.sub.D.

[1714] In some embodiments, a method of treatment disclosed herein can include determining that the level of a marker in a subject at a time point following administration of the device is elevated as compared to a level of the marker in a subject prior to administration of the device or a level at substantially the same time point in a subject following systemic administration of an equal amount of the immune modulator. In some examples, the level of the marker at a time point following administration of the device is 1% increased or 400% increased as compared to the level of the marker in a subject prior to administration of the device or a level at substantially the same time point in a subject following systemic administration of an equal amount of the immune modulator. In some examples, a method of treatment disclosed herein includes determining the level of the marker in a subject within a period of about 10 minutes to about 10 hours following administration of the device.

[1715] In some embodiments, a method of treatment disclosed herein can include determining the level of a marker in a subject's blood, serum or plasma.

[1716] An illustrative list of examples of biomarkers for GI disorders includes interferon-, IL-1, IL-6, IL-22, IL-17A, TNF, IL-2, memory cells (CD44.sup.+CD45RB.sup.CD4.sup.+ cells); 47; VEGF; ICAM; VCAM; SAA; Calprotectin; lactoferrin; FGF2; TGFb; ANG-1; ANG-2; PLGF; Biologics (Infliximab; Humira; Stelara; Vedolizumab; Simponi; Jak inhibitors; Others); EGF; IL12/23p40; GMCSF; A4 B7; AeB7; CRP; SAA; ICAM; VCAM; AREG; EREG; HB-EGF; HRG; BTC; TGF; SCF; TWEAK; MMP-9; MMP-6; Ceacam CD66; IL10; ADA; Madcam-1; CD166 (AL CAM); FGF2; FGF7; FGF9; FGF19; ANCA Antineutrophil cytoplasmic antibody; ASCAA Anti-Saccharomyces Cerevisiae Antibody IgA; ASCAG Anti-Saccharomyces Cerevisiae Antibody IgG; CBir1 Anti-Clostridium cluster XIVa flagellin CBir1 antibody; A4-Fla2 Anti-Clostridium cluster XIVa flagellin 2 antibody; FlaX Anti-Clostridium cluster XIVa flagellin X antibody; OmpC Anti-Escherichia coli Outer Membrane Protein C; ANCA Perinuclear AntiNeutrophil Cytoplasmic Antibody; AREG Amphiregulin Protein; BTC Betacellulin Protein; EGF Epidermal Growth Factor EREG

[1717] Epiregulin Protein; HBEGF Heparin Binding Epidermal Growth Factors; HGF Hepatocyte Growth Factor; HRG Neuregulin-1; TGFA Transforming Growth Factor alpha; CRP C-Reactive Protein; SAA Serum Amyloid A; ICAM-1 Intercellular Adhesion Molecule 1; VCAM-1 Vascular Cell Adhesion Molecule 1; fibroblasts underlying the intestinal epithelium; and HGF.

[1718] In some embodiments, a marker is an IBD biomarker, such as, for example: anti-glycan; anti-Saccharomices cerevisiae (ASCA); anti-laminaribioside (ALCA); anti-chitobioside (ACCA); anti-mannobioside (AMCA); anti-laminarin (anti-L); anti-chitin (anti-C) antibodies: anti-outer membrane porin C (anti-OmpC), anti-Cbir1 flagellin; anti-12 antibody; autoantibodies targeting the exocrine pancreas (PAB); and perinuclear antineutrophil antibody (pANCA); and calprotectin.

[1719] In some embodiments, a biomarker is associated with membrane repair, fibrosis, angiogenesis. In certain embodiments, a biomarker is an inflammatory biomarker, an anti-inflammatory biomarker, an MMP biomarker, an immune marker, or a TNF pathway biomarker. In some embodiments, a biomarker is gut specific.

[1720] For tissue samples, HER2 can be used as a biomarker relating to cytotoxic T cells. Additionally, other cytokine levels can be used as biomarkers in tissue (e.g., phospho STAT 1, STAT 3 and STAT 5), in plasma (e.g., VEGF, VCAM, ICAM, IL-6), or both.

[1721] In some embodiments, the target analyte(s) include one or more immunoglobulins, such as, for example, immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin G (IgG), immunoglobulin E (IgE) and/or immunoglobulin A (IgA). In some embodiments, IgM is a biomarker of infection and/or inflammation. In some embodiments, IgD is a biomarker of autoimmune disease. In some embodiments, IgG is a biomarker of Alzheimer's disease and/or for cancer. In some embodiments, IgE is a biomarker of asthma and/or allergen immunotherapy. In some embodiments, IgA is a biomarker of kidney disease.

[1722] In some embodiments, a biomarker or marker of a liver disease or disorder (e.g., any of the liver diseases or disorders described herein) is a bile acid or a bile salt (also known as a conjugated bile acid). Bile acids are products of cholesterol synthesis that are synthesized in the liver, conjugated to taurine or glycine, and stored in the gallbladder until released into the small intestine. The primary bile acids are cholic acid, and chenodeoxycholic acid, which are deconjugated and dehydroxylated by instestinal bacteria to form the secondary bile acids deoxycholic acid and lithocholic acid, respectively. The majority of bile acids (about 95%) are reabsorbed in the distal ileum and returned to the liver (see, e.g., U.S. Publication No. 2017/0343535, incorporated herein by reference). Impaired absorption of bile acids in the ileum can lead to excess bile acids in the colon which can cause symptoms of bile acid malabsorption (BAM; also known as bile acid diarrhea), including watery stool and fecal incontinence. Interestingly, up to 50% of patients with irritable bowel syndrome with diarrhea (IBS-D) also have BAM (see, e.g., Camilleri et al. (2009) Neurogastroeterol. Motil. 21(7): 734-43). In some embodiments, the presence, absence, and/or a specific level of one or more bile acids or bile salts in the GI tract of a subject is indicative of a condition or disease state (e.g., a GI disorder and/or a non-GI disorder (e.g., a systemic disorder or a liver disease)). In some embodiments, the level of at least one bile acid or bile salt in the GI tract of the subject is used to diagnose a GI disorder such as BAM or IBS (e.g., IBS-D). In some embodiments, a level of a bile acid or a bile salt in the GI tract of a subject is determined. For instance, the presence and/or absence, and/or the concentration of a bile acid, a bile salt, or a combination thereof, may be determined at a specific region of the GI tract of a subject (e.g., one or more of the duodenum, jejunum, ileum, ascending colon, transverse colon or descending colon) to determine whether the subject has or is at risk of developing a GI disorder, such as BAM or IBS-D. In some embodiments, the ratio of two or more bile acids or bile acid salts in the GI tract of a subject (e.g., a specific region of the GI tract of a subject including one or more of the duodenum, jejunum, ileum, ascending colon, transverse colon or descending colon) can be determined. In some embodiments, the presence and/or absence, and/or the concentration of a bile acid, a bile salt, or a combination thereof, is determined in the ileum of a subject. In some embodiments, the presence and/or absence, and/or the concentration of a bile acid, a bile salt, or a combination thereof, is determined in the colon of a subject. In some embodiments, the concentration of a bile acid, a bile salt, or a combination thereof, is determined in specific regions of the GI tract of the subject, and for example, compared to determine where along the GI tract the compounds are accumulating. In some embodiments, the detection of a concentration of a bile acid, bile salt, or a combination thereof, in a specific region of the GI tract of the subject (e.g., the colon or the ileum) that is above a reference level of a bile acid, bile salt, or a combination thereof (e.g., the average level of a bile acid in healthy subjects) may be indicative of BAM and/or IBS-D in a subject. In some embodiments, the bile acid is selected from the group consisting of chenodeoxycholic acid, cholic acid, deoxycholate, lithocholate, and ursodeoxycholic acid. In some embodiments, the bile acid comprises cholesten-3-one or a structural variant thereof. In some embodiments, the bile acid is cholesten-3-one or a structural variant thereof. In some embodiments, the bile acid is cholesten-3-one. In some embodiments, the bile acid is a structural variant of cholesten-3-one. In some embodiments, the bile salt is selected from the group consisting of glycocholic acid, taurocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, glycolithocholic acid, and taurolithocholic acid.

[1723] Another biomarker of a liver disease or disorder is 7-hydroxy-4-cholesten-3-one (7C4). The measurement of 7C4 allows for the monitoring of the enzymatic activity of hepatic cholesterol 7-hydroxylase, the rate limiting enzyme in the synthesis of bile acids and can be used as a surrogate to detect BAM (see, e.g., Galman et al. (2003) J. Lipid. Res. 44: 859-66; and Camilleri et al. (2009) Neurogastroeterol. Motil. 21(7): 734-43, incorporated herein by reference in their entirety).

[1724] Biomarkers of a liver disease or disorder also include cholesterol, a lipid, a fat soluble vitamin (e.g., ascorbic acid, cholecalciferol, ergocalciferol, a tocopherol, a tocotrienol, phylloquinone, and a menaquinone), bilirubin, fibroblast growth factor 19 (FGF19), TGR5 (also known as GP-BAR1 or M-BAR), glycine, taurine, and cholecystokinin (CCK or CCK-PZ). In some embodiments, a biomarker of a liver disease or disorder is cholecystokinin. Cholecystokinin is a peptide hormone that contributes to control intestinal motility (see Rehfeld (2017) Front. Endocrinol. (Lausanne) 8: 47). In some embodiments, a biomarker of a liver disease or disorder is secretin. Secretin is a peptide hormone that regulates the pH of the duodenal content by controlling gastric acid secretion, regulates bile acid and bicarbonate secretion in the duodenum, and regulates water homeostasis (see, e.g., Afroze et al. (2013) Ann. Transl. Med. 1(3): 29). In some embodiments, a subject has previously been administered cholecystokinin or secretin to induce the release of a biomarker or marker (e.g., from the liver and/or gall bladder into the GI tract).

[1725] An illustrative list of examples of biomarkers that may be used to detect, diagnose, or monitor treatment efficacy for a liver disease or disorder include bilirubin, gamma-glutamyl transferase (GGT), haptoglobin, apolipoprotein A1, alpha2-macroglobulin, cholesterol, triglycerides, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose, cytokeratin-18 (CK18) fragment, hyaluronic acid, TGF-, fatty acid binding protein, hydroxysteroid 17-beta dehydrogenase 13 (17-HSD13), glutamyl dipeptides, glutamyl valine, glutamyl leucine, glutamyl phenylalanine, glutamyl tyrosine, carnitine, butylcarnitine, lysine, tyrosine, isoleucine, glycerophosphatidylcholine, glycerylphsphorylethanolamine, taurine, glycine conjugates, taurocholic acid, taurodeoxycholic acid, lactate, glutamate, cysteine-gluthatione disulfide, caprate, 10-undecenoate, oleoyl-lysophosphatidylcholine, oxidized and reduced gluthatione, glutamate, andenosine triphosphate, creatine, cholic acid, and glycodeoxycholic acid. In some embodiments, a biomarker of a liver disease or disorder can be a metabolite of any of the markers or biomarkers described herein.

[1726] In some embodiments, the biomarker is High Sensitivity C-reactive Protein (hsCRP); 7 -hydroxy-4-cholesten-3-one (7C4); Anti-Endomysial IgA (EMA IgA); Anti-Human Tissue Transglutaminase IgA (tTG IgA); Total Serum IgA by Nephelometry; Fecal Calprotectin; or Fecal Gastrointestinal Pathogens.

[1727] In some embodiments, the biomarker is

a) an anti-gliadin IgA antibody, an anti-gliadin IgG antibody, an anti-tissue transglutaminase (tTG) antibody, an anti-endomysial antibody;
b)i) a serological marker that is ASCA-A, ASCA-G, ANCA, pANCA, anti-OmpC antibody, anti-CBir1 antibody, anti-FlaX antibody, or anti-A4-Fla2 antibody;
b)ii) an inflammation marker that is VEGF, ICAM, VCAM, SAA, or CRP;
b)iii) the genotype of the genetic markers ATG16L1, ECM1, NKX2-3, or STAT3;
c) a bacterial antigen antibody marker;
d) a mast cell marker;
e) an inflammatory cell marker;
f) a bile acid malabsorption (BAM) marker;
g) a kynurenine marker;
or
h) a serotonin marker.

[1728] In some embodiments, the bacterial antigen antibody marker is selected from the group consisting of an anti-Fla1 antibody, anti-Fla2 antibody, anti-FlaA antibody, anti-FliC antibody, anti-FliC2 antibody, anti-FliC3 antibody, anti-YBaN1 antibody, anti-ECFliC antibody, anti-Ec0FliC antibody, anti-SeFljB antibody, anti-CjFlaA antibody, anti-CjFaB antibody, anti-SfFliC antibody, anti-CjCgtA antibody, anti-Cjdmh antibody, anti-CjGT-A antibody, anti-EcYidX antibody, anti-EcEra antibody, anti-EcFrvX antibody, anti-EcGabT antibody, anti-EcYedK antibody, anti-EcYbaN antibody, anti-EcYhgN antibody, anti-RtMaga antibody, anti-RbCpaF antibody, anti-RgPilD antibody, anti-LaFrc antibody, anti-LaEno antibody, anti-LjEFTu antibody, anti-BfOmpa antibody, anti-PrOmpA antibody, anti-Cp10bA antibody, anti-CpSpA antibody, anti-EfSant antibody, anti-LmOsp antibody, anti-SfET-2 antibody, anti-Cpatox antibody, anti-Cpbtox antibody, anti-EcSta2 antibody, anti-EcOStx2A antibody, anti-CjcdtB/C antibody, anti-CdtcdA/B antibody, and combinations thereof.

[1729] In some embodiments, the mast cell marker is selected from the group consisting of beta-tryptase, histamine, prostaglandin E2 (PGE2), and combinations thereof.

[1730] In some embodiments, the inflammatory marker is selected from the group consisting of CRP, ICAM, VCAM, SAA, GRO.alpha., and combinations thereof.

[1731] In some embodiments, the bile acid malabsorption marker is selected from the group consisting of 7-hydroxy-4-cholesten-3-one, FGF19, and a combination thereof.

[1732] In some embodiments, the kynurenine marker is selected from the group consisting of kynurenine (K), kynurenic acid (KyA), anthranilic acid (AA), 3-hydroxykynurenine (3-HK), 3-hydroxyanthranilic acid (3-HAA), xanthurenic acid (XA), quinolinic acid (QA), tryptophan, 5-hydroxytryptophan (5-HTP), and combinations thereof.

[1733] In some embodiments, the serotonin marker is selected from the group consisting of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), serotonin-O-sulfate, serotonin-O-phosphate, and combinations thereof.

[1734] In some embodiments, the biomarker is a biomarker as disclosed in U.S. Pat. No. 9,739,786, incorporated by reference herein in its entirety.

[1735] The following markers can be expressed by mesenchymal stem cells (MSC): CD105, CD73, CD90, CD13, CD29, CD44, CD10, Stro-1, CD271, SSEA-4, CD146, CD49f, CD349, GD2, 3G5, SSEA-3, SISD2, Stro-4, MSCA-1, CD56, CD200, PODX1, Sox1l, or TM4SF1 (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more of such markers), and lack expression of one or more of CD45, CD34, CD14, CD19, and HLA-DR (e.g., lack expression of two or more, three or more, four or more, or five or more such markers). In some embodiments, MSC can express CD105, CD73, and CD90. In some embodiments, MSC can express CD105, CD73, CD90, CD13, CD29, CD44, and CD10. In some embodiments, MSC can express CD105, CD73, and CD90 and one or more stemness markers such as Stro-1, CD271, SSEA-4, CD146, CD49f, CD349, GD2, 3G5, SSEA-3. SISD2, Stro-4, MSCA-1, CD56, CD200, PODX1, Sox11, or TM4SF1. In some embodiments, MSC can express CD105, CD73, CD90, CD13, CD29, CD44, and CD10 and one or more stemness markers such as Stro-1, CD271, SSEA-4, CD146, CD49f, CD349, GD2, 3G5, SSEA-3. SISD2, Stro-4, MSCA-1, CD56, CD200, PODX1, Sox1l, or TM4SF1. See, e.g., Lv, et al., Stem Cells, 2014, 32:1408-1419.

[1736] Intestinal stem cells (ISC) can be positive for one or more markers such as Musashi-1 (Msi-1), Ascl2, Bmi-1, Doublecortin and Ca2+/calmodulin-dependent kinase-like 1 (DCAMKL1), and Leucin-rich repeat-containing G-protein-coupled receptor 5 (Lgr5). See, e.g., Mohamed, et al., Cytotechnology, 2015 67(2): 177-189.

[1737] Any of the foregoing biomarkers can be used as a biomarker for one or more of other conditions as appropriate.

[1738] In some embodiments of the methods herein, the methods comprise determining the time period of onset of treatment following administration of the device.

Combination Therapy

[1739] The anti-inflamatory agents disclosed herein may be optionally be used with additional agents in the treatment of the diseases disclosed herein. Nonlimiting examples of such agents for treating or preventing inflammatory bowel disease in such adjunct therapy (e.g., Crohn's disease, ulcerative colitis) include substances that suppress cytokine production, down-regulate or suppress self-antigen expression, or mask the MHC antigens. Examples of such agents include 2-amino-6-aryl-5-substituted pyrimidines (see U.S. Pat. No. 4,665,077); non-steroidal antiinflammatory drugs (NSAIDs); ganciclovir; tacrolimus; lucocorticoids such as Cortisol or aldosterone; immune modulators such as a cyclooxygenase inhibitor; a 5-lipoxygenase inhibitor; or a leukotriene receptor antagonist; purine antagonists such as azathioprine or mycophenolate mofetil (MMF); alkylating agents such as cyclophosphamide; bromocryptine; danazol; dapsone; glutaraldehyde (which masks the MHC antigens, as described in U.S. Pat. No. 4,120,649); anti-idiotypic antibodies for MHC antigens and MHC fragments; cyclosporine; 6-mercaptopurine; steroids such as corticosteroids or glucocorticosteroids or glucocorticoid analogs, e.g., prednisone, methylprednisolone, including SOLU-MEDROL, methylprednisolone sodium succinate, and dexamethasone; dihydrofolate reductase inhibitors such as methotrexate (oral or subcutaneous); anti-malarial agents such as chloroquine and hydroxychloroquine; sulfasalazine; leflunomide; cytokine or cytokine receptor antibodies or antagonists including anti-interferon-alpha, -beta, or -gamma antibodies, anti-tumor necrosis factor(TNF)-alpha antibodies (infliximab (REMICADE) or adalimumab), anti-TNF-alpha immunoadhesin (etanercept), anti-TNF-beta antibodies, anti-interleukin-2 (IL-2) antibodies and anti-IL-2 receptor antibodies, and anti-interleukin-6 (IL-6) receptor antibodies and antagonists; anti-LFA-1 antibodies, including anti-CD1 la and anti-CD 18 antibodies; anti-L3T4 antibodies; heterologous anti-lymphocyte globulin; pan-T antibodies, anti-CD3 or anti-CD4/CD4a antibodies; soluble peptide containing a LFA-3 binding domain (WO 90/08187 published Jul. 26, 1990); streptokinase; transforming growth factor-beta (TGF-beta); streptodomase; RNA or DNA from the host; FK506; RS-61443; chlorambucil; deoxyspergualin; rapamycin; T-cell receptor (Cohen et al, U.S. Pat. No. 5,114,721); T-cell receptor fragments (Offner et al, Science, 251: 430-432 (1991); WO 90/11294; Ianeway, Nature, 341: 482 (1989); and WO 91/01133); BAFF antagonists such as BAFF or BR3 antibodies or immunoadhesins and zTNF4 antagonists (for review, see Mackay and Mackay, Trends Immunol, 23: 113-5 (2002) and see also definition below); biologic agents that interfere with T cell helper signals, such as anti-CD40 receptor or anti-CD40 ligand (CD 154), including blocking antibodies to CD40-CD40 ligand. (e.g., Durie et al, Science, 261: 1328-30 (1993); Mohan et al, J. Immunol, 154: 1470-80 (1995)) and CTLA4-Ig (Finck et al, Science, 265: 1225-7 (1994)); and T-cell receptor antibodies (EP 340,109) such as T10B9. Non-limiting examples of adjunct agents also include the following: budenoside; epidermal growth factor; aminosalicylates; metronidazole; mesalamine; olsalazine; balsalazide; antioxidants; thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1 monoclonal antibodies; growth factors; elastase inhibitors; pyridinyl-imidazole compounds; TNF antagonists; IL-4, IL-1, IL-13 and/or TGF cytokines or agonists thereof (e.g., agonist antibodies); IL-11; glucuronide- or dextran-conjugated prodrugs of prednisolone, dexamethasone or budesonide; ICAM-I antisense phosphorothioate oligodeoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TPlO; T Cell Sciences, Inc.); slow-release mesalazine; antagonists of platelet activating factor (PAF); ciprofloxacin; and lignocaine.

[1740] In other embodiments, an immune modulator as described herein can be administered with one or more of: an IL-12/IL-23 inhibitor, a CHST15 inhibitor, a IL-6 receptor inhibitor, a TNF inhibitor, an integrin inhibitor, a JAK inhibitor, a SMAD7 inhibitor, a IL-13 inhibitor, an IL-1 receptor inhibitor, a TLR agonist, an immunosuppressant, or a stem cell. In other embodiments, an immune modulator as described herein can be administered with a vitamin C infusion, one or more corticosteroids, and optionally thiamine.

Examples of particular combinations include the following. Unless otherwise specified, the first component (component (1)) is administered in an ingestible device, while the second component (component (2)) is administered either in an ingestible device, which may be the same or different ingestible device as the first component, or by another form of administration.
(1) Adalimumab; (2) methotrexate.
(1) Adalimumab; (2) methotrexate administered orally.
(1) Vedolizumab; (2) methotrexate.
(1) Vedolizumab; (2) methotrexate administered orally.
(1) Tacrolimus; (2) vedolizumab.
(1) Tacrolimus; (2) vedolizumab in an ingestible device.
(1) Tacrolimus; (2) vedolizumab intravenously or subcutaneously.
(1) A4 inhibitor; (2) Vedolizumab. In some embodiments, the A4 inhibitor is Tysabri.
(1) A4 inhibitor; (2) Vedolizumab in an ingestible device. In some embodiments, the A4 inhibitor is Tysabri.
(1) A4 inhibitor; (2) Vedolizumab subcutaneously. In some embodiments, the A4 inhibitor is Tysabri.
(1) anti-sense VCAM inhibitor; (2) Tysabri.
(1) anti-sense VCAM inhibitor; (2) Tysabri in an ingestible device.
(1) anti-sense VCAM inhibitor; (2) Vedolizumab.
(1) anti-sense VCAM inhibitor; (2) Vedolizumab in an ingestible device.
(1) anti-sense VCAM inhibitor; (2) Vedolizumab intravenously or subcutaneously.
(1) Cyclosporine; (2) vedolizumab.
(1) Cyclosporine; (2) vedolizumab in an ingestible device.
(1) Cyclosporine; (2) vedolizumab intravenously or subcutaneously.
(1) TNF inhibitor; (2) MADCAM inhibitor.
(1) TNF inhibitor; (2) MADCAM inhibitor in an ingestible device.
(1) TNF inhibitor; (2) B7 inhibitor.
(1) B7 inhibitor; TNF inhibitor.
(1) TNF inhibitor; (2) B7 inhibitor in an ingestible device.
(1) B7 inhibitor; TNF inhibitor in an ingestible device.
(1) TNF inhibitor; (2) B7 inhibitor intravenously or subcutaneously.
(1) B7 inhibitor; TNF inhibitor intravenously or subcutaneously.
(1) JAK inhibitor; (2) TNF inhibitor.
(1) JAK inhibitor; (2) TNF inhibitor in an ingestible device.
(1) JAK inhibitor; (2) TNF inhibitor intravenously or subcutaneously.
(1) TNF inhibitor; (2) JAK inhibitor
(1) TNF inhibitor; (2) JAK inhibitor in an ingestible device.
(1) TNF inhibitor; (2) JAK inhibitor orally.
(1) Neoregulin-4; (2) TNF inhibitor.
(1) Neoregulin-4; (2) TNF inhibitor in an ingestible device.
(1) Neoregulin-4; (2) TNF inhibitor intravenously or subcutaneously.
(1) Neoregulin-4; (2) vedolizumab.
(1) Neoregulin-4; (2) vedolizumab in an ingestible device.
(1) Neoregulin-4; (2) vedolizumab intravenously or subcutaneously.

(1) Neoregulin-4; (2) Stelara.

[1741] (1) Neoregulin-4; (2) Stelara in an ingestible device.
(1) Neoregulin-4; (2) Stelara intravenously or subcutaneously.
(1) Neoregulin-4; (2) JAK inhibitor.
(1) Neoregulin-4; (2) JAK inhibitor in an ingestible device.
(1) Neoregulin-4; (2) JAK inhibitor intravenously or subcutaneously.
(1) TNF inhibitor; (2) SiP inhibitor. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.
(1) TNF inhibitor; (2) S1P inhibitor orally. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.
(1) Stelara; (2) S1P inhibitor. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.
(1) Stelara; (2) S1P inhibitor orally. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.
(1) Vedolizumab; (2) S1P inhibitor. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.
(1) Vedolizumab; (2) S1P inhibitor orally. In some embodiments, the S1P inhibitor is ozanimod or etrasimod.

[1742] In some embodiments, the methods disclosed herein comprise administering (i) the immune modulator as disclosed herein, and (ii) a second agent orally, intravenously or subcutaneously, wherein the second agent in (ii) is the same immune modulator in (i); a different immune modulator; or an agent having a different biological target from the immune modulator.

[1743] In some embodiments, the methods disclosed herein comprise administering (i) the immune modulator in the manner disclosed herein, and (ii) a second agent orally, intravenously or subcutaneously, wherein the second agent in (ii) is an agent suitable for treating an inflammatory bowel disease.

[1744] In some embodiments, the immune modulator is administered prior to the second agent. In some embodiments, the immune modulator is administered after the second agent. In some embodiments, the immune modulator and the second agent are administered substantially at the same time. In some embodiments, the immune modulator is delivered prior to the second agent. In some embodiments, the immune modulator is delivered after the second agent. In some embodiments, the immune modulator and the second agent are delivered substantially at the same time.

[1745] In some embodiments, the second agent is an agent suitable for the treatment of an inflammatory disease or condition that arises in a tissue originating from the endoderm. In some embodiments, the second agent is administered intravenously. In some embodiments, the second agent is administered subcutaneously.

[1746] In some embodiments, delivery of the immune modulator to the location, such as delivery to the location by mucosal contact, results in systemic immunogenicity levels at or below systemic immunogenicity levels resulting from administration of the immune modulator systemically. In some embodiments comprising administering the immune modulator in the manner disclosed herein and a second agent systemically, delivery of the immune modulator to the location, such as delivery to the location by mucosal contact, results in systemic immunogenicity levels at or below systemic immunogenicity levels resulting from administration of the immune modulator systemically and the second agent systemically. In some embodiments, the method comprises administering the immune modulator in the manner disclosed herein and a second agent, wherein the amount of the second agent is less than the amount of the second agent when the immune modulator and the second agent are both administered systemically.

EXAMPLES

Example 1Preclinical Murine Colitis Model

Experimental Induction of Colitis

[1747] Colitis is experimentally induced to mice via the dextran sulfate sodium (DSS)-induced colitis model. This model is widely used because of its simplicity and many similarities with human ulcerative colitis. Briefly, mice are subjected to DSS via cecal catheterization, which is thought to be directly toxic to colonic epithelial cells of the basal crypts, for several days until colitis is induced.

Groups

[1748] Mice are allocated to one of seven cohorts, depending on the agent that is administered: [1749] 1. Control (no agent) [1750] 2. Adalimumab (2.5 mg/kg) [1751] 3. Adalimumab (5 mg/kg) [1752] 4. Adalimumab (10 mg/kg)

[1753] The control or agent is applied to a damaged mucosal surface of the bowel via administration through a cecal catheter at the dose levels described above.

[1754] Additionally, for each cohort, the animals are separated into two groups. One group receives a single dose of the control or agent on day 10 or 12. The other group receives daily (or similar) dosing of the control or agent.

Analysis

[1755] For each animal, efficacy is determined (e.g., by endoscopy, histology, etc.), and cytotoxic T-cell levels are determined in blood, feces, and tissue (tissue levels are determined after animal sacrifice). For tissue samples, levels HER2 are additionally determined, and the level of cytotoxic T cells is normalized to the level of HER2. Additionally, other cytokine levels are determined in tissue (e.g., phospho STAT 1, STAT 3 and STAT 5), in plasma (e.g., VEGF, VCAM, ICAM, IL-6), or both.

[1756] Pharmacokinetics are determined both systemically (e.g., in the plasma) and locally (e.g., in colon tissue). For systemic pharmacokinetic analysis, blood and/or feces is collected from the animals at one or more timepoints after administration (e.g., plasma samples are collected at 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and/or 8 hours after administration). Local/colon tissue samples are collected once after animal sacrifice.

Example 2aDevelopment of Preclinical Porcine Colitis Model

Experimental Induction of Colitis

[1757] Female swine weighing approximately 35 to 45 kg at study start are fasted at least 24 hours prior to intra-rectal administration of trinitrobenzene sulfonic acid (TNBS). Animals are lightly anesthetized during the dosing and endoscopy procedure. An enema to clean the colon is used, if necessary. One animal is administered 40 ml of 100% EtOH mixed with 5 grams of TNBS diluted in 10 ml of water via an enema using a ball-tipped catheter. The enema is deposited in the proximal portion of the descending colon just past the bend of the transverse colon. The TNBS is retained at the dose site for 12 minutes by use of two Foley catheters with 60-ml balloons placed in the mid-section of the descending colon below the dose site. A second animal is similarly treated, but with a solution containing 10 grams of TNBS. An Endoscope is employed to positively identify the dose site in both animals prior to TNBS administration. Dosing and endoscopy are performed by a veterinary surgeon

[1758] Seven (7) days after TNBS administration, after light anesthesia, the dose site and mucosal tissues above and below the dose site are evaluated by the veterinary surgeon using an endoscope. Pinch Biopsies are obtained necessary, as determined by the surgeon. Based on the endoscopy findings, the animals may be euthanized for tissue collection on that day, or may proceed on study pending the results of subsequent endoscopy exams for 1 to 4 more days. Macroscopic and microscopic alterations of colonic architecture, possible necrosis, thickening of the colon, and substantial histologic changes are observed at the proper TNBS dose.

[1759] Clinical signs (e.g., ill health, behavioral changes, etc.) are recorded at least daily during acclimation and throughout the study. Additional pen-side observations are conducted twice daily (once-daily on weekends). Body weight is measured for both animals Days 1 and 7 (and on the day of euthanasia if after Day 7).

[1760] On the day of necropsy, the animals are euthanized via injection of a veterinarian-approved euthanasia solution. Immediately after euthanasia in order to avoid autolytic changes, colon tissues are collected, opened, rinsed with saline, and a detailed macroscopic examination of the colon is performed to identify macroscopic finings related to TNBS-damage. Photos are taken. Tissue samples are taken from the proximal, mid, and distal transverse colon; the dose site; the distal colon; the rectum; and the anal canal. Samples are placed into NBF and evaluated by a board certified veterinary pathologist.

Example 2bPharmacokinetic/Pharmacodynamic and Bioavailability of Adalimumab After Topical Application

Groups

[1761] Sixteen (16) swine (approximately 35 to 45 kg at study start) are allocated to one of five groups:

[1762] 1. Vehicle Control: (3.2 mL saline); intra-rectal; (n=2)

[1763] 2. Treated Control: Adalimumab (40 mg in 3.2 mL saline); subcutaneous; (n=2)

[1764] 3. Adalimumab (low): Adalimumab (40 mg in 3.2 mL saline); intra-rectal; (n=4)

[1765] 4. Adalimumab (med): Adalimumab (80 mg in 3.2 mL saline); intra-rectal; (n=4)

[1766] 5. Adalimumab (high): Adalimumab (160 mg in 3.2 mL saline); intra-rectal; (n=4)

[1767] On Day 0, the test article is applied to a damaged mucosal surface of the bowel via intra-rectal administration or subcutaneous injection by a veterinary surgeon at the dose levels and volume described above.

Clinical Observations and Body Weight

[1768] Clinical observations are conducted at least once daily. Clinical signs (e.g., ill health, behavioral changes, etc.) are recorded on all appropriate animals at least daily prior to the initiation of experiment and throughout the study until termination. Additional clinical observations may be performed if deemed necessary. Animals whose health condition warrants further evaluation are examined by a Clinical Veterinarian. Body weight is measured for all animals Days 6, 0, and after the last blood collections.

Samples

[1769] Blood:

[1770] Blood is collected (cephalic, jugular, and/or catheter) into EDTA tubes during acclimation on Day-7, just prior to dose on Day 0, and 0.5, 1, 2, 4, 6, 8, 12, 24, and 48 hours post-dose. The EDTA samples are split into two aliquots and one is centrifuged for pharmacokinetic plasma and either analyzed immediately, or stored frozen (80 C.) for later pharmacokinetic analyses. The remaining sample of whole blood is used for pharmacodynamic analyses.

[1771] Feces:

[1772] Feces is collected Day 7, 0 and 0.5, 1, 2, 4, 6, 8, 12, 24 and 48 hours post-dose, and either analyzed immediately, or flash-frozen on liquid nitrogen and stored frozen at 70 C. pending later analysis of drug levels and inflammatory cytokines.

[1773] Tissue:

[1774] Immediately after euthanasia in order to avoid autolytic changes, colon tissues are collected, opened, rinsed with saline, and a detailed macroscopic examination of the colon is performed to identify macroscopic finings related to TNBS-damage. Triplicate samples of normal and damaged tissues are either analyzed immediately, or are flash-frozen on liquid nitrogen and stored frozen at 70 C. pending later analysis of drug concentration, inflammatory cytokines and histology.

[1775] Samples are analyzed for adalimumab levels (local mucosal tissue levels and systemic circulation levels), and for levels of inflammatory cytokines including TNF-alpha.

Terminal Procedures

[1776] Animals are euthanized as per the schedule in Table AA, where one animal each of Vehicle and Treated Control groups is euthanized at 6 and 48 hours post-dose, and one animal of each the adalimumab groups are euthanized at 6, 12, 24 and 48 hours post-dose. Animals are discarded after the last blood collection unless retained for a subsequent study.

TABLE-US-00029 TABLE AA Sample Days Hours General size Dose Route 7 6 5 4 3 2 1 0 0.5 1 2 4 6 8 12 24 48 Fast Food/Water ad oral libidum Observations clinical observations body weight Treatments (groups) TNBS intra (all animals) rectal 1. Vehicle control n = 2 1.6 mL intra saline rectal (vehicle) euthanized n = 1 n = 1 2. Treated control n = 2 40 mg in sub- 1.6 mL cutaneous saline euthanized n = 1 n = 1 3. Adalimumab (low) n = 4 40 mg in intra 1.6 mL rectal saline euthanized n = 1 n = 1 n = 1 n = 1 4. Adalimumab (med) n = 4 80 mg in intra 1.6 mL rectal saline euthanized n = 1 n = 1 n = 1 n = 1 5. Adalimumab (high) n = 4 160 mg in intra 1.6 mL rectal saline euthanized n = 1 n = 1 n = 1 n = 1 Adalimumab (required) 1200 Samples Blood cephalic, jugular or catheter Fecal rectal Tissue necropsy

Example 2cPharmacokinetic/Pharmacodynamic and Bioavailability of Adalimumab after Topical Application

Groups

[1777] DSS-induced colitis Yorkshire-Cross Farm Swine (approximately 5-10 kg at study start) are allocated to one of five groups: [1778] 1. Vehicle Control: (saline); intra-rectal; [1779] 2. Treated Control: Adalimumab (13 mg in saline); subcutaneous; [1780] 3. Adalimumab: Adalimumab (13 mg in saline); intra-rectal;

[1781] At t=0, the test article is applied to a damaged mucosal surface of the bowel via intra-rectal administration or subcutaneous injection by a veterinary surgeon at the dose levels and volume described above.

Clinical Observations

[1782] Clinical signs (e.g., ill health, behavioral changes, etc.) are recorded on all appropriate animals at least daily prior to the initiation of experiment and throughout the study until termination. Additional clinical observations may be performed if deemed necessary. Animals whose health condition warrants further evaluation are examined by a Clinical Veterinarian.

Samples

[1783] Blood:

[1784] Blood is collected (cephalic, jugular, and/or catheter) into EDTA tubes during acclimation on Day-7, just prior to dose on Day 0, and 12 hours post-dose. The EDTA samples are split into two aliquots and one is centrifuged for pharmacokinetic plasma and either analyzed immediately, or stored frozen (80 C.) for later pharmacokinetic analyses. The remaining sample of whole blood is used for pharmacodynamic analyses.

[1785] Feces:

[1786] Feces is collected Day 7, 0 and 12 hours post-dose, and either analyzed immediately, or flash-frozen on liquid nitrogen and stored frozen at 70 C. pending later analysis of drug levels and inflammatory cytokines.

[1787] Tissue:

[1788] Immediately after euthanasia (12 hours after dosing) in order to avoid autolytic changes, colon tissues are collected, opened, rinsed with saline, and a detailed macroscopic examination of the colon is performed to identify macroscopic finings related to DSS-damage. Triplicate samples of normal and damaged tissues are either analyzed immediately, or are flash-frozen on liquid nitrogen and stored frozen at 70 C. pending later analysis of drug concentration, inflammatory cytokines and histology.

[1789] Samples are analyzed for adalimumab levels (local mucosal tissue levels and systemic circulation levels), and for levels of inflammatory cytokines including TNF-alpha.

Terminal Procedures

[1790] Animals are euthanized at 12 hours post-dose.

Example 3. Comparison of Systemic versus Intracecal Delivery of an Anti-IL-12 Antibody

[1791] The objective of this study was to compare the efficacy of an IL-12 inhibitor (anti-IL-12 p40; anti-p40 mAb; BioXCell (Cat #: BE0051)), when dosed systemically versus intracecally, to the treat dextran sulfate sodium salt (DSS)-induced colitis in male C57Bl/6 mice.

Materials and Methods

Mice

[1792] Normal male C57Bl/6 mice between the ages of 6-8 weeks old, weighing 20-24 g, were obtained from Charles River Laboratories. The mice were randomized into thirteen groups of twelve animals and two groups of eight animals, and housed in groups of 6-8 per cage, and acclimatized for at least three days prior to entering the study. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour, with an automatic timer for a light/dark cycle of 12 hours on/off, and fed with Labdiet 5053 sterile rodent chow, with water administered ad libitum.

Cecal Cannulation

[1793] Animals were placed under isoflurane anesthesia, with the cecum exposed via a midline incision in the abdomen. A small point incision was made in the distal cecum where 1-2 cm of the cannula was inserted. The incision was closed with a purse string suture using 5-0 silk. An incision was then made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was then washed copiously with warmed saline prior to closing the abdominal wall. A small incision was also made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All animals received 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until recovery before returning to their cage. All animals received 0.6 mg/kg BID buprenorphine for the first 3 days, and Baytril at 10 mg/Kg every day for the first 5 days post surgery.

Induction of Colitis

[1794] Colitis was induced in male C57Bl/6 mice by exposure to 3% DSS drinking water (MP Biomedicals #0260110) from Day 0 to Day 5. Fresh DSS/water solutions were made again on Day 3 and any of the remaining original DSS solution will be discarded.

Assessment of Colitis

[1795] All animals were weighed daily and visually assessed for the presence of diarrhea and/or bloody stool at the time of dosing. The mice underwent two video endoscopies, one on day 10 and one on day 14, to assess colitis severity. Images were captured from each animal at the most severe region of disease identified during the endoscopy, and assessed using the rubric demonstrated in Table 1.1. Additionally, stool consistency was scored during the endoscopy using this rubric (Table 1.2) (0=Normal, well-formed pellet, 1=Loose stool, soft, staying in shape, 2=Loose stool, abnormal form with excess moisture, 3=Watery or diarrhea, 4=Bloody diarrhea). At necropsy, intestinal contents, peripheral blood, and tissue, and cecum/colon contents were collected for analysis.

TABLE-US-00030 TABLE 1.1 Endoscopy Scoring Score Description of Endoscopy Score 0 Normal 1 Loss of vascularity 2 Loss of vascularity and friability 3 Friability and erosions 4 Ulcerations and bleeding

TABLE-US-00031 TABLE 1.2 Stool Consistency Score Score Description of Stool Consistency 0 Normal, well-formed pellet 1 Loose stool, soft, staying in shape 2 Loose stool, abnormal form with excess moisture 3 Watery or diarrhea 4 Bloody diarrhea

Treatment of Colitis

[1796] Mice were treated with anti-IL-12 p40 during the acute phase of colitis due to its efficacy in the treatment of DSS-induced colitis. The test article was dosed at a volume of 0.1 mL/20 g from days 0 to 14. Anti-IL-12 p40 was administered intraperitoneally at a dose of 10 mg/kg every days, and intracecally at a dose of 10 mg/kg, either every 3 days or every day. There was also a lower dose of 1 mg/kg given every day intracecally. The control groups were not administered drugs, and the vehicles (sterile PBS) were administered the placebo drug intraperitoneally and intracecally every day. These drugs were given from days 5-14, which is 9 days of administration. A more detailed explanation of dosing and groups can be seen in Table 1.3.

TABLE-US-00032 TABLE 1.3 Groups of Animals # of Cecal Dose Dosing Group # Animals DSS Cannula Treatment (mg/kg) Route Schedule 1 8 males NO 2 8 males YES 3 12 males 3% DSS NO Vehicle PO QD (day 0-5) day 0-14 4 12 males 3% DSS YES Vehicle IC QD (day 0-5) day 0-14 5 12 males 3% DSS NO Anti-p40 10 IP Q3 (day 0-5) 0, 3, 6, 9, 12 6 12 males 3% DSS YES Anti-p40 10 IC Q3 (day 0-5) 0, 3, 6, 9, 12 7 12 males 3% DSS YES Anti-p40 10 IC QD (day 0-5) day 0-14 8 12 males 3% DSS YES Anti-p40 1 IC QD (day 0-5) day 0-14

Sample Collection

[1797] Intestinal contents, peripheral blood, and tissue were collected at sacrifice on day 14, as follows: at the end of each study period, mice were euthanized by CO.sub.2 inhalation immediately following endoscopy on day 14. The blood was collected via cardiac puncture into K.sub.2EDTA-coated tubes and centrifuged at 4000g for 10 minutes. The blood cell pellet was retained and snapped frozen. The resulting plasma was then split into two separate cryotubes, with 100 L in one tube and the remainder in the second. Plasma and cell pellet were also collected, flash frozen, and stored at 80 degrees Celsius.

[1798] The cecum and colon were removed from each animal and contents were collected, weighed, and snap frozen in separate cryovials. The colon was excised, rinsed, measured, weighed, and then trimmed to 6 cm in length and divided into 5 pieces. The most proximal 1 cm of colon was snapped frozen for subsequent bioanalysis of test article levels. Of the remaining 5 cm of colon, the most distal and proximal 1.5-cm sections was placed in formalin for 24 hours then transferred to 70% ethanol for subsequent histological evaluation. The middle 2-cm portion was bisected longitudinally and placed into two separate cryotubes, weighed, and snap frozen in liquid nitrogen.

Results

[1799] The data in FIG. 30 show that the DSS mice that were intracecally administered an anti-IL-12 p40 (IgG2A) antibody had decreased weight loss as compared to DSS mice that were intraperitoneally administered the anti-IL-12 p40 antibody.

[1800] The data in FIG. 31 show that the plasma concentration of the anti-IL-12 p40 antibody was decreased in DSS mice that were intracecally administered the anti-IL-12 p40 antibody as compared to DSS mice that were intraperitoneally administered the anti-IL-12 p40 antibody. The data in FIG. 32 show that the cecum and colon concentration of the anti-IL-12 p40 antibody is increased in DSS mice that were intracecally administered the anti-IL-12 p40 antibody as compared to the DSS mice that were intraperitoneally administered the anti-IL-12 p40 antibody.

[1801] The data in FIGS. 33 and 34 show that the anti-IL-12 p40 antibody is able to penetrate colon tissues (the lumen superficial, lamina propria, submucosa, and tunica muscularis/serosa) in DSS mice intracecally administered the anti-IL-12 p40 antibody, while the anti-IL-12 p40 antibody did not detectably penetrate the colon tissues of DSS mice intraperitoneally administered the anti-IL-12 p40 antibody. The data in FIG. 35 also show that the ratio of the concentration of anti-IL-12 p40 antibody in colon tissue to the concentration of the anti-IL-12 p40 antibody in plasma is increased in DSS mice intracecally administered the anti-IL-12 p40 antibody as compared to the ratio in DSS mice intraperitoneally administered the anti-IL-12 p40 antibody.

[1802] The data in FIG. 36 show that the concentration of IL-1 in colon tissue is decreased in DSS mice intracecally administered the anti-IL-12 p40 antibody as compared to the concentration of IL-1 in colon tissue in DSS mice intraperitoneally administered the anti-IL-12 p40 antibody. The data in FIG. 37 show that the concentration of IL-6 in colon tissue is decreased in DSS mice intracecally administered the anti-IL-12 p40 antibody as compared to the concentration of IL-6 in colon tissue in DSS mice intraperitoneally administered the anti-IL-12 p40 antibody. The data in FIG. 38 show that the concentration of IL-17A in colon tissue is decreased in DSS mice intracecally administered the anti-IL-12 p40 antibody as compared to the concentration of IL-17A in colon tissue in DSS mice intraperitoneally administered the anti-IL-12 p40 antibody.

[1803] No significant differences in clinical observations or gastrointestinal-specific adverse effects, including stool consistency and/or bloody stool, were observed due to cannulation or intra-cecal treatments when compared with vehicle. No toxicity resulting from the treatments was reported. A significant reduction in body weight-loss (AUC) was found in groups treated with anti-IL-12 p40 antibody (10 mg/kg and 1 mg/kg, QD) via intra-cecal delivery when compared with vehicle control and intraperitoneal delivery (10 mg/kg, Q3D). The immunohistochemistry staining in anti-IL-12 p40 antibody (10 mg/kg, QD) treatment groups showed penetration of the antibody in all layers of colon tissue, including lumen mucosa, lamina propria, submucosa, tunica muscularis, via intra-cecal delivery. The distribution of anti-IL-12 p40 antibody was found in all segments of the colon, however, higher levels were detected in the proximal region. A significantly higher mean concentration of anti-IL-12 p40 antibody was found in the gastrointestinal contents and colon tissues when delivered via intra-cecal administration (Anti-p40: 10 mg/kg and 1 mg/kg, QD) compared with intraperitoneal administration (anti-p40: 10 mg/kg, Q3D). The blood level of anti-IL-12 p40 antibody was significantly higher when delivered via intraperitoneal administration (Q3D) as compared to intra-cecal administration (Q3D & QD). The concentrations of inflammatory cytokines, including IL-1, IL-6, and IL-17, were significantly reduced by anti-IL-12 p40 antibody (10 mg/kg, QD) treatment when delivered via intra-cecal administration as compared to vehicle controls.

[1804] In sum, these data show that the compositions and devices provided herein can suppress the local immune response in the intestine, while having less of a suppressive effect on the systemic immune response of an animal. These data also suggest that the presently claimed compositions and devices will provide for treatment of colitis and other pro-inflammatory disorders of the intestine.

Example 4. Comparison of Systemic Versus Intracecal Delivery of an Anti-Integrin 47 Antibody

[1805] The objective of this study was to compare the efficacy of an integrin inhibitor (anti-integrin 47; anti-LPAM1; DATK-32 mAb; BioXCell (Cat #: BE0034)) when dosed systemically versus intracecally for treating dextran sulfate sodium salt (DSS)-induced colitis in male C57Bl/6 mice.

Materials and Methods

Mice

[1806] Normal male C57Bl/6 mice between the ages of 6-8 weeks old, weighing 20-24 g, were obtained from Charles River Laboratories. The mice were randomized into thirteen groups of twelve animals and two groups of eight animals, and housed in groups of 6-8 per cage, and acclimatized for at least three days prior to entering the study. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour, with an automatic timer for a light/dark cycle of 12 hours on/off, and fed with Labdiet 5053 sterile rodent chow, with water administered ad libitum.

Cecal Cannulation

[1807] The animals were placed under isoflurane anesthesia, with the cecum exposed via a midline incision in the abdomen. A small point incision was made in the distal cecum where 1-2 cm of the cannula was inserted. The incision was closed with a purse string suture using 5-0 silk. An incision was then made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was then washed copiously with warmed saline prior to closing the abdominal wall. A small incision was also made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All animals received 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until recovery before returning to their cage. All animals received 0.6 mg/kg BID buprenorphine for the first 3 days, and Baytril at 10 mg/Kg every day for the first 5 days post-surgery.

Induction of Colitis

[1808] Colitis was induced in male C57Bl/6 mice by exposure to 3% DSS drinking water (MP Biomedicals #0260110) from day 0 to day 5. Fresh DSS/water solutions were made again on day 3 and any of the remaining original DSS solution will be discarded.

Assessment of Colitis

[1809] All animals were weighed daily and visually assessed for the presence of diarrhea and/or bloody stool at the time of dosing. Mice underwent two video endoscopies, one on day 10 and one on day 14, to assess colitis severity. Images were captured from each animal at the most severe region of disease identified during the endoscopy, and assessed using the rubric demonstrated in Table 2.1. Additionally, stool consistency was scored during the endoscopy using this rubric (Table 2.2) (0=Normal, well-formed pellet, 1=Loose stool, soft, staying in shape, 2=Loose stool, abnormal form with excess moisture, 3=Watery or diarrhea, 4=Bloody diarrhea). At necropsy, intestinal contents, peripheral blood and tissue, and cecum/colon contents were collected for analysis.

TABLE-US-00033 TABLE 2.1 Endoscopy Score Score Description of Endoscopy Score 0 Normal 1 Loss of vascularity 2 Loss of vascularity and friability 3 Friability and erosions 4 Ulcerations and bleeding

TABLE-US-00034 TABLE 2.2 Stool Consistency Score Score Description of Stool Consistency 0 Normal, well-formed pellet 1 Loose stool, soft, staying in shape 2 Loose stool, abnormal form with excess moisture 3 Watery or diarrhea 4 Bloody diarrhea

Treatment of Colitis

[1810] Mice were treated with DATK32 during the acute phase of colitis due to its efficacy in the treatment of DSS-induced colitis. The test article was dosed at a volume of 0.1 mL/20 g from days 0 to 14. DATK32 was administered intraperitoneally at a dose of 25 mg/kg every 3 days, and intracecally at a dose of 25 mg/kg, either every 3 days or every day. There was also a lower dose of 5 mg/kg given every day intracecally. The control groups were not administered drugs, and the vehicle (sterile PBS) was administered as the placebo drug intraperitoneally and intracecally every day. These drugs were given from days 5-14, which is 9 days of administration. A more detailed explanation of dosing and groups can be seen in Table 2.3.

TABLE-US-00035 TABLE 2.3 Groups of Mice # of Cecal Dose Dosing Group # Animals DSS Cannula Treatment (mg/kg) Route Schedule 1 8 males NO 2 8 males YES 3 12 males 3% DSS NO Vehicle PO QD (day 0-5) day 0-14 4 12 males 3% DSS YES Vehicle IC QD (day 0-5) day 0-14 9 12 males 3% DSS NO DATK32 25 IP Q3 (day 0-5) 0, 3, 6, 9, 12 10 12 males 3% DSS YES DATK32 25 IC Q3 (day 0-5) 0, 3, 6, 9, 12 11 12 males 3% DSS YES DATK32 25 IC QD (day 0-5) day 0-14 12 12 males 3% DSS YES DATK32 5 IC QD (day 0-5) day 0-14

Sample Collection

[1811] Intestinal contents, peripheral blood, and tissue were collected at sacrifice on day 14, as follows: at the end of each study period, mice were euthanized by CO.sub.2 inhalation immediately following endoscopy on day 14. The blood was collected via cardiac puncture into K.sub.2EDTA-coated tubes and centrifuged at 4000g for 10 minutes. The blood cell pellet was retained and snapped frozen. The resulting plasma was then split into two separate cryotubes, with 100 L in one tube and the remainder in the second. Plasma and the cell pellet were also collected, flash frozen, and stored at 80 degrees Celsius. An ELISA was used to determine the level of rat IgG2A.

[1812] The cecum and colon were removed from each animal and contents were collected, weighed, and snap frozen in separate cryovials. The colon was excised, rinsed, measured, weighed, and then trimmed to 6 cm in length and divided into 5 pieces. The most proximal 1 cm of colon was snapped frozen for subsequent bioanalysis of anti-DATK32 levels. Of the remaining 5 cm of colon, the most distal and proximal 1.5-cm sections was placed in formalin for 24 hours then transferred to 70% ethanol for subsequent histological evaluation. The middle 2-cm portion was bisected longitudinally and placed into two separate cryotubes, weighed, and snap frozen in liquid nitrogen.

[1813] There was an additional collection of 100 L of whole blood from all animals and processed for FACS analysis of 4 and 7 expression on T-helper memory cells. Tissue and blood were immediately placed in FACS buffer (lx PBS containing 2.5% fetal calf serum) and analyzed using the following antibody panel (Table 2.4).

TABLE-US-00036 TABLE 2.4 Fluorophore Labelled Antibodies Used in FACS Analysis Antibody Target Flurochrome Purpose CD4 APC-Vio770 Defines T-Helper Cells CD44 VioBlue Memory/Naive Discrimination CD45RB FITC Memory/Naive Discrimination 4 APC Defines T-helper memory subset of interest 7 PE Defines T-helper memory subset of interest CD16/32 Fc Block

Results

[1814] The data in FIG. 39 show decreased weight loss in DSS mice intracecally administered DATK antibody as compared to DSS mice that were intraperitoneally administered the DATK antibody. The data in FIG. 40 show that DSS mice intracecally administered DATK antibody have a decreased plasma concentration of DATK antibody as compared to DSS mice that were intraperitoneally administered DATK antibody. The data in FIGS. 41 and 42 show that DSS mice intracecally administered DATK antibody have an increased concentration of DATK antibody in the cecum and colon content as compared to DSS mice intraperitoneally administered DATK antibody. The data in FIGS. 43 and 44 show that DSS mice intracecally administered DATK antibody have an increased concentration of DATK antibody in colon tissue as compared to DSS mice intraperitoneally administered DATK antibody. The data in FIGS. 45 and 46 show an increased level of penetration of DATK antibody into colon tissue in DSS mice intracecally administered the DATK antibody as compared to an intracecal vehicle control (PBS). The data in FIG. 47 show that DSS mice intracecally administered DATK antibody have an increased ratio of the concentration of DATK antibody in colon tissue to the plasma concentration of the DATK antibody, as compared to the same ratio in DSS mice intraperitoneally administered the DATK antibody.

[1815] The data in FIG. 48 show that DSS mice intracecally administered the DATK antibody have an increased percentage of blood Th memory cells as compared to DSS mice intraperitoneally administered the DATK antibody.

[1816] No significant differences in clinical observations or gastrointestinal-specific adverse effects, including stool consistency and/or bloody stool, were observed due to cannulation or intra-cecal treatments when compared with vehicle. No toxicity resulting from the treatments was reported. A significant reduction in body weight-loss was also found with DATK32 (5 mg/kg, QD) treatment (IC) when compared to vehicle control at the endpoint (day 14). The immunohistochemistry staining in DATK32 (25 mg/kg, QD) treatment groups showed penetration of DATK32 in all layers of colon tissue, including lumen mucosa, lamina propria, submucosa, tunica muscularis, via intra-cecal delivery. The distribution of DATK32 was found in all segments of the colon, however, higher levels were detected in the proximal region. A significantly higher mean concentration of DATK32 was found in gastrointestinal contents and colon tissues when delivered via intra-cecal administration (DATK32: 25 mg/kg and 5 mg/kg, QD) as compared to intraperitoneal administration (DATK32: 25 mg/kg, Q3D). The blood level of DATK32 was significantly higher when delivered via intraperitoneal administration (Q3D) as compared to intra-cecal administration (Q3D & QD). The pharmacokinetics of DATK32 (25 mg/kg, QD) showed significantly higher mean concentrations of DATK32 when delivered via intra-cecal administration at 1, 2, and 4 h post-dose in the gastrointestinal contents, and 1, 2, 4 and 24 h in colon tissue as compared with the mean concentrations of DATK32 following intraperitoneal administration. The mean number of gut-homing T cells (Th memory cells) was significantly higher in the blood of groups treated with DATK32 via intra-cecal administration (QD 25 mg/kg and QD 5 mg/kg) as compared to the groups treated with DATK32 via intraperitoneal administration (Q3D 25 mg/kg). The mean number of Th memory cells was significantly lower in the Peyer's Patches of groups treated with DATK32 via intra-cecal administration (QD 25 mg/kg and 5 mg/kg) as compared to the groups treated with DATK32 via intraperitoneal administration (Q3D 25 mg/kg). The mean number of Th memory cells in mesenteric lymph nodes (MLN) was significantly lower in groups treated with DATK32 via intra-cecal administration (QD and Q3D 25 mg/kg and QD 5 mg/kg) as compared to the groups treated with DATK32 via intraperitoneal administration (Q3D 25 mg/kg).

[1817] In sum, these data show that the compositions and devices provided herein can suppress the local immune response in the intestine, while having less of a suppressive effect on the systemic immune response of an animal. These data also show that the release of DATK-32 antibody in the colon can result in a suppression of leukocyte recruitment and may provide for the treatment of colitis and other pro-inflammatory diseases of the intestine.

Example 5. An Assessment of DATK32 Bio-Distribution Following Intracecal Administration in Male C57Bl/6 Mice

[1818] The objective of this study is to assess DATK32 bio-distribution when dosed intracecally in male C57Bl/6 mice. A minimum of 10 days prior to the start of the experiment a cohort of animals will undergo surgical implantation of a cecal cannula. A sufficient number of animals will undergo implantation to allow for 24 cannulated animals to be enrolled in the main study (e.g., 31 animals). Animals were dosed with vehicle or test article via intracecal injection (IC) on Day 0 as indicated in Table 3. Animals from all groups were sacrificed for terminal sample collection three hours following test article administration.

Materials and Methods

Mice

[1819] Normal male C57Bl/6 mice between the ages of 6-8 weeks old, weighing 20-24 g, were obtained from Charles River Laboratories. The mice were randomized into two groups of twelve animals, and housed in groups of 12 per cage, and acclimatized for at least three days prior to entering the study. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour, with an automatic timer for a light/dark cycle of 12 hours on/off, and fed with Labdiet 5053 sterile rodent chow, with water administered ad libitum.

Cecal Cannulation

[1820] The animals were placed under isoflurane anesthesia, with the cecum exposed via a midline incision in the abdomen. A small point incision was made in the distal cecum where 1-2 cm of the cannula was inserted. The incision was closed with a purse string suture using 5-0 silk. An incision was then made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was then washed copiously with warmed saline prior to closing the abdominal wall. A small incision was also made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All animals received 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until recovery before returning to their cage. All animals received 0.6 mg/kg BID buprenorphine for the first 3 days, and Baytril at 10 mg/Kg every day for the first 5 days post-surgery.

[1821] Dosing

[1822] Animals were dosed IC at a volume of 0.075 mL/animal on Days 0 as indicated in Table 3.

Sacrifice

[1823] All animals were euthanized by CO.sub.2 inhalation three hours after dosing on Day 0.

Sample Collection

[1824] Terminal blood was collected and prepared for plasma using K.sub.2EDTA as the anti-coagulant. The plasma will be split into two cryotubes, with 50 L in one tube (PK analysis) and the remainder in another (other). Both samples were flash-frozen in liquid nitrogen. Plasma was stored at 80 C. for downstream analysis. Mesenteric lymph nodes (mLN) were collected, weighed, and flash-frozen in liquid nitrogen. Mesenteric lymph nodes were stored at 80 C. for downstream analysis. The small intestine was excised and rinsed, and the most distal 1 cm of ilium was dissected, weighed, and flash-frozen in liquid nitrogen. The samples were stored at 80 C. for downstream analysis. The cecum and colon were removed from each animal and contents collected, weighed, and snap frozen in separate cryovials. The samples were stored at 80 C. for downstream analysis. The colon was rinsed, and the most proximal 1 cm of colon was weighed and flash-frozen in liquid nitrogen. The snap frozen tissues were stored at 80 C.

TABLE-US-00037 TABLE 3 Study Design Terminal No Collections Group Animals Treatment Route Schedule Day 0 1 12 Vehicle IC Day 0 ** Blood (plasma) (PBS) Small 2 12 DATK32 intestine mLN (625 g)* Colon Colon Contents Cecum Contents *Per mouse. TA was administered in 0.075 mL/animal. DATK32 was delivered in sterile PBS. ** Animals were dosed on Day 0 and collections were performed 3 hours later.

Results

[1825] The data in FIGS. 63A-F show no significant differences in clinical observations. No gastrointestinal-specific or adverse effects were found in the group administered DATK32 via intra-cecal administration as compared to the group administered a vehicle control. No toxicity resulting from the treatments was reported. The level of DATK32 in the group intracecally administered DATK32 was significantly higher in cecum and colon content, and colon tissue compared to the group administered a vehicle control at 3h post-dose. A small amount of DATK32 was also detected in plasma, small intestine, and mesenteric lymph node in the group intra-cecally administered DATK32.

Example 6. Pharmacokinectics/Pharmacodynamics and Bioavailability of Adalimumab When Applied to a TNBS-damaged Mucosal Surface (Induced Colitis) in Swine

[1826] The purpose of this non-Good Laboratory Practice (GLP) study was to explore the PK/PD, and bioavailability of adalimumab when applied to a TNBS-damaged mucosal surface (induced colitis) in Yorkshire-Cross farm swine, and to determine an appropriate dose and frequency for studies where a drug will be delivered by the ingestible device system. The ingestible device system will be capable of delivering a TNF inhibitor (adalimumab) topically and locally to damaged mucosa in human patients with inflammatory bowel disease (IBD). The TNBS-induced colitis model was validated when a single administration on Day 1 of 40 mL of 100% ethanol (EtOH) mixed with 5 grams of TNBS diluted in 10 mL of water via an enema using a rubber catheter resulted in the intended reproducible induction of damaged mucosal surface (induced colitis) in Yorkshire-Cross farm swine.

[1827] This study investigated whether topical delivery of adalimumab would result in increased local mucosal tissue levels with limited drug reaching systemic circulation, as compared to subcutaneous administration; whether local mucosal tissue levels of drug would be greater in damaged tissues when compared to normal tissues; whether increasing the dose of drug would result in increased mucosal tissue levels in local and distal TNBS-damaged tissues; and whether topical delivery of adalimumab would result in reductions in inflammatory cytokines such as TNF- in damaged tissues, feces, and possibly blood.

[1828] All animals were subjected to intra-rectal administration of trinitrobenzene sulfonic acid (TNBS) to induce chronic colitis on day 2. All animals were fasted prior to colitis induction. Bedding was removed and replaced with rubber mats on day 3 to prevent ingestion of straw bedding material. The dose was 40 mL of 100% EtOH mixed with 5 grams of TNBS diluted in 10 mL of water, then instilled into the colon intra-rectally using a flexible gavage tube by a veterinary surgeon (deposited in a 10-cm portion of the distal colon and proximal rectum, and retained for 12 minutes by use of two Foley catheters with 60-mL balloons). Approximately 3 days after induction, macroscopic and microscopic alterations of colonic architecture were apparent: some necrosis, thickening of the colon, and substantial histologic changes were observed (FIGS. 49 and 50). The study employed 15 female swine (approximately 35 to 45 kg at study start) allocated to one of five groups. Group 1 employed three animals that were the treated controls. Each animal in Group 1 was administered adalimumab by subcutaneous injection at 40 mg in 0.8 mL saline. Groups 2, 3, 4, and 5 employed 3 animals in each group. Animals in these groups were administered intra-rectal adalimumab at 40 mg in 0.8 mL saline. The test drug (adalimumab) was administered to all groups on study day 1. The intra-rectal administrations (Groups 2-5) were applied to damaged mucosal surface of the bowel vial intra-rectal administration by a veterinary surgeon. Blood (EDTA) was collected from all animals (cephalic, jugular, or catheter) on day 3 (n=15), 1 (n=15), and 6 (n=15), 12 (n=12), 24 (n=9), and 48 (n=6) hours post-dose (87 bleeds total). The EDTA samples were split into two aliquots, and one was centrifuged for PK plasma, and stored frozen (80 C.) for PK analyses and reporting. Fecal samples were collected for the same time-points (87 fecal collections). Fecal samples were flash-frozen in liquid nitrogen and then stored at 80 C. for analysis of drug levels and inflammatory cytokines. Groups 2, 3, 4, and 5 were euthanized and subjected to gross necropsy and tissue collection 6, 12, 24, and 48 hours post-dose, respectively. Group 1 was similarly euthanized and necropsied 48 hours post-dose. The animals were euthanized via injection of a veterinarian-approved euthanasia solution as per the schedule. Immediately after euthanasia in order to avoid autolytic changes, colon tissues were collected, opened, rinsed with saline, and a detailed macroscopic examination of the colon were performed to identify macroscopic findings related to TNBS-damage. Tissue samples were taken from the proximal, mid, and distal transverse colon; the dose site; and the distal colon. Each tissue sample was divided into two approximate halves; one tissue section was placed into 10% neutral buffered formalin (NBF) and evaluated by a Board certified veterinary pathologist, and the remaining tissue section was flash frozen in liquid nitrogen and stored frozen at 80 C. Clinical signs (ill health, behavioral changes, etc.) were recorded daily beginning on day 3. Additional pen-side observations were conducted once or twice daily. Animals observed to be in ill health were examined by a veterinarian. Body weight was measured for all animals on day 3, and prior to scheduled euthanasia. Table 4.1, depicted below, shows the study design.

Materials and Methods

Test Article

[1829] Adalimumab (EXEMPTIA) is a Tumour Necrosis Factor (TNF) inhibitor. A single dose was pre-filled in a syringe (40 mg in a volume of 0.8 mL).

TABLE-US-00038 TABLE 4.1 Study Design Table Sample Days Hours General size Dose Route 3 2 1 1 0.5 1 2 4 6 8 12 24 48 Fast Food/Water ad libidum oral Observations clinical observations body weight Treatments (groups) TNBS intra (all animals) rectal 1. Treated control n = 3 40 mg in sub- 0.8 mL cutaneous saline euthanized n = 3 2. Adalimumab n = 3 40 mg in intra 0.8 mL rectal saline euthanized n = 3 3. Adalimumab n = 3 40 mg in intra 0.8 mL rectal saline euthanized n = 3 4. Adalimumab n = 3 40 mg in intra 0.8 mL rectal saline euthanized n = 3 5. Adalimumab n = 3 40 mg in intra 0.8 mL rectal saline euthanized n = 3 Adalimumab (required) 600 Samples PBMCs cephalic, jugular or catheter Serum cephalic, jugular or catheter Fecal rectal Tissue necropsy Analysis Histopathology 1 location 4 locations inflammed 45 180 H&E normal 45 180 H&E Blood adalimumab 57 pbl 15 15 12 9 6 TNF 87 pbl 15 15 15 15 12 9 6 Feces adalimumab 57 pbl 15 15 12 9 6 TNF 87 pbl 15 15 15 15 12 9 6 Tissue Inflammed adalimumab 45 180 pbl 3 3 3 6 TNF 45 180 pbl 3 3 3 6 HER2 45 180 pbl 3 3 3 6 Normal adalimumab 45 180 pbl 3 3 3 6 TNF 45 180 pbl 3 3 3 6 HER2 45 180 pbl 3 3 3 6

Results

[1830] While subcutaneously administered adalimumab was detected at all times points tested in plasma, topically administered adalimumab was barely detectable in plasma (FIGS. 51 and 52). Both topical delivery and subcutaneous delivery of adalimumab resulted in reduced levels of TNF- in colon tissue of TNBS-induced colitis animals, yet topical delivery of adalimumab was able to achieve a greater reduction in TNF- levels (FIGS. 53 and 54).

[1831] Either subcutaneous or intra-rectal administration of adalimumab was well tolerated and did not result in death, morbidity, adverse clinical observations, or body weight changes. A decreased level of total TNBS-related inflammatory response was observed by adalimumab treatment via intra-rectal administration when applied to the damaged mucosal surface of the bowel when compared to subcutaneous delivery. A significantly higher concentration of adalimumab was measured in blood following subcutaneous delivery as compared to the blood concentration following intra-rectal administration. Intra-rectal administration of adalimumab decreased the total and normalized TNF concentration over time (6-48h) and was more effective at reducing TNF at the endpoint (48h) as compared to groups administered adalimumab subcutaneously.

[1832] In sum, these data show that the compositions and devices provided herein can suppress the local immune response in the intestine, while having less of a suppressive effect on the systemic immune response of an animal. For example, these data show that intracecal administration of adalimumab using a device as described herein can provide for local delivery of adalimumab to the site of disease, without suppressing the systemic immune response. These data also show that local administration of adalimumab using a device as described herein can result in a significant reduction of the levels of TNF in diseases animals.

Example 7. Comparison of Systemic Versus Intracecal Delivery of Cyclosporine A

[1833] The objective of this study was to compare the efficacy of an immunosuppressant agent (cyclosporine A; CsA) when dosed systemically versus intracecally to treat dextran sulfate sodium salt (DSS)-induced colitis in male C57Bl/6 mice.

Experimental Design

[1834] A minimum of 10 days prior to the start of the experiment a cohort of animals underwent surgical implantation of a cecal cannula. A sufficient number of animals underwent implantation to allow for 44 cannulated animals to be enrolled in the main study (e.g., 76 animals). Colitis was induced in 60 male C5Bl/6 mice by exposure to 3% DSS-treated drinking water from day 0 to day 5. Two groups of eight additional animals (cannulated and non-cannulated) served as no-disease controls (Groups 1 and 2). Animals were dosed with cyclosporine A via intraperitoneal injection (IP), oral gavage (PO), or intracecal injection (IC) from day 0 to 14 as indicated in Table 5.1. All animals were weighed daily and assessed visually for the presence of diarrhea and/or bloody stool at the time of dosing. Mice underwent video endoscopy on days 10 and 14 to assess colitis severity. Images were captured from each animal at the most severe region of disease identified during endoscopy. Additionally, stool consistency was scored during endoscopy using the parameters defined in Table 5.2. Following endoscopy on day 14, animals from all groups were sacrificed and underwent terminal sample collection.

[1835] Specifically, animals in all treatment groups dosed on day 14 were sacrificed at a pre-dosing time point, or 1, 2, and 4 hours after dosing (n=3/group/time point). Terminal blood was collected via cardiac puncture and prepared for plasma using K.sub.2EDTA as the anti-coagulant. The blood cell pellet was retained and snap frozen while the resulting plasma was split into two separate cryotubes, with 100 L in one tube and the remainder in the second. Additionally, the cecum and colon were removed from all animals; the contents were collected, weighed, and snap frozen in separate cyrovials. The colon was then rinsed, measured, weighed, and then trimmed to 6 cm in length and divided into five pieces. The most proximal 1 cm of colon was snap frozen for subsequent bioanalysis of cyclosporine A levels. Of the remaining 5 cm of colon, the most distal and proximal 1.5-cm sections were each placed in formalin for 24 hours, then transferred to 70% ethanol for subsequent histological evaluation. The middle 2-cm portion was bisected longitudinally and placed into two separate cryotubes, weighed, and snap frozen in liquid nitrogen. All plasma and frozen colon tissue were stored at 80 C. for selected end point analysis. For all control animals in Groups 1-4, there was an additional collection of 100 L of whole blood from all animals which was then processed for FACS analysis of 4 and 7 expression on TH memory cells. The details of the study are shown in Table 5.1.

TABLE-US-00039 TABLE 5.1 Study Design Group 1 2 3 4 13 14 15 Number Number of 8 8 12 12 12 12 12 Animals Cecal NO YES NO YES NO YES YES Cannula DSS N/A N/A 3% DSS on Day 0 to Day 5 Treatment none none vehicle vehicle CsA CsA CsA Dose N/A N/A N/A N/A 10 10 3 (mg/kg) Route N/A N/A N/A N/A PO IC IC Dosing N/A N/A QD: Day QD: Day QD: Day QD: Day QD: Day Schedule 0 to 14 0 to 14 0 to 14 0 to 14 0 to 14 Endoscopy Schedule* Days 10 and 14 Endpoints Endoscopy, Colon weight/length, stool score Day 14 Terminal Collection (all groups): Cecal contents, colon contents, plasma, and colon tissue FACS analysis collection of Groups 1-4: Whole blood for the following FACS panel: CD4, CD44, CD45RB, 4, 7, CD16/32 PK N = 3/ time points Sacrifice At pre-dose and 1, 2, and 4 hours post-dosing (Day 14) *Animals were dosed once (QD) on Day 14 and plasma collected (K2EDTA) at pre-dosing, 1, 2, and 4 hours post-dosing from n = 3/group/time point. Each collection was terminal.

Experimental Procedures

Cecal Cannulation

[1836] Animals were placed under isoflurance anesthesia, and the cecum exposed via a mid-line incision in the abdomen. A small point incision was made in the distal cecum through which 1-2 cm of the cannula was inserted. The incision was closed with a purse-string suture using 5-0 silk. An incision was made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was washed copiously with warmed saline prior to closing the abdominal wall. A small incision was made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All animals received 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until fully recovered before returning to the cage. All animals received buprenorphine at 0.6 mg/kg BID for the first 3 days, and Baytril at 10 mg/kg QD for the first 5 days following surgery.

Disease Induction

[1837] Colitis was induced on day 0 via addition of 3% DSS (MP Biomedicals, Cat #0260110) to the drinking water. Fresh DSS/water solutions were made on day 3 and any of the remaining original DSS solution was discarded.

Dosing

[1838] Animals were dosed by oral gavage (PO), intraperitoneal injection (P), or intracecal injection (IC) at a volume of 0.1 mL/20 g on days 0 to 14 as indicated in Table 5.1.

Body Weight and Survival

[1839] Animals were observed daily (weight, morbidity, survival, presence of diarrhea, and/or bloody stool) in order to assess possible differences among treatment groups and/or possible toxicity resulting from the treatments.

Animals Found Dead or Moribund

[1840] Animals were monitored on a daily basis and those exhibiting weight loss greater than 30% were euthanized, and samples were not collected from these animals.

Endoscopy

[1841] Each mouse underwent video endoscopy on days 10 and 14 using a small animal endoscope (Karl Storz Endoskope, Germany) under isoflurane anesthesia. During each endoscopic procedure still images as well as video were recorded to evaluate the extent of colitis and the response to treatment. Additionally, we attempted to capture an image from each animal at the most severe region of disease identified during endoscopy. Colitis severity was scored using a 0-4 scale (0=normal; 1=loss of vascularity; =loss of vascularity and friability; 3=friability and erosions; 4=ulcerations and bleeding). Additionally, stool consistency was scored during endoscopy using the parameters defined in Table 5.2.

TABLE-US-00040 TABLE 5.2 Stool Consistency Score Description 0 Normal, well-formed pellet 1 Loose stool, soft, staying in shape 2 Loose stool, abnormal form with excess moisture 3 Watery or diarrhea 4 Bloody diarrhea

Tissue/Blood for FACS

[1842] Tissue and blood were immediately placed in FACS buffer (lx phosphate-buffered saline (PBS) containing 2.5% fetal calf serum (FCS)) and analyzed using the antibody panel in Table 5.3.

TABLE-US-00041 TABLE 5.3 FACS Antibody Panel Antibody Target Fluorochrome Purpose CD4 APC-Vio770 Defines T.sub.H cells CD44 VioBlue Memory/Nave discrimination CD45RB FITC Memory/Nave discrimination 4 APC Defines T.sub.H-memory subset of interest 7 PE Defines T.sub.H-memory subset of interest CD16/32 Fc block

Results

[1843] The data in FIG. 55 show a decrease in weight loss is observed in DSS mice intracecally administered cyclosporine A as compared to DSS mice orally administered cyclosporine A. The data in FIG. 56 show a decrease in plasma concentration of cyclosporine A in DSS mice intracecally administered cyclosporine A as compared to DSS mice orally administered cyclosporine A. The data in FIGS. 57-59 show an increased concentration of cyclosporine A in the colon tissue of DSS mice intracecally administered cyclosporine A as compared to the concentration of cyclosporine A in the colon tissue of DSS mice orally administered cyclosporine A.

[1844] The data in FIG. 60 show that DSS mice intracecally administered cyclosporine A have an increased concentration of IL-2 in colon tissue as compared to DSS mice orally administered cyclosporine A. The data in FIG. 61 show that DSS mice intracecally administered cyclosporine A have a decreased concentration of IL-6 in colon tissue as compared to DSS mice orally administered cyclosporine A.

[1845] In sum, these data show that the compositions and devices provided herein can suppress the local immune response in the intestine, while having less of a suppressive effect on the systemic immune response of an animal. For example, these data demonstrate that the present compositions and devices can be used to release cyclosporine A to the intestine and that this results in a selective immune suppression in the colon, while having less of an effect on the immune system outside of the intesting. These data also suggest that the present compositions and devices will provide for the treatment of colitis and other pro-inflammatory disorders of the intestine.

Example 8. Bellows Testing: Drug Stability Bench Test

[1846] Experiments were run to evaluate the effects that bellows material would have on the function of a drug used as the dispensable substance. The experiments also evaluated the effects on drug function due to shelf life in the bellows.

[1847] The adalimumab was loaded into simulated device jigs containing either tapered silicone bellows or smooth PVC bellows and allowed to incubate for 4, 24, or 336 hours at room temperature while protected from light. FIG. 64 illustrates the tapered silicone bellows, and FIG. 65 illustrates the tapered silicone bellows in the simulated device jig. FIG. 66 illustrates the smooth PVC bellows, and FIG. 67 illustrates the smooth PVC in the simulated device jig.

[1848] The drug was subsequently extracted using the respective dispensing systems and tested by a competitive inhibition assay. The test method has been developed from the literature (Velayudhan et al., Demonstration of functional similarity of proposed biosimilar ABP501 to adalimumab BioDrugs 30:339-351 (2016) and Barbeauet et al., Application Note: Screening for inhibitors of TNF/s TNFR1 Binding using AlphaScreen Technology. PerkinElmer Technical Note ASC-016. (2002)), as well as pre-testing development work using control drug and experiments using the provided AlphaLISA test kits. FIG. 68 demonstrates the principle of the competition assay performed in the experiment.

[1849] The bellows were loaded as follows: aseptically wiped the dispensing port of the simulated ingestible device jig with 70% ethanol; allowed to air dry for one minute; used an adalimumab delivery syringe to load each set of bellows with 200 L of drug; took a photo of the loaded device; gently rotated the device such that the drug is allowed to come in contact with all bellows surfaces; protected the bellows from light; and incubate at room temperature for the predetermined time period to allow full contact of the drug with all bellows' surfaces.

[1850] The drug was extracted as follows: after completion of the incubation period; the device jig was inverted such that the dispensing port was positioned over a sterile collection microfuge tube and petri dish below; five cubic centimeters of air was drawn into an appropriate syringe; the lure lock was attached to the device jig; the syringe was used to gently apply positive pressure to the bellow with air such that the drug was recovered in the collection microfuge tube; where possible, a video of drug dispensing was taken; samples were collected from each bellows type; a control drug sample was collected by directly dispensing 200 L of drug from the commercial dispensing syringe into a sterile microfuge tube; the control drug-free sample was collected by directly dispensing 200 L of PBS using a sterile pipette into a sterile microfuge tube; the collected drug was protected from light; and the drug was diluted over the following dilution range (250, 125, 25, 2.5, 0.25, 0.025, 0.0125, 0.0025 g) in sterile PBS to determine the IC.sub.50 range of the drug.

[1851] To determine any effects storage conditions may have on drug efficacy in the device, the drug (stored either in the syringe, silicon bellows, PVC bellows) was stored at room temperature while protected from light for 24 hours and 72 hours. Samples were then extracted and the steps in the preceding paragraph were repeated.

[1852] The AlphaLISA (LOCI) test method was used. Human TNF standard dilution ranges were prepared as described in Table 6.

TABLE-US-00042 TABLE 6 [human TNF] Vol. of in standard curve Vol. of diluent (g/mL (pg/mL Tube human TNF (L) (L) * in 5 L) in 5 L) A 10 L of reconstituted 90 1E07 100 000 human TNF B 60 L of tube A 140 3E08 30 000 C 60 L of tube B 120 1E08 10 000 D 60 L of tube C 140 3E09 3 000 E 60 L of tube D 120 1E09 1 000 F 60 L of tube E 140 3E10 300 G 60 L of tube F 120 1E10 100 H 60 L of tube G 140 3E11 30 I 60 L of tube H 120 1E11 10 J 60 L of tube I 140 3E12 3 K 60 L of tube J 120 1E12 1 L 60 L of tube K 140 3E13 0.3 M ** 0 100 0 0 (background) N ** 0 100 0 0 (background) O ** 0 100 0 0 (background) P ** 0 100 0 0 (background)

[1853] The test was performed as follows: the above standard dilution ranges were in a separate 96-well plate; to ensure consistent mixing, samples were mixed up and down gently with a pipette five times; a 384-well test plate was prepared according to the test layout diagram depicted Table 7; five microliters of 10,000 pg/mL TNF standard from the previously made dilution plate was added to each corresponding concentration as shown in Table 6; five microliters of recovered drug (directly from the commercial syringe (A), from the silicone bellows (B Si), from the PVC bellows (B PVC), or from the PBS control (C) was added into the corresponding wells described in Table 5; the test plate was incubated for one hour at room temperature while protected from light; 10 microliters of acceptor beads were added to each previously accessed well; the wells were incubated for 30 minutes at room temperature while protected from light; 10 L of biotinylated antibody was added to each previously accessed well; the wells were incubated for 15 minutes at room temperature, while protected from light; the room lights were darkened and 25 microliters of streptavidin (SA) donor beads were added to each previously accessed well; the wells were incubated for 30 minutes at room temperature while protected from light; the plate was read in Alpha Mode; and the results were recorded. Upon addition of reagent(s) in the various steps, each well was pipetted up and down three times to achieve good mixing.

TABLE-US-00043 TABLE 7 1 2 3 4 5 6 7 8 9 10 11 12 STD2 STD10 250 250 250 250 250 250 250 250 250 A 1.00E+05 10 A A A A A B Si B Si B Si B Si B C STD3 STD11 125 125 125 125 125 125 125 125 125 30000 3 A A A A A B Si B Si B Si B Si D STD4 STD12 25 25 25 25 25 25 25 25 25 E 10000 1 A A A A A B Si B Si B Si B Si F STD5 STD13 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 G 3000 0.333 A A A A A B Si B Si B Si B Si H STD6 Blank 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 I 1000 0 A A A A AB Si B Si B Si B Si J STD7 Blank 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 K 300 0 A A A A A B Si B Si B Si B Si L M STD8 Blank 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 100 0 A A A A A B Si B Si B Si B Si N O STD9 Blank 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 30 0 A A A A A B Si B Si B Si B Si P 13 14 15 16 17 18 19 20 21 22 23 250 250 250 250 250 250 250 250 250 250 250 A B Si B PVC B PVC B PVC B PVC B PVC C C C C C B C 125 125 125 125 125 125 125 125 125 125 125 B Si B PVC B PVC B PVC B PVC B PVC C C C C C D 25 25 25 25 25 25 25 25 25 25 25 E B Si B PVC B PVC B PVC B PVC B PVC C C C C C F 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 G B Si B PVC B PVC B PVC B PVC B PVC C C C C C H 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 I B Si B PVC B PVC B PVC B PVC B PVC C C C C C J 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 0.025 K B Si B PVC B PVC B PVC B PVC B PVC C C C C C L M 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 B Si B PVC B PVC B PVC B PVC B PVC C C C C C N O 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 0.003 B Si B PVC B PVC B PVC B PVC B PVC C C C C C P

[1854] The data are shown in FIGS. 69-71. The data demonstrate that the bellows do not negatively impact the drug function after shelf lives of 4 hours, 24 hours, or 336 hours. The IC.sub.50 values of the drug dispensed from the bellows were comparable to the IC.sub.50 values of the standard dispensation method (Table 6). A slight right shift was noted in the bellows curves after 24 hours (FIG. 70), but this shift was well within the error bars of the curves. Tables 8-11 represent data of FIGS. 69-71, respectively. Of note, when comparing mean (n=5) RFU data between test articles over the concentration ranges significant differences (p<0.05) were discerned. However, these significant differences did not favor either test article over time, suggesting that they were not related to the performance of the material in response to the drug (FIGS. 69-71).

TABLE-US-00044 TABLE 8 Needle Silicone PVC control (A) Bellows (B) Bellows (C) 4 Hours 0.0174 0.0169 0.0172 24 Hours 0.0180 0.0180 0.0180 336 Hours 0.0144 0.0159 0.0163

TABLE-US-00045 TABLE 9 Statistics (Student's T-test, 2 tailed, non- pair-wise, for significance p < 0.05) Drug Needle control (A) Needle control (A) Silicone (micrograms) vs. Silicone (B) vs. PVC vs. PVC 0.0001 0.911 0.008* 0.268 0.0025 0.138 0.390 0.822 0.0125 0.122 0.118 0.771 0.025 0.143 0.465 0.020* 0.25 0.591 0.984 0.350 2.5 0.243 0.124 0.169 125 0.867 0.688 0.182 250 0.681 0.184 0.108 *p < 0.5 data set

TABLE-US-00046 TABLE 10 Statistics (Student's T-test, 2 tailed, non- pair-wise, for significance p < 0.05) Drug Needle control (A) Needle control (A) Silicone (micrograms) vs. Silicone (B) vs. PVC vs. PVC 0.0001 0.132 0.038* 0.292 0.0025 0.003* 0.076 0.575 0.0125 0.161 0.022* 0.783 0.025 0.058 0.078 0.538 0.25 0.974 0.384 0.198 2.5 0.714 0.080 0.017* 125 0.873 0.731 0.269 250 0.798 0.956 0.903 *p < 0.5 data set

TABLE-US-00047 TABLE 11 Statistics (Student's T-test, 2 tailed, non- pair-wise, for significance p < 0.05) Drug Needle control (A) Needle control (A) Silicone (micrograms) vs. Silicone (B) vs. PVC vs. PVC 0.0001 0.858449 0.036847* 0.026444* 0.0025 0.087379 0.280302 0.046767* 0.0125 0.469282 0.057232 0.117194 0.025 0.02758* 0.078234 0.373419 0.25 0.411548 0.258928 0.400498 2.5 0.368959 0.156574 0.006719* 125 0.948649 0.246702 0.463735 250 0.485046 0.128993 0.705543 *p < 0.5 data set

Example 9. A Comparison Study of Systemic Vs Intracecal Delivery of SMAD7 Bio-Distribution in DSS-Induced Colitis in Male C57Bl/6 Mice

[1855] The objective of this study was to compare the efficacy of novel test articles, e.g., fluorescent SMAD7 antisense oligonucleotides (SMAD7 AS), when dosed systemically versus intracecally in the treatment of DSS-induced colitis, in male C57Bl/6 mice.

Experimental Design

[1856] A minimum of 10 days prior to the start of the experiment a cohort of animals underwent surgical implantation of a cecal cannula. A sufficient number of animals underwent implantation to allow for 12 cannulated animals to be enrolled in the main study (i.e., 16 animals).

[1857] Colitis was induced in 12 male C57Bl/6 mice (Groups 4-5) by exposure to 3% DSS-treated drinking water from Day 0 to Day 5. Three groups of six additional animals per group (n=6 cannulated; n=12 non-cannulated; Groups 1-3) served as no-disease controls (Groups 1-3). All animals were weighed daily and assessed visually for the presence of diarrhea and/or bloody stool during this time.

[1858] Animals were dosed with test-article via oral gavage (PO) or intracecal injection (IC) once on Day 9 as indicated in Table 12. The animals in Group 0 were not dosed. The animals in Groups 2 and 4 were dosed PO with SMAD7 antisense. The animals in Groups 3 and 5 were dosed IC with SMAD7 antisense.

[1859] All animals were euthanized by CO.sub.2 inhalation 12 hours after dosing, on Day 10. Terminal blood was collected into two K.sub.2EDTA tubes and processed for plasma. Both plasma and pellet samples were snap-frozen in liquid nitrogen and stored at 80 C. Cecum contents were removed and the contents were split into two aliquots. Both aliquots were weighed and snap frozen in separate cryovials in liquid nitrogen. The cecum was excised and bisected longitudinally; each piece is separately weighed and flash-frozen in liquid nitrogen. The colon contents were removed and the contents were split into two aliquots. Both aliquots were weighed and snap frozen in separate cryovials in liquid nitrogen. The colon was then rinsed, and the most proximal 2 cm of colon was collected. This 2-cm portion was bisected longitudinally; each piece was separately weighed and flash-frozen in liquid nitrogen. Snap-frozen blood pellet, cecum/colon contents, and tissue samples were used for downstream fluoremetry or RP-HPLC. The details of the study design are shown in Table 12.

TABLE-US-00048 TABLE 12 Study design Terminal No Cecal Colitis Collections Group Animals Cannula Induction Treatment Route Schedule Day 10 1 6 NO Whole blood, 2 6 NO Fluorescently PO QD plasma, cecal labeled Day 9** contents, colon SMAD7 contents, cecal 3 6 YES antisense IC tissue, colon tissue 4 6 NO 3% DSS 50 ng* PO Days 0-5 5 6 YES IC *Per mouse. TA is administered in 0.075 mL/animal. **Animals are dosed on Day 9 and collections are performed 12 hours later.

Materials and Methods

Mice

[1860] Normal male C57Bl/6 mice between the ages of 6-8 weeks old, weighing 20-24 g, were obtained from Charles River Laboratories. The mice were randomized into five groups of six mice each, and housed in groups of 8-15 per cage, and acclimatized for at least three days prior to entering the study. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour, with an automatic timer for a light/dark cycle of 12 hours on/off, and fed with Labdiet 5053 sterile rodent chow, with water administered ad libitum.

Cecal Cannulation

[1861] The animals were placed under isoflurane anesthesia, with the cecum exposed via a midline incision in the abdomen. A small point incision was made in the distal cecum, where 1-2 cm of the cannula was inserted. The incision was closed with a purse string suture using 5-0 silk. An incision was then made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was then washed copiously with warmed saline prior to closing the abdominal wall. A small incision was also made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All animals were administered 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until recovery before returning to their cage. All animals were administered 0.6 mg/kg BID buprenorphine for the first 3 days, and Baytril at 10 mg/Kg every day for the first 5 days post-surgery.

Disease Induction

[1862] Colitis was induced on Day 0 via addition of 3% DSS (MP Biomedicals, Cat #0260110) to the drinking water. Fresh DSS/water solutions was provided on Day 3 and any of the remaining original DSS solution is discarded.

Body Weight and Survival

[1863] Animals were observed daily (weight, morbidity, survival, presence of diarrhea and/or bloody stool) in order to assess possible differences among treatment groups and/or possible toxicity resulting from the treatments.

Animals Found Dead or Moribund

[1864] Animals were monitored on a daily basis. Animals exhibiting weight loss greater than 30% were euthanized, and samples were not collected from these animals.

Dosing

[1865] Animals were dosed with test-article via oral gavage (PO) or intracecal injection (IC) once on Day 9 as indicated in Table 12. Animals in Group 0 were not dosed. Animals in Groups 2 and 4 were dosed PO with SMAD7 antisense. Animals in Groups 3 and 5 were dosed IC with SMAD7 antisense.

Sacrifice

[1866] All animals were euthanized by CO.sub.2 inhalation 12 hours after dosing, on Day 10.

Sample Collection

[1867] Intestinal contents, peripheral blood and tissue were collected at sacrifice on Day 10, as follows:

Blood/Plasma

[1868] Terminal blood was collected into two K.sub.2EDTA tubes and processed for plasma. The approximate volume of each blood sample was recorded prior to centrifugation. Both plasma and pellet samples were snap-frozen in liquid nitrogen and stored at 80 C. The first pellet sample (sample 1) was used for fluoremetry. The second pellet sample (sample 2) was used for RP-HPLC.

Cecum Contents

[1869] Cecum contents was removed and contents were split into two aliquots. Both aliquots were weighed and snap frozen in separate cryovials in liquid nitrogen. The first sample (sample 1) was used for fluorometry. The second sample (sample 2) was used for RP-HPLC.

Cecum

[1870] The cecum was excised and bisected longitudinally; each piece was separately weighed and snap-frozen. The first sample (sample 1) was used for fluoremetry. The second sample (sample 2) was used for RP-HPLC.

Colon Contents

[1871] Colon contents were removed and contents were split into two aliquots. Both aliquots were weighed and snap frozen in separate cryovials in liquid nitrogen. The first sample (sample 1) was used for fluorometry. The second sample (sample 2) was used for RP-HPLC.

Colon

[1872] The colon was rinsed, and the most proximal 2 cm of colon was collected and bisected longitudinally. Each piece was separately weighed and flash-frozen in liquid nitrogen. The first sample (sample 1) was used for fluorometry. The second sample (sample 2) was used for RP-HPLC.

SMAD7 Antisense Bioanalysis

[1873] Samples flash-frozen for fluoremetry were homogenized in 0.5 mL buffer RLT+(Qiagen). Homogenate was centrifuged (4000g; 10 minutes), and supernatant was collected. Forty microliters of the sample was diluted 1:6 in 200 L of bicarbonate solution and 100 L of diluted supernatant was analyzed on a fluorescent plate reader (485 excitation; 535 emission) in duplicate.

[1874] Prior to the above, assay development was performed as follows. Samples (as indicated in Sample Collection) were harvested from a naive animal and flash-frozen. Samples were then homogenized in 0.5 mL buffer RLT+, homogenate was centrifuged (4000g; 10 minutes) and supernatant was collected and diluted 1:6 with bicarbonate solution (i.e., 0.5 mL supernatant was added to 2.5 mL of PBS). An aliquot (0.200 mL (90 L for each duplicate) of each diluted sample was pipetted into 15 (14 dilution of FAM-AS-SAMD7+ blank control) Eppendorf tubes. One tube was set-aside to be used as a blank sample. Ten microliters of fluorescently-labeled SMAD7 antisense was then spiked into all other sample to achieve final concentrations of 50 pg/mL, 16.67 pg/mL, 5.56 pg/mL, 1.85 pg/mL, 0.62 pg/mL, 0.21 pg/mL, 0.069 pg/mL, 0.023 pg/mL, 7.6 ng/mL, 2.5 ng/mL, 0.847 ng/mL, 0.282 ng/mL, 0.094 ng/mL, and 0.024 ng/mL respectively. The fluorescently-labeled SMAD7 antisense was prepared and serially diluted such that the volume added to each organ homogenate sample was the same for each of the above concentrations. These samples were analyzed on a fluorescent plate reader (485 excitation; 535 emission) in duplicate.

Processing for RP-HPLC

[1875] Samples flash-frozen for RP-HPLC were homogenized in buffer RLT+(Qiagen). Homogenate was centrifuged (4000g; 10 minutes), and supernatant was used to perform RP-HPLC analysis.

Results

[1876] The data in FIGS. 73 and 74 show that significantly more SMAD7 antisense oligonucleotide was present in cecum tissue and colon tissue for mice with or without DSS treatment that were intra-cecally administered the SMAD7 antisense oligonucleotide as compared to mice with or without DSS treatment that were orally administered the SMAD7 antisense oligonucleotide. The data in FIG. 75 show that there is about the same level of SMAD7 antisense oligonucleotide in the cecum contents of mice with or without DSS treatment that were orally or intra-cecally administered the SMAD7 antisense oligonucleotide. No SMAD7 antisense oligonucleotide was found in the plasma or white blood cell pellet of SMAD7 antisense oligonucleotide treated mice. No significant differences were observed in clinical observations, GI-specific adverse effects or toxicity due to FAM-AS-SMAD7 treatment via PO vs IC. No fluorescent detection of FAM-AS-SMAD7 was found in plasma and whole blood cell pellets across all treatment groups. A significant higher fluorescent signal (RFU) of FAM-AS-SMAD7 was found in cecum tissue when delivered intra-cecally compared with PO in both normal and DSS-induced models (FIG. 83). A slight higher RFU was also found in colon tissue when delivered intra-cecally, however, the overall signal is 10 times lower (FIG. 84). A significant higher RFU was found in colon content when delivered intra-cecally compared with PO in a normal mouse model (FIG. 85). This result was not seen in cecum content across all treatment groups (FIG. 86), indicating a better tissue absorption of oligos in cecum tissue from cecal content when delivered intra-cecally, but not in colon content at 12 hours post-treatment.

Example 10. Comparison of the Tissue, Plasma, and GI Content Pharmacokinetics of Tacrolimus Through Oral Vs. Intra-Cecal Ingestible Device Delivery in Yorkshire-Cross Farm Swine

[1877] The primary objective of this study was to compare the tissue, plasma, rectal sample, and GI content pharmacokinetics of tacrolimus through oral versus intra-cecal ingestible device delivery in normal Yorkshire-Cross farm swine.

[1878] This study compares the effects of administration of: a single intra-cecal administration of an ingestible device containing 0.8 mL sterile vehicle solution (80% alcohol, 20% castor oil (HCO-60)); a single oral dose of tacrolimus at 4 mg/0.8 mL (in sterile vehicle solution); and a single intra-cecal administration of an ingestible device containing either 1 mg/0.8 mL (in sterile vehicle solution), 2 mg/0.8 mL (in sterile vehicle solution), or 4 mg/0.8 mL (in sterile vehicle solution).

[1879] This study employed five groups of three female swine weighing approximately 45 to 50 kg at study start. Swine were randomly placed into animal rooms/pens as they are transferred from the delivery vehicle without regard to group. Group numbers were assigned to the rooms in order of room number. No further randomization procedure was employed. The study design is provided in Table 13.

TABLE-US-00049 TABLE 13 Study Design Table Group Days Pre-Dose Hours Post-dose General size Dose Route 11 10 5 1 1 0.5 1 2 3 4 6 12 Fast Food/Water ad libidum Observations clinical observations Day 10~5 body weight* & Day 1 Treatments (Groups) 1. Vehicle control n = 3 0.8 mL (20% IC HCO-60, 80% EtOH) Surgical placement of IC port** Euthanized (1 Ingestible n = 3 Device) 2. Tacrolimus (PO) n = 3 4 mg in 0.8 mL Oral Surgical placement of 0.08 mg/kg IC port** Euthanized (solution) n = 3 3. Tacrolimus (IC) n = 3 1 mg in 0.8 mL IC Surgical placement of 0.02 mg/kg IC port** Euthanized (1 Ingestible n = 3 Device) 4. Tacrolimus (IC) n = 3 2 mg in 0.8 mL IC Surgical placement of 0.04 mg/kg IC port** Euthanized (1 Ingestible n = 3 Device) 5. Tacrolimus (IC) n = 3 4 mg in 0.8 mL IC Surgical placement of 0.08 mg/kg IC port** Euthanized (1 Ingestible n = 3 Device) Tacrolimus (required) 20 mg Samples***** Plasma cephalic, jugular or catheter Rectal contents rectal Tissue*** x5 necropsy Luminal contents**** x5 necropsy Analysis (Agrilux Total Charles River) Samples Plasma [Tacrolimus] 105 15 15 15 15 15 15 15 Rectal contents [Tacrolimus] 60 15 15 15 15 Tissue (intact)*** [Tacrolimus] 105 105 Luminal contents [Tacrolimus] 75 75 Tissue after removing luminal content [Tacrolimus] 75 75 Notes: *Animal weight was ~45-50 kg for drug doses proposed. **Surgical placement of IC port in all animals to control. ***Tissue samples [drug] (five GI section cecum (CAC); proximal colon (PCN); transverse colon (TCN); distal colon (DCN); rectum (RTM), plus mesenteric lymph nodes and Peyer's Patch). ****Luminal contents (cecum (CAC); proximal colon (PCN); transverse colon (TCN); distal colon (DCN); rectum (RTM)).

[1880] Animals in Group 1 received an ingestible device containing 0.8 mL of vehicle solution (80% alcohol, 20% HCO-60). Animals in Group 2 received orally 4 mL liquid formulation of tacrolimus at 4 mg/0.8 mL per animal (Prograf: 5 mg/mL). Animals in Group 3 received intra-cecally an ingestible device containing tacrolimus at 1 mg in 0.8 mL per ingestible device. Animals in Group 4 received intra-cecally an ingestible device containing tacrolimus at 2 mg in 0.8 mL per ingestible device. Animals in Group 5 received intra-cecally an ingestible device containing tacrolimus at 4 mg in 0.8 mL per ingestible device. To control for potential confounding effects of the surgery, all groups fast on Day 11 at least 24 hr before being subjected to anesthesia followed by surgical placements of a cecal port by a veterinary surgeon at Day 10. All animals were fasted for at least 12 hr prior to dosing on Day 1. Animals were dosed via either intra-cecal dosing (IC) or oral dosing (PO) at Day 1 (between 6-8 p.m.). All animals resumed feeding at approximately 4 hours after dose (11-12 m. after dosing).

[1881] Animals in Group 1 (Vehicle Control) were administered a single intra-cecal ingestible device containing 0.8 mL Vehicle solution (80% alcohol, 20% castor oil (HCO-60) on Day 1. On Day 10 the animals were anesthetized, and a veterinary surgeon surgically placed an intra-cecal port in each animal. On Day 1, each animal was placed into a sling then a single intra-cecal ingestible device containing 0.8 mL vehicle solution (80% alcohol, 20% castor oil (HCO-60)) is introduced by the veterinary surgeon into the cecum via the cecal port in each animal. Following ingestible device placement, the animals were removed from the slings and placed back into their pens with water. All animals resumed feeding at approximately 4 hours after dose. Samples of rectal contents were collected for pharmacokinetic analyses from each animal at each of 1, 3, 6, and 12 hours post-ingestible device placement using a fecal swab (rectal swab). A total of 60 samples were collected.

[1882] Approximately 200400 mg of rectal content were collected, if available, with a fecal swab (Copan Diagnostics Nylon Flocked Dry Swabs, 502CS01). The fecal swab was pre-weighed and weighed after collection in the collection tube (Sterile Tube and Cap No Media, PFPM913S), and the sample weight was recorded. The fecal swab was broken via the breakpoint, and was stored in the collection tube, and immediately frozen at 70 C. Whole blood (2 mL) was collected into K.sub.2EDTA coated tubes for pharmacokinetics at each time-point of pre-dose and 1, 2, 3, 4, 6 and 12 hours post-dose. Immediately following euthanasia, tissue was collected. A total of 105 samples were collected.

[1883] For tissue necropsy, small intestine fluid and cecal fluid were collected separately from all the animals into two separate square plastic bottles, and stored at 20 C. The length and diameter of the cecum and the colon was measured from one animal in each group and recorded for reference. Tissues were collected for pharmacokinetic analyses and include mesenteric lymph nodes, a Peyer's Patch, and five gastrointestinal sections, including cecum, proximal colon, transverse colon, distal colon, and rectum. All samples were weighed, and the tissue sample weights were recorded. In each of the five gastrointestinal sections, tissue samples were collected in three different areas where the mucosal surface was visible and not covered by luminal content by using an 8.0-mm punch biopsy tool. Around 3 grams of the total punched sample were collected into a pre-weighed 15-mL conical tube, and the tissue weight was recorded. Three mesenteric lymph nodes were collected from different areas and weighed. At least one Peyer's Patch was collected and weighed. Tissues were snap-frozen in liquid nitrogen and stored frozen at approximately 70 C. or below (total of 105 samples).

[1884] Luminal contents were collected for pharmacokinetic analyses from the surface of the tissue from each of five gastrointestinal sections: cecum, proximal colon, transverse colon, distal colon, and rectum (total of 75). The contents were collected in pre-weighed 15-mL conical tubes and the sample weights were recorded. Samples were snap-frozen in liquid nitrogen stored frozen at approximately 70 C. or below.

[1885] After removing the luminal content, another set of tissue samples from 3 different areas were collected via an 8.0-mm punch biopsy in each section of the five tissue gastrointestinal sections described above. Around 3 grams of the total punched sample were collected into a pre-weighed 15-mL conical tube, and the tissue weight was recorded (total of 75). Tissues were snap-frozen in liquid nitrogen and stored frozen at approximately 70 C. or below.

[1886] A 30-cm length of jejunum (separated into two 15 cm lengths), and the remaining distal and transverse colon tissue sample (after tissue and luminal content were collected for PK) were collected in one animal in each group of treatment, snap-frozen in liquid nitrogen and stored frozen at approximately 70 C. or below. All samples for pharmacokinetic analyses were stored on dry ice before analyses.

[1887] Group 2 animals were administered a single oral dose of tacrolimus at 4 mg/0.8 mL (0.08-mg/kg) (in the vehicle solution) on Day 1. Plasma, rectal content sample, tissue collection, GI content collection and related procedures/storage/shipments was the same as those employed in Group 1.

[1888] Group 3 animals were administered a single intra-cecal ingestible device containing tacrolimus at 1-mg/0.8 mL (0.02 mg/kg) (in the vehicle solution) on Day 1 by a veterinary surgeon. Plasma, rectal content sample, tissue collection, GI content collection and related procedures/storage/shipments was the same as those employed in Group 1. All samples were analyzed for tacrolimus.

[1889] Group 4 animals were administered a single intra-cecal ingestible device of tacrolimus at 2 mg/0.8 mL (0.04 mg/kg) (in sterile vehicle solution) on Day 1 by a veterinary surgeon. Plasma, rectal content sample, tissue collection, GI content collection and related procedures/storage/shipments were the same as those employed in Group 1. All samples were analyzed for tacrolimus.

[1890] Group 5 animals are administered a single intra-cecal ingestible device containing tacrolimus at 4 mg/0.8 mL (0.08 mg/kg) (in the vehicle solution) on Day 1 by a veterinary surgeon. Plasma, rectal content sample, tissue collection, GI content collection and related procedures/storage/shipments were the same as those employed in Group 1. All samples were analyzed for tacrolimus.

[1891] Detailed clinical observations were conducted daily from Day 10 to 5, and on Day 1. Additional pen-side observations were conducted at least once each day. The animals remained under constant clinical observation for the entire 12 hours from dose until euthanasia. Body weights were collected on Day 10, Day 5, and pre-dose on Day 1. Animals were euthanized via injection of a veterinarian-approved euthanasia.

Test Article and Formulation

[1892] 1. Vehicle solution, 20 mL
Description: 80% alcohol, 20% PEG-60 castor oil
Physical characteristics: clear liquid solution.

2. Prograf (Tacrolimus Injection), 10 Ampules

[1893] Description: A sterile solution containing the equivalent of 5 mg anhydrous tacrolimus in 1 mL. Tacrolimus is macrolide immunosuppressant and the active ingredient of Prograf. 0.8 mL of Prograf (5 mg/mL) was administrated through oral gavage per animal in group 2. Prograf (5 mg/mL) was diluted 2 folds (2.5 mg/mL) and 4x folds (1.25 mg/mL) by using vehicle solution. 0.8 mL of each concentration, 1.25 mg/mL, 2.5 mg/mL, and 5 mg/mL of Prograf, was injected into a DSS ingestible device for group 3, 4, and 5.
Formulation: Each mL contained polyoxyl 60 hydrogenated castor oil (HCO-60), 200 mg, and dehydrated alcohol, USP, 80.0% v/v.
Physical characteristics: clear liquid solution.

3. DDS Ingestible Device Containing Tacrolimus

[1894] Description: Three (3) DDS ingestible devices containing vehicle solution for Group 1, three (3) DSS ingestible devices containing 1 mg tacrolimus for Group 3, three (3) DDS ingestible devices containing 2 mg tacrolimus for Group 4, and three (3) DDS ingestible devices containing 4 mg tacrolimus for Group 5.

Acclimation

[1895] Animals were acclimated prior to study initiation for at least 7 days. Animals in obvious poor health were not placed on study.

Concurrent Medication

[1896] Other than veterinary-approved anesthetics and medications used during surgery to install the ileocecal ports, or for vehicle or test article administration, and analgesia and antibiotics post-surgery, no further medications were employed.

Feed

[1897] All swine were fasted at least 24 hours before being anesthetized and properly medicated for surgery or overnight before dosing. Otherwise, animals were fed ad-libitum. Tap water was pressure-reduced and passed through a particulate filter, then a carbon filter prior to supply to an automatic watering system. Water was supplied ad libitum. There were no known contaminants in the feed or water that would be expected to interfere with this study.

Results

[1898] The data in FIG. 76 show that the mean concentration of tacrolimus in the cecum tissue and the proximate colon tissue were higher in swine that were inta-cecally administered tacrolimus as compared to swine that were orally administered tacrolimus. All blood trough concentrations were <10 ng/mL and exposure AUC<2000-12 ng.Math.h/mL (FIGS. 87-89). Significantly higher Cmax values (9.20 3.30 and 21.80 4.73 ng/mL) were observed in groups treated with high (0.09 mg/kg) and moderate (0.04 mg/kg) dose of tacrolimus when delivered through IC capsule as compared to the Cmax values following PO delivery of tacrolimus (0.09 mg/kg). Significantly higher tissue (spiral and transverse colon) and luminal content (spiral, transverse, and distal colon) concentrations were observed in groups treated with high and moderate dose tacrolimus delivered through IC capsule as compared to the levels observed in animals administered tacrolimus via PO. No measurable level of tracrolimus was detected in tissue when animals were delivered tacrolimus via PO, despite systemic concentrations equivalent to low dose IC group (0.02 mg/kg) (FIGS. 90 and 91). A higher rectal content concentration was observed at 12 hours post-treatment in the IC capsule groups (FIG. 92), while no detectable level was observed in the PO group.

These data suggest that intra-cecal administration of tacrolimus is able to locally deliver tacrolimus to the tissues in the GI tract of a mammal, while not decreasing the systemic immune system of a mammal.

Example 11. Comparison of the Tissue, Plasma, and GI Content Pharmacokinetics of Adalimumab Through SC Vs. Intra-Cecal Ingestible Device Delivery in Yorkshire-Cross Farm Swine in DSS-Induced Colitis

[1899] The purpose of this non-Good Laboratory Practice (GLP) study is to explore the PK/PD and bioavailability of adalimumab when applied to (Dextran Sulfate Sodium Salt) DSS-induced colitis in Yorkshire-cross farm swine, and to evaluate topical Humira (adalimumab or ADA) in DSS-colitis in swine. Colitis was induced in weanling YorkShire-Cross farm swine by administering DSS once daily for 7 consecutive days via oral gastric intubation. The dose levels were chosen based on the doses and regimens used to induce colitis in weanling pigs. The doses of DSS were 1.275 or 2.225 g/k/day for Groups 2 and 3 respectively.

[1900] This study used one group of 19- to 21-day old weanling swine, and 2 groups of three, 19- to 20-day old weanling swine that weighed from 6.5 to 7.5 kg on arrival. To induce colitis, on study day 1 through and including day 7, animals in Groups 2 and 3 were administered once daily oral (gastric intubation) doses of DSS at 8.5% or 15% w/v for dose levels of 1.275 or 2.25 g/kg/day, respectively (Groups 2 and 3 respectively, 2 hours before morning feeding). The Group 1 control animals were administered sterile saline only. Each animal was placed in a sling for dosing. Animals were fasted at least 6 hours prior to each dose. See Study Table below.

TABLE-US-00050 Total Animal DSS % mg/ Volume DSS ADA Group Route #.sup.1 w/v ml (ml) Total g.sup.2 g/kg Fequency.sup.3 needed treatment.sup.4 Endpoints.sup.5.6,7 1 oral/ 1 0 0 105 0 0 QD, 7 day 0 Day 8 Body (Animal gastric (Vehicle) weights, 1501) intubation clinical signs, & necropsy and IHC at 3 hr post ADA 2 oral/ 3 8.5% 85 105 8.925 1.275 QD, 7 day 187.425 Day 8 Body (Animals gastric (rectal 13 mg) weights, 2501, intubation clinical 2502, and signs, & 2504) necropsy and IHC at 3 hr post ADA 3 oral/ 3 15% 150 105 15.75 2.25 QD, 7 day 330.75 Day 8 Body gastric (rectal 13 mg) weights, intubation clinical signs, & necropsy and IHC at 3 hr post ADA 1. Animal weighed around 6.5-7.5 kg 2. Daily clinical signs and body weight were closely monitored throughout the study. If severe clinical signs or body weight loss is observed at day 1~3 after dosing, the DSS dosing was shortened to 5 days. 3. 0.8 mL of ADA solution was dosed rectally to the colon via an endoscope 4. Necropsy was done to observe GI inflammation and overall histopathology 5. 5-cm length opened tissue samples harvested for immunohistochemistry from terminal ileum, cecum, proximal colon; spiral colon, transverse colon; distal colon, rectum, and included other gastrointestinal sites of inflammation depending on the necropsy results. 6. ~3 g of punch biopsy sample and ~200 mg luminal content snap frozen for adalimumab measurement and three extra 5-cm length open tissue samples taking down for immunohistochemistry staining of ADA at the site where ADA was administrated. Additional tissue biopsy samples were collected from 3 different areas at proximal colon and proximal region of transverse colon in each animal.

[1901] The day following the last DSS dose, using endoscopy and a catheter, at 13 mg adalimumkab/0.8 mL/pig (one 40 mg adalimumab/0.8 mL dosage syringe was divided into 3 parts and diluted with PBS) was placed in the proximal portion of the descending colon just past the bend of the transverse colon. Alternatively, 13 mg of adalimumab was diluted with PBS to a volume suitable for dosing post-weanling swine. Prior to dosing, endoscopy photographs were taken of the mucosal surface of the colon. Animals were anesthetized during adalimumab dosing. Prior to adalimumab dosing, animals were housed on rubber mats to prevent ingestion of bedding material, and were fasted at least 24 hours. The colon was cleansed using an enema prior to the procedure.

[1902] All animals were properly euthanized approximately 3 hours post-adalimumab-dose for tissue collections and subjected to a gross necropsy with emphasis on the severity of colitis (immediately after euthanasia, in order to avoid autolytic changes). All samples for histology were fixed in a fixation medium and the punch-biopsy sample snap-frozen in liquid nitrogen and stored frozen (70 C.).

[1903] To measure drug content, tissue samples and luminal content were collected by gently removing and collecting luminal content first, then using an 8.0 mm-punch biopsy tool. Biopsies from three different areas at the site of adalimumab administration were collected in each animal. Additional tissue biopsy samples were collected from three different areas at the proximal colon, and the proximal region of transverse colon in each animal. Approximately 3 g of total punched sample and 200 mg of luminal content were collected in a pre-weighed conical tubes and the tissue weighed was recorded.

[1904] Approximately, a 5-cm length of open gastrointestinal tissue sample including terminal ileum, cecum (CAC); proximal colon (PCN); transverse colon (TCN); spiral colon, distal colon (DCN), and rectum was collected, gently rinsed in saline to remove luminal material, and individually fixed in fixation buffer (10% neutral buffered formalin). Also, a 5-cm length of open gastrointestinal tissue from 3 different areas near the site of adalimumab administration was collected and fixed in formalin in the same manner for immunohistochemical staining for adalimumab. Tissue samples for histopathology were fixed in 10% neutral buffered formalin for 18-24 hr, and transferred to 70% ethanol. HUMIRA was supplied in single-use, 1-mL pre-filled glass syringes, as a sterile, preservative-free solution for subcutaneous administration. The solution of HUMIRA was clear and colorless, with a pH of about 5.2. Each syringe delivered 0.8 mL (40 mg adalimumab) of drug product. Each vial contained approximately 0.9 mL of solution to deliver 0.8 mL (40 mg adalimumab) of drug product. Each 0.8 mL HUMIRA contained 40 mg adalimumab, 4.93 mg sodium chloride, 0.69 mg monobasic sodium phosphate dihydrate, 1.22 mg dibasic sodium phosphate dihydrate, 0.24 mg sodium citrate, 1.04 mg citric acid monohydrate, 9.6 mg mannitol, 0.8 mg polysorbate 80, and water for injection. Sodium hydroxide was added as necessary to adjust pH.

[1905] All animals were randomized into groups of three. Animals were dosed once with adalimumab via subcutaneous (SC), perirectal (PR), or intracecal (IC) administration.

[1906] The concentration of adalimumab and TNF was measured in plasma at 1, 2, 3, 4, 6, and 12 hours post-dose. The concentration of adalimumab was measured in rectal contents at 1, 3, 6, and 12 hours post-dose and in luminal content at 12 hours post-dose. Concentration of adalimumab and TNF, HER2, and total protein was measured in gastrointestinal tissue, e.g., cecum sample (CAC), proximal colon sample (PCN), transverse colon sample (TCN), distal colon sample (DCNi) inflamed, distal colon non-inflamed sample (DCNn), and rectum sample (RTM), at 12 hours post-dose.

[1907] Treatment with 8.5% DSS (oral; Day 1 to Day 7) induced mild body weight loss, hemorrhage diarrhea, soft bloody stool, and moderate colitis in swine. Necropsy revealed marked edema and full thickness of mucosal erosion from the proximal colon through the distal rectum. The 8.5% DSS-induced animals were treated with adalimumab at day 8. No significant differences in clinical observations, GI-specific adverse effects or toxicity due to adalimumab treatment were observed. The 15% DSS (oral; day 1 to day 7)-induced animals had marked mucosal sloughing and hemorrhage from cecum to rectum and severe colitis. All of the animals were euthanized early on day 5.

[1908] Significant lesions of colitis were found in animals treated with 8.5% DSS and were characterized by inflammation that involved mucosa and submucosa, loss of surface epithelium (erosion), and intestinal crypts (FIGS. 93 and 94). There was little, if any, evidence of regeneration. The ileum and cecum were unremarkable in all animals except cecum from one animal (animal 2504) that was treated with 8.5% DSS, which had lesions of inflammation and loss of surface and crypt epithelium (FIGS. 95-99). Lesions of colitis were significant and consistent in all other segments of the large intestine from animals treated with 8.5% DSS. The severity and character of the changes were not remarkably different among the different segments or among these animals. Staining for human IgG was most consistent and intense at the adalimumab administration site and localized to the luminal surface of the mucosal epithelium or inflammatory exudate at the luminal surface, and penetration of adalimumab is found in the lamina propria near the luminal surface (FIG. 100).

Example 12. Human Clinical Trial of Treatment of Ulcerative Colitis Using Adalimumab

[1909] As a proof of concept, the patient population of this study is patients that (1) have moderate to severe ulcerative colitis, regardless of extent, and (2) have had an insufficient response to a previous treatment, e.g., a conventional therapy (e.g., 5-ASA, corticosteroid, and/or immunosuppressant) or a FDA-approved treatment. In this placebo-controlled eight-week study, patients are randomized. All patient undergo a colonoscopy at the start of the study (baseline) and at week 8. Patients enrolled in the study are assessed for clinical status of disease by stool frequency, rectal bleeding, abdominal pain, physician's global assessment, and biomarker levels such as fecal calprotectin and hsCRP. The primary endpoint is a shift in endoscopy scores from Baseline to Week 8. Secondary and exploratory endpoints include safety and tolerability, change in rectal bleeding score, change in abdominal pain score, change in stool frequency, change in partial Mayo score, change in Mayo score, proportion of subjects achieving endoscopy remission, proportion of subjects achieving clinical remission, change in histology score, change in biomarkers of disease such as fecal calprotectin and hsCRP, level of adalimumab in the blood/tissue/stool, change in cytokine levels (e.g., TNF, IL-6) in the blood and tissue.

[1910] FIG. 72 describes an exemplary process of what would occur in clinical practice, and when, where, and how the ingestible device will be used. Briefly, a patient displays symptoms of ulcerative colitis, including but not limited to: diarrhea, bloody stool, abdominal pain, high c-reactive protein (CRP), and/or high fecal calprotectin. A patient may or may not have undergone a colonoscopy with diagnosis of ulcerative colitis at this time. The patient's primary care physician refers the patient. The patient undergoes a colonoscopy with a biopsy, CT scan, and/or MRI. Based on this testing, the patient is diagnosed with ulcerative colitis. Most patients are diagnosed with ulcerative colitis by colonoscopy with biopsy. The severity based on clinical symptoms and endoscopic appearance, and the extent, based on the area of involvement on colonoscopy with or without CT/MRI is documented. Treatment is determined based on diagnosis, severity and extent.

[1911] For example, treatment for a patient that is diagnosed with ulcerative colitis is an ingestible device programmed to release a single bolus of a therapeutic agent, e.g., 40 mg adalimumab, in the cecum or proximal to the cecum. Prior to administration of the treatment, the patient is fasted overnight and is allowed to drink clear fluids. Four hours after swallowing the ingestible device, the patient can resume a normal diet. An ingestible device is swallowed at the same time each day. The ingestible device is not recovered.

[1912] In some embodiments, there may be two different ingestible devices: one including an induction dose (first 8 to 12 weeks) and a different ingestible device including a different dose or a different dosing interval.

[1913] In some examples, the ingestible device can include a mapping tool, which can be used after 8 to 12 weeks of induction therapy, to assess the response status (e.g., based on one or more of the following: drug level, drug antibody level, biomarker level, and mucosal healing status). Depending on the response status determined by the mapping tool, a subject may continue to receive an induction regimen or maintenance regimen of adalimumab.

[1914] In different clinical studies, the patients may be diagnosed with Crohn's disease and the ingestible devices (including adalimumab) can be programmed to release adalimumab in the cecum, or in both the cecum and transverse colon.

[1915] In different clinical studies, the patients may be diagnosed with illeocolonic Crohn's disease and the ingestible devices (including adalimumab) can be programmed to release adalimumab in the late jejunum or in the jejunum and transverse colon.

Example 13. Pharmacokinetic Study of Oral Vs. Intra-Cecal Administration of Tacrolimus in Yorkshire-Cross Farm Swine

[1916] The primary objective of this study was to study the pharmacokinetics of oral versus intra-cecal administration of tacrolimus in normal Yorkshire-Cross farm swine.

[1917] This study compares the effects of administration of: a single intra-cecal administration of a device containing 0.8 mL sterile vehicle solution (80% alcohol, 20% castor oil (HCO-60)); a single oral dose of tacrolimus at 0.09 mg/kg (in sterile vehicle solution); and a single intra-cecal administration of a device containing either 0.02 mg/kg (in sterile vehicle solution), 0.04 mg/kg (in sterile vehicle solution), or 0.09 mg/kg (in sterile vehicle solution).

[1918] This study employed five groups of three female swine weighing approximately 45 to 50 kg at study start. Swine were randomly placed into animal rooms/pens as they are transferred from the delivery vehicle without regard to group. Group numbers were assigned to the rooms in order of room number. No further randomization procedure was employed. The study design is provided in Table 14.

TABLE-US-00051 TABLE 14 Study Design Dosage HED Treatments mg/kg mg Route Endpoints Group Vehicle n = 3 0 0 Intra-cecal [Tacrolimus] in 1 control capsule blood and rectal content at 1~12 Group Tacrolimus n = 3 0.09 6.60 Oral solution hr post dose, and 2 GI tissue & GI Group Tacrolimus n = 3 0.02 1.65 Intra-cecal content at 12 hr 3 capsule post dose Group Tacrolimus n = 3 0.04 3.30 Intra-cecal 4 capsule Group Tacrolimus n = 3 0.09 6.60 Intra-cecal 5 capsule

[1919] Animals in Group 1 received intra-cecally a device containing a vehicle solution (80% alcohol, 20% HCO-60). Animals in Group 2 received orally a liquid formulation of tacrolimus at 0.09 mg/kg per animal. Animals in Group 3 received intra-cecally a device containing tacrolimus at 0.02 mg/kg per device. Animals in Group 4 received intra-cecally a device containing tacrolimus 0.04 mg/kg per device. Animals in Group 5 received intra-cecally a device containing tacrolimus 0.09 mg/kg per device.

[1920] Samples of rectal contents were collected for pharmacokinetic analyses from each animal at each of 1, 3, 6, and 12 hours post-device placement using a fecal swab (rectal swab).

[1921] The concentration of tacrolimus measured was measured in the blood at 1-, 2-, 3-, 4-, 6-, and 12-hours post-dose. The concentration of tacrolimus was measured in rectal contents at 1-, 3-, 6-, and 12-hours post-dose, and in the gastrointestinal tissue and luminal content, e.g., the cecum tissue and lumen, the proximal colon tissue and lumen, the spiral colon tissue and lumen, the transverse colon tissue and lumen, and the distal colon tissue and lumen, at 12 hours post-dose.

Results

[1922] The data in FIGS. 77 and 78 show that the mean concentration and AUC.sub.0-12 hours of tacrolimus in the blood was higher in swine that were intra-cecally administered tacrolimus as compared to swine that were orally administered tacrolimus even at the same concentration (0.09 mg/kg). The data in FIG. 79 show that the mean concentration of tacrolimus in the spiral colon tissue and the transverse colon tissue were statistically higher in swine that were inta-cecally administered tacrolimus as compared to swine that were orally administered tacrolimus. The data in FIG. 80 show that the mean concentration of tacrolimus in the spiral colon lumen, the transverse colon lumen, and the distal colon lumen were statistically higher in swine that were inta-cecally administered tacrolimus as compared to swine that were orally administered tacrolimus. The data in FIGS. 81 and 82 show that the mean concentration of tacrolimus in the rectal concent was higher in swine that were intra-cecally administered tacrolimus as compared to swine that were orally administered tacrolimus even at the same concentration, particularly at 12 hours post-dose.

[1923] These data suggest that intra-cecal administration of tacrolimus is able to locally deliver tacrolimus to the tissues in the GI tract of a mammal.

[1924] A summary of the results are shown in Table 15.

TABLE-US-00052 TABLE 15 Summary of Results Route PO IC IC IC Dosage (mg/kg) 0.09 0.02 0.04 0.09 Cmax (ng/ml) 3.53 2.39 9.197 21.8 3.84 0.57 3.30 4.73 Trough (12 hr) 0.568 0.746 1.96 4.35 (ng/ml) 0.291 0.038 0.491 0.561 AUC.sub.0-12 hr 16.83 15.29 51.35 129.6 (ng .Math. hr/ml) 3.641 2.36 4.04 7.83

[1925] Tables 16 and 17 provide the tissue and plasma ratios of the animals in Groups 2-5.

TABLE-US-00053 TABLE 16-1 Tissue .sub.(mean) (ng/g)/AUG.sub.(0-12 hr)(ng.Math.hr/ml) ratios Group 2 PO (0.09 mg/kg) Group 3 IC (0.02 mg/kg) Tissue AUC 0-12 hr Tissue AUC 0-12 hr (ng/g) (ng.Math.hr/ml) Ratio (ng/g) (ng.Math.hr/ml) Ratio Cecum 16.83 0 15.29 0.00 Proximal 16.83 0 50.20 15.29 3.28 Colon Spiral colon 16.83 0 204.00 15.29 13.34 Transverse 16.83 0 128.20 15.29 8.38 colon Distal Colon 16.83 0 44.70 15.29 2.92 TABLE 16-2 Tissue .sub.(mean) (ng/g)/AUG.sub.(0-12 hr)(ng.Math.hr/ml) ratios Group 4 IC (0.04 mg/kg) Group 5 IC (0.09 mg/kg) Tissue AUC 0-12 hr Tissue AUC 0-12 hr (ng/g) (ng.Math.hr/ml) Ratio (ng/g) (ng.Math.hr/ml) Ratio Cecum 52.3 51.35 1.019 77.3 129.6 0.60 Proximal 98.3 51.35 1.914 157.0 129.6 1.21 Colon Spiral colon 342.3 51.35 6.667 783.3 129.6 6.04 Transverse 85.8 51.35 1.670 272.0 129.6 2.10 colon Distal Colon 28.7 51.35 0.559 67.7 129.6 0.52

TABLE-US-00054 TABLE 17-1 Tissue .sub.(mean) (ng/g)/Trough.sub.(12 hr)(ng/ml) Group 2 PO (0.09 mg/kg) Group 3 IC (0.02 mg/kg) Tissue Trough level Tissue Trough level (ng/g) (12 hr) Ratio (ng/g) (12 hr) Ratio Cecum 0.568 0 0.746 0.00 Proximal 0.568 0 50.20 0.746 67.29 Colon Spiral colon 0.568 0 204.00 0.746 273.46 Transverse 0.568 0 128.20 0.746 171.85 colon Distal Colon 0.568 0 44.70 0.746 59.92 TABLE 17-2 Tissue .sub.(mean) (ng/g)/Trough.sub.(12 hr)(ng.Math.hr/ml) Group 4 IC (0.04 mg/kg) Group 5 IC (0.09 mg/kg) Tissue Trough Tissue Trough (ng/g) level (12 hr) Ratio (ng/g) level (12 hr) Ratio Cecum 52.3 1.96 26.684 77.3 4.35 17.78 Proximal 98.3 1.96 50.136 157.0 4.35 36.09 Colon Spiral colon 342.3 1.96 174.660 783.3 4.35 180.08 Transverse 85.8 1.96 43.759 272.0 4.35 62.53 colon Distal Colon 28.7 1.96 14.643 67.7 4.35 15.56

Example 14

[1926] An ingestible medical device according to the disclosure (TLC1) was tested on 20 subjects to investigate its localization ability. TLC1 was a biocompatible polycarbonate ingestible device that contained a power supply, electronics and software. An onboard software algorithm used time, temperature and reflected light spectral data to determine the location of the ingestible device as it traveled the GI tract. The ingestible device is 0.511.22 inches which is larger than a vitamin pill which is 0.40.85 inches. The subjects fasted overnight before participating in the study. Computerized tomography (CT) were used as a basis for determining the accuracy of the localization data collected with TLC1. One of the 20 subjects did not follow the fasting rule. CT data was lacking for another one of the 20 subjects. Thus, these two subjects were excluded from further analysis. TLC1 sampled RGB data (radially transmitted) every 15 seconds for the first 14 hours after it entered the subject's stomach, and then samples every five minutes after that until battery dies. TLC1 did not start to record optical data until it reached the subject's stomach. Thus, there was no RGB-based data for the mouth-esophagus transition for any of the subjects.

[1927] In addition, a PillCam SB (Given Imaging) device was tested on 57 subjects. The subjects fasted overnight before joining the study. PillCam videos were recorded within each subject. The sampling frequency of PillCam is velocity dependent. The faster PillCam travels, the faster it would sample data. Each video is about seven to eight hours long, starting from when the ingestible device was administrated into the subject's mouth. RGB optical data were recorded in a table. A physician provided notes on where stomach-duodenum transition and ileum-cecum transition occurred in each video. Computerized tomography (CT) was used as a basis for determining the accuracy of the localization data collected with PillCam.

Esophagus-Stomach Transition

[1928] For TLC1, it was assumed that this transition occurred one minute after the patient ingested the device. For PillCam, the algorithm was as follows: [1929] 1. Start mouth-esophagus transition detection after ingestible device is activated/administrated [1930] 2. Check whether Green<102.3 and Blue<94.6 [1931] a. If yes, mark as mouth-esophagus transition [1932] b. If no, continue to scan the data [1933] 3. After detecting mouth-esophagus transition, continue to monitor Green and Blue signals for another 30 seconds, in case of location reversal [1934] a. If either Green>110.1 or Blue>105.5, mark it as mouth-esophagus location reversal [1935] b. Reset the mouth-esophagus flag and loop through step 2 and 3 until the confirmed mouth-esophagus transition detected [1936] 4. Add one minute to the confirmed mouth-esophagus transition and mark it as esophagus-stomach transition

[1937] For one of the PillCam subjects, there was not a clear cut difference between the esophagus and stomach, so this subject was excluded from future analysis of stomach localization. Among the 56 valid subjects, 54 of them have correct esophagus-stomach transition localization. The total agreement is 54/56=96%. Each of the two failed cases had prolonged esophageal of greater than one minute. Thus, adding one minute to mouth-esophagus transition was not enough to cover the transition in esophagus for these two subjects.

Stomach-Duodenum

[1938] For both TLC1 and PillCam, a sliding window analysis was used. The algorithm used a dumbbell shape two-sliding-window approach with a two-minute gap between the front (first) and back (second) windows. The two-minute gap was designed, at least in part, to skip the rapid transition from stomach to small intestine and capture the small intestine signal after ingestible device settles down in small intestine. The algorithm was as follows: [1939] 1. Start to check for stomach-duodenum transition after ingestible device enters stomach [1940] 2. Setup the two windows (front and back) [1941] a. Time length of each window: 3 minutes for TLC1; 30 seconds for PillCam [1942] b. Time gap between two windows: 2 minutes for both devices [1943] c. Window sliding step size: 0.5 minute for both devices [1944] 3. Compare signals in the two sliding windows [1945] a. If difference in mean is higher than 3 times the standard deviation of Green/Blue signal in the back window [1946] i. If this is the first time ever, record the mean and standard deviation of signals in the back window as stomach reference [1947] ii. If mean signal in the front window is higher than stomach reference signal by a certain threshold (0.3 for TLC1 and 0.18 for PillCam), mark this as a possible stomach-duodenum transition [1948] b. If a possible pyloric transition is detected, continue to scan for another 10 minutes in case of false positive flag [1949] i. If within this 10 minutes, location reversal is detected, the previous pyloric transition flag is a false positive flag. Clear the flag and continue to check [1950] ii. If no location reversal has been identified within 10 minutes following the possible pyloric transition flag, mark it as a confirmed pyloric transition [1951] c. Continue monitoring Green/Blue data for another 2 hours after the confirmed pyloric transition, in case of location reversal [1952] i. If a location reversal is identified, flag the timestamp when reversal happened and then repeat steps a-c to look for the next pyloric transition [1953] ii. If the ingestible device has not gone back to stomach 2 hours after previously confirmed pyloric transition, stops location reversal monitoring and assume the ingestible device would stay in intestinal area

[1954] For TLC1, one of the 18 subjects had too few samples (<3 minutes) taken in the stomach due to the delayed esophagus-stomach transition identification by previously developed localization algorithm. Thus, this subject was excluded from the stomach-duodenum transition algorithm test. For the rest of the TLC1 subjects, CT images confirmed that the detected pyloric transitions for all the subjects were located somewhere between stomach and jejunum. Two out of the 17 subjects showed that the ingestible device went back to stomach after first the first stomach-duodenum transition. The total agreement between the TLC1 algorithm detection and CT scans was 17/17=100%.

[1955] For one of the PillCam subjects, the ingestible device stayed in the subject's stomach all the time before the video ended. For another two of the PillCam subjects, too few samples were taken in the stomach to run the localization algorithm. These three PillCam subjects were excluded from the stomach-duodenum transition localization algorithm performance test. The performance summary of pyloric transition localization algorithm for PillCam was as follows: [1956] 1. Good cases (48 subjects): [1957] a. For 25 subjects, our detection matches exactly with the physician's notes [1958] b. For 19 subjects, the difference between the two detections is less than five minutes [1959] c. For four subjects, the difference between the two detections is less than 10 minutes (The full transition could take up to 10 minutes before the G/B signal settled) [1960] 2. Failed cases (6 subjects): [1961] a. Four subjects had high standard deviation of Green/Blue signal in the stomach [1962] b. One subject had bile in the stomach, which greatly affected Green/Blue in stomach [1963] c. One subject had no Green/Blue change at pyloric transition

[1964] The total agreement for the PillCam stomach-duodenum transition localization algorithm detection and physician's notes was 48/54=89%.

Duodenum-Jejunum Transition

[1965] For TLC1, it was assumed that the device left the duodenum and entered the jejunum three minutes after it was determined that the device entered the duodenum. Of the 17 subjects noted above with respect to the TLC1 investigation of the stomach-duodenum transition, 16 of the subjects mentioned had CT images that confirmed that the duodenum-jejunum transition was located somewhere between stomach and jejunum. One of the 17 subjects had a prolonged transit time in duodenum. The total agreement between algorithm detection and CT scans was 16/17=94%.

[1966] For PillCam, the duodenum-jejunum transition was not determined.

Jejunum-Ileum Transition

[1967] It is to be noted that the jejunum is redder and more vascular than ileum, and that the jejunum has a thicker intestine wall with more mesentery fat. These differences can cause various optical responses between jejunum and ileum, particularly for the reflected red light signal. For both TLC1 and PillCam, two different approaches were explored to track the change of red signal at the jejunum-ileum transition. The first approach was a single-sliding-window analysis, where the window is 10 minutes long, and the mean signal was compared with a threshold value while the window was moving along. The second approach was a two-sliding-window analysis, where each window was 10 minutes long with a 20 minute spacing between the two windows. The algorithm for the jejunum-ileum transition localization was as follows: [1968] 1. Obtain 20 minutes of Red signal after the duodenum-jejunum transition, average the data and record it as the jejunum reference signal [1969] 2. Start to check the jejunum-ileum transition 20 minutes after the device enters the jejunum [1970] a. Normalize the newly received data by the jejunum reference signal [1971] b. Two approaches: [1972] i. Single-sliding-window analysis [1973] Set the transition flag if the mean of reflected red signal is less than 0.8 [1974] ii. Two-sliding-window analysis: [1975] Set the transition flag if the mean difference in reflected red is higher than 2 the standard deviation of the reflected red signal in the front window

[1976] For TLC1, 16 of the 18 subjects had CT images that confirmed that the detected 20 jejunum-ileum transition fell between jejunum and cecum. The total agreement between algorithm and CT scans was 16/18=89%. This was true for both the single-sliding-window and double-sliding-window approaches, and the same two subjects failed in both approaches.

[1977] The performance summary of the jejunum-ileum transition detection for PillCam is listed below: [1978] 1. Single-sliding-window analysis: [1979] a. 11 cases having jejunum-ileum transition detected somewhere between jejunum and cecum [1980] b. 24 cases having jejunum-ileum transition detected after cecum [1981] c. 19 cases having no jejunum-ileum transition detected [1982] d. Total agreement: 11/54=20% [1983] 2. Two-sliding-window analysis: [1984] a. 30 cases having jejunum-ileum transition detected somewhere between jejunum and cecum [1985] b. 24 cases having jejunum-ileum transition detected after cecum [1986] c. Total agreement: 30/54=56%

Ileum-Cecum Transition

[1987] Data demonstrated that, for TLC1, mean signal of reflected red/green provided the most statistical difference before and after the ileum-cecum transition. Data also demonstrated that, for TLC1, the coefficient of variation of reflected green/blue provided the most statistical contrast at ileum-cecum transition. The analysis based on PillCam videos showed very similar statistical trends to those results obtained with TLC1 device. Thus, the algorithm utilized changes in mean value of reflected red/green and the coefficient of variation of reflected green/blue. The algorithm was as follows: [1988] 1. Start to monitor ileum-cecum transition after the ingestible device enters the stomach [1989] 2. Setup the two windows (front (first) and back (second)) [1990] a. Use a five-minute time length for each window [1991] b. Use a 10-minute gap between the two windows [1992] c. Use a one-minute window sliding step size [1993] 3. Compare signals in the two sliding windows [1994] a. Set ileum-cecum transition flag if [1995] i. Reflected red/green has a significant change or is lower than a threshold [1996] ii. Coefficient of variation of reflected green/blue is lower than a threshold [1997] b. If this is the first ileum-cecum transition detected, record average reflected red/green signal in small intestine as small intestine reference signal [1998] c. Mark location reversal (i.e. ingestible device returns to terminal ileum) if [1999] i. Reflected red/green is statistically comparable with small intestine reference signal [2000] ii. Coefficient of variation of reflected green/blue is higher than a threshold [2001] d. If a possible ileum-cecum transition is detected, continue to scan for another 10 minutes for TLC1 (15 minutes for PillCam) in case of false positive flag [2002] i. If within this time frame (10 minutes for TLC1, 15 minutes for PillCam), location reversal is detected, the previous ileum-cecum transition flag is a false positive flag. Clear the flag and continue to check [2003] ii. If no location reversal has been identified within this time frame (10 minutes for TLC1, 15 minutes for PillCam) following the possible ileum-cecum transition flag, mark it as a confirmed ileum-cecum transition [2004] e. Continue monitoring data for another 2 hours after the confirmed ileum-cecum transition, in case of location reversal [2005] i. If a location reversal is identified, flag the timestamp when reversal happened and then repeat steps a-d to look for the next ileum-cecum transition [2006] ii. If the ingestible device has not gone back to small intestine 2 hours after previously confirmed ileum-cecum transition, stop location reversal monitoring and assume the ingestible device would stay in large intestinal area

[2007] The flag setting and location reversal criteria particularly designed for TLC1 device were as follows: [2008] 1. Set ileum-cecum transition flag if [2009] a. The average reflected red/Green in the front window is less than 0.7 or mean difference between the two windows is higher than 0.6 [2010] b. And the coefficient of variation of reflected green/blue is less than 0.02 [2011] 2. Define as location reversal if [2012] a. The average reflected red/green in the front window is higher than small intestine reference signal [2013] b. And the coefficient of variation of reflected green/blue is higher than 0.086

[2014] For TLC1, 16 of the 18 subjects had CT images that confirmed that the detected ileum-cecum transition fell between terminal ileum and colon. The total agreement between algorithm and CT scans was 16/18=89%. Regarding those two subject where the ileum-cecum transition localization algorithm failed, for one subject the ileum-cecum transition was detected while TLC1 was still in the subject's terminal ileum, and for the other subject the ileum-cecum transition was detected when the device was in the colon.

[2015] Among the 57 available PillCam endoscopy videos, for three subjects the endoscopy video ended before PillCam reached cecum, and another two subjects had only very limited video data (less than five minutes) in the large intestine. These five subjects were excluded from ileum-cecum transition localization algorithm performance test. The performance summary of ileum-cecum transition detection for PillCam is listed below: [2016] 1. Good cases (39 subjects): [2017] a. For 31 subjects, the difference between the PillCam detection and the physician's notes was less than five minutes [2018] b. For 3 subjects, the difference between the PillCam detection and the physician's notes was less than 10 minutes [2019] c. For 5 subjects, the difference between the PillCam detection and the physician's notes was less than 20 minutes (the full transition can take up to 20 minutes before the signal settles) [2020] 2. Marginal/bad cases (13 subjects): [2021] a. Marginal cases (9 subjects) [2022] i. The PillCam ileum-cecum transition detection appeared in the terminal ileum or colon, but the difference between the two detections was within one hour [2023] b. Failed cases (4 subjects) [2024] i. Reasons of failure: [2025] 1. The signal already stabilized in the terminal ileum [2026] 2. The signal was highly variable from the entrance to exit [2027] 3. There was no statistically significant change in reflected red/green at ileum-cecum transition

[2028] The total agreement between ileocecal transition localization algorithm detection and the physician's notes is 39/52=75% if considering good cases only. Total agreement including possibly acceptable cases is 48/52=92.3%

Cecum-Colon Transition

[2029] Data demonstrated that, for TLC1, mean signal of reflected red/green provided the most statistical difference before and after the cecum-colon transition. Data also demonstrated that, for TLC1, the coefficient of variation of reflected blue provided the most statistical contrast at cecum-colon transition. The same signals were used for PillCam. The cecum-colon transition localization algorithm was as follows: [2030] 1. Obtain 10 minutes of reflected red/green and reflected blue signals after ileum-cecum transition, average the data and record it as the cecum reference signals [2031] 2. Start to check cecum-colon transition after ingestible device enters cecum (The cecum-colon transition algorithm is dependent on the ileum-cecum transition flag) [2032] a. Normalize the newly received data by the cecum reference signals [2033] b. Two-sliding-window analysis: [2034] i. Use two adjacent 10 minute windows [2035] ii. Set the transition flag if any of the following criteria were met [2036] The mean difference in reflected red/green was more than 4 the standard deviation of reflected red/green in the back (second) window [2037] The mean of reflected red/green in the front (first) window was higher than 1.03 [2038] The coefficient of variation of reflected blue signal in the front (first) window was greater than 0.23

[2039] The threshold values above were chosen based on a statistical analysis of data taken by TLC1.

[2040] For TLC1, 15 of the 18 subjects had the cecum-colon transition detected somewhere between cecum and colon. One of the subjects had the cecum-colon transition detected while TLC1 was still in cecum. The other two subjects had both wrong ileum-cecum transition detection and wrong cecum-colon transition detection. The total agreement between algorithm and CT scans was 15/18=83%.

[2041] For PillCam, for three subjects the endoscopy video ended before PillCam reached cecum, and for another two subjects there was very limited video data (less than five minutes) in the large intestine. These five subjects were excluded from cecum-colon transition localization algorithm performance test. The performance summary of cecum-colon transition detection for PillCam is listed below: [2042] 1. 27 cases had the cecum-colon transition detected somewhere between the cecum and the colon [2043] 2. one case had the cecum-colon transition detected in the ileum [2044] 3. 24 cases had no cecum-colon transition localized

[2045] The total agreement: 27/52=52%.

[2046] The following table summarizes the localization accuracy results.

TABLE-US-00055 Transition TLC1 PillCam Stomach-Duodenum 100% (17/17) 89% (48/54) Duodenum-Jejunum 94% (16/17) N/A Ileum-Cecum 89% (16/18) 75% (39/52) Ileum-terminal 100% (18/18) 92% (48/52) ileum/cecum/colon

Example 16. Intracecal Administration of Therapeutic Antibodies in a Colitis Animal Model that has Previously Received an Adoptive T-Cell Transfer

[2047] A set of experiments were performed to compare the efficacy of an anti-IL12 p40 antibody and an anti-TNF antibody when dosed systemically versus intracecally in the treatment of colitis induced through adoptive transfer of a subpopulation of CD44.sup./CD62L.sup.+ T cells isolated from C57BI/6 donor mice into RAG2.sup./ recipients.

Materials

Test System

[2048] Species/strain: Mice, C57Bl/6 (donors) and RAG2.sup./ (recipients; C57Bl/6 background) [2049] Physiological state: Normal/immunodeficient [2050] Age/weight range at start of study: 6-8 weeks (20-24 g) [2051] Animal supplier: Taconic [2052] Randomization: Mice were randomized into seven groups of 15 mice each, and two groups of eight mice each. [2053] Justification: T cells isolated from male C57Bl/6 wild type donors were transferred into male RAG2.sup./ recipient mice to induce colitis. [2054] Replacement: Animals were not replaced during the course of the study.

Animal Housing and Environment

[2055] Housing: Mice were housed in groups of 8-15 animals per cage prior to cannulation surgery. After cannulation surgery, cannulated animals were single-housed for seven days post-surgery. After this point, animals were again group-housed as described above. Non-cannulated animals (Group 9) were housed at 8 mice per cage. ALPHA-Dri bedding was used. Prior to colitis induction (i.e., during the cannulation surgeries), bedding was changed a minimum of once per week. After colitis induction, bedding was changed every two weeks, with of dirty cage material captured and transferred to the new cage. Additionally, bedding from Group 9 animals was used to supplement the bedding for all other groups at the time of cage change. [2056] Acclimation: Animals were acclimatized for a minimum of 7 days prior to study commencement. During this period, the animals were observed daily in order to reject animals that presented in poor condition. [2057] Environmental conditions: The study was performed in animal rooms provided with filtered air at a temperature of 70+/5 F. and 50%+/20% relative humidity. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour. The room was on an automatic timer for a light/dark cycle of 12 hours on and 12 hours off, with no twilight. [2058] Food/water and contaminants: Animals were maintained with Labdiet 5053 sterile rodent chow. Sterile water was provided ad libitum.

Test Articles

[2059] Test Article: IgG Control [2060] Name of the Test Article: InVivoMAb polyclonal rat IgG [2061] Source: BioXCell, catalog # BE0094 [2062] Storage conditions: 4 C. [2063] Vehicle: Sterile PBS [2064] Formulation Stability: Prepare fresh daily [2065] Dose: 0.625 mg/mouse; 0.110 mL/mouse IP and IC [2066] Frequency and duration of dosing: Days 0-49. 3/week (IPGroup 3); QD (ICGroup 4) [2067] Route and method of administration: IP or IC

Formulation:

[2068] For Group 3: On each day of dosing, dilute stock pAb to achieve 2.145 mL of a 5.68 mg/mL solution
For Group 4: On each day of dosing, dilute stock pAb to achieve 2.145 mL of a 5.68 mg/mL solution [2069] Test Article: Anti-IL12 p40 [2070] Name of the Test Article: InVivoMAb anti-mouse IL-12 p40 [2071] Source: BioXCell, catalog # BE0051 [2072] Storage conditions: 4 C. [2073] Vehicle: Sterile PBS [2074] Formulation Stability: Prepare fresh daily [2075] Dose: 0.625 mg/mouse (IP and IC); 0.110 mL/mouse IP and IC [2076] Frequency and duration of dosing: Days 0-49. 3/week (IPGroup 5); QD (ICGroup 6); [2077] Route and method of administration: IP or IC

Formulation:

[2078] For Group 5: On each dosing day, the stock mAb was diluted to achieve 1.716 mL of a 5.68 mg/mL solution.
For Group 6: On each dosing day, the stock mAb was diluted to achieve 1.716 mL of a 5.68 mg/mL solution. [2079] Test Article: anti-TNF [2080] Name of the Test Article: InVivoPlus anti-mouse TNF, clone XT3.11 [2081] Source: BioXCell, catalog # BP0058 [2082] Storage conditions: 4 C. [2083] Vehicle: Sterile PBS [2084] Formulation Stability: Prepare fresh daily [2085] Dose: 0.625 mg/mouse (IP and IC); 0.110 mL/mouse IP and IC [2086] Frequency and duration of dosing: Days 0-49. 3/week (IPGroup 7); QD (ICGroup 8); [2087] Route and method of administration: IP or IC

Formulation:

[2088] For Group 7:On each dosing day, the stock mAb was diluted to achieve 1.716 mL of a 5.68 mg/mL solution.
For Group 8: On each dosing day, the stock mAb was diluted to achieve 1.716 mL of a 5.68 mg/mL solution.

Methods

[2089] The details of the study design are summarized in Table 18. A detailed description of the methods used in this study is also provided below.

TABLE-US-00056 TABLE 18 Study Design Cell Blood No Cecal Transfer Schedule Collection Endpoints Group Animals Cannula (Day 0) Treatment Dose* Route (Days 0-42) (RO) Endoscopy (Day 42) 1 8 YES Day 13 Days 3 Hours 14, 28, 42 Post Dose: 2 15 0.5 10.sup.6 Vehicle IP; IC: IP: Colon nave (PBS; IP) 3/week weight/ T.sub.H Vehicle IC: QD Length, cells (PBS; IC) stool score 3 15 IgG Control 625 g IP: Terminal (IP) 3/week collection Vehicle IC: QD (all (PBS; IC) groups): 4 15 Vehicle 625 g IP: Cecal (PBS; IP) 3/week Contents, IgG Control IC: QD Colon (IC) Contents, 5 15 Anti-IL12p40 625 g IP: Plasma, (IP) 3/week small Vehicle IC: QD intestinal (PBS; IC) tissue, 6 15 Vehicle 625 g IP: colon (PBS; IP) 3/week tissue, Anti-IL12p40 IC: QD mLN, and (IC) 7 15 Anti-TNF 625 g IP: Peyers (IP) 3/week Patches Vehicle IC: QD (PBS; IC) 8 15 Vehicle 625 g IP: (PBS; IP) 3/week Anti-TNF (IC) IC: QD 9 8 NO

[2090] A minimum of 10-14 days prior to the start of the experiment a cohort of animals underwent surgical implantation of a cecal cannula. A sufficient number of animals underwent implantation to allow for enough cannulated animals to be enrolled in the main study. An additional n=8 animals (Group 9) served as no surgery/no disease controls.

[2091] Colitis was induced on Day 0 in male RAG2.sup./ mice by IP injection of 0.510.sup.6 CD44.sup./CD62L.sup.+ T cells isolated and purified from C57Bl/6 recipients. The donor cells were processed by first harvesting spleens from 80 C57Bl/6 mice and then isolating the CD44.sup./CD62L.sup.+ T cells using Miltenyi Magnetic-Activated Cell Sorting (MACS) columns. An additional eight mice (Group 1) served as no-disease controls, and eight mice (Group 9) served as no-cannulation and no-disease controls (sentinel animals for bedding). All recipient mice were weighed daily and assessed visually for the presence of diarrhea and/or bloody stool. The cages were changed every two weeks starting on Day 7, with care taken to capture of dirty cage material for transfer to the new cage. On Day 13, blood was collected via RO eye bleed, centrifuged, and plasma was aliquoted (50 L and remaining) and frozen for downstream analysis. The pelleted cells were re-suspended in buffer to determine the presence of T cells by FACS analysis of CD45.sup.+/CD4.sup.+ events.

[2092] Treatment with test article was initiated on Day 0 and was continued until Day 42 as outlined in Table 18. The animals in Groups 1 and 9 (n=8 per group; naive controls) were not treated with test article. The animals in Group 2 were treated IP with vehicle (PBS) 3/week and IC with vehicle QD. The animals in Group 3 were treated IP with IgG control 3/week and IC with vehicle (PBS) QD. The animals in Group 4 were treated IP with vehicle (PBS) 3/week and IC with IgG control QD. The animals in Group 5 were treated IP with anti-IL12 p40 antibody 3/week and IC with vehicle QD. The animals in Group 6 were treated IP with vehicle 3/week and IC with anti-IL12 p40 antibody QD. The animals in Group 7 were treated IP with anti-TNF antibody 3/week and IC with vehicle QD. The animals in Group 8 were treated IP with vehicle 3/week and IC with anti-TNF antibody QD.

[2093] The mice underwent HD video endoscopy on Days 14 (pre-dosing; baseline), 28, and 42 (before euthanasia) in order to assess colitis severity. Images were captured from each animal at the most severe region of disease identified during endoscopy. Additionally, stool consistency was scored during endoscopy using the parameters described herein. Following endoscopy on Day 42, the animals from all groups were sacrificed and terminal samples were collected.

[2094] The animals were euthanized by CO.sub.2 inhalation three hours after dosing on Day 42. Terminal blood samples were collected and plasma obtained from these samples. The resulting plasma was split into two separate cryotubes, with 50 L in one tube (Bioanalysis) and the remainder in a second tube (TBD). The cecum and colon contents were removed and the contents collected, weighed, and snap frozen in separate cryovials. The mesenteric lymph nodes were collected and flash-frozen in liquid nitrogen. The small intestine were excised and rinsed, and the most distal 2-cm of ileum was placed in formalin for 24 hours and then transferred to 70% ethanol for subsequent histological evaluation. The Peyer's patches were collected from the small intestine, and were flash-frozen in liquid nitrogen. The colon was rinsed, measured, weighed, and then trimmed to 6-cm in length and divided into 5 pieces as described in the above Examples. The most proximal 1-cm of colon was separately weighed, and flash-frozen for subsequent bioanalysis (PK) of test article levels. Of the remaining 5-cm of colon, the most distal and proximal 1.5-cm sections were each placed in formalin for 24 hours and then transferred to 70% ethanol for subsequent histological evaluation. The middle 2-cm portion was bisected longitudinally, and each piece was weighed, placed into two separate cryotubes, and snap frozen in liquid nitrogen; one of the samples was used for cytokine analysis and the other was used for myeloperoxidase (MPO) analysis. All plasma and frozen colon tissue samples were stored at 80 C. until used for endpoint analysis.

[2095] A more detailed description of the protocols used in this study are described below.

Cecal Cannulation

[2096] Animals were placed under isoflurane anesthesia, and the cecum was exposed via a mid-line incision in the abdomen. A small point incision was made in the distal cecum through which 1-2 cm of the cannula was inserted. The incision was closed with a purse-string suture using 5-0 silk. An incision was made in the left abdominal wall through which the distal end of the cannula was inserted and pushed subcutaneously to the dorsal aspect of the back. The site was washed copiously with warmed saline prior to closing the abdominal wall. A small incision was made in the skin of the back between the shoulder blades, exposing the tip of the cannula. The cannula was secured in place using suture, wound clips, and tissue glue. All of the animals received 1 mL of warm sterile saline (subcutaneous injection) and were monitored closely until fully recovered before returning to the cage. All animals received buprenorphine at 0.6 mg/kg BID for the first 3 days, and Baytril at 10 mg/Kg QD for the first 5 days following surgery.

Disease Induction

[2097] Colitis was induced on Day 0 in male RAG2.sup./ mice by IP injection (200 L) of 0.510.sup.6 CD44.sup./CD62L.sup.+ T cells (in PBS) isolated and purified from C57Bl/6 recipients.

Donor Cell Harvest

[2098] Whole spleens were excised from C57Bl/6 mice and immediately placed in ice-cold PBS. The spleens were dissociated to yield a single cell suspension and the red blood cells were lysed. The spleens were then processed for CD4.sup.+ enrichment prior to CD44.sup.CD62L.sup.+ sorting by MACS.

Dosing

[2099] Treatment with test article was initiated on Day 0 and continued until Day 42 as outlined in Table 18. The animals in Groups 1 and 9 (n=8 per group; naive control) were not treated with test article. The animals in Group 2 were treated IP with vehicle (PBS) 3/week and IC with vehicle QD. The animals in Group 3 were treated IP with IgG control 3/week and IC with vehicle (PBS) QD. The animals in Group 4 were treated IP with vehicle (PBS) 3/week and IC with IgG control QD. The animals in Group 5 were treated IP with anti-IL12 p40 antibody 3/week and IC with vehicle QD. The animals in Group 6 were treated IP with vehicle 3/week and IC with anti-IL12 p40 antibody QD. The animals in Group 7 were treated IP with anti-TNF antibody 3/week and IC with vehicle QD. The animals in Group 8 were treated IP with vehicle 3/week and IC with anti-TNF antibody QD.

Body Weight and Survival

[2100] The animals were observed daily (weight, morbidity, survival, presence of diarrhea and/or bloody stool) in order to assess possible differences among treatment groups and/or possible toxicity resulting from the treatments.

Animals Found Dead or Moribund

[2101] The animals were monitored on a daily basis and those exhibiting weight loss greater than 30% were euthanized, and did not have samples collected.

Endoscopy

[2102] Each mouse underwent video endoscopy on Days 14 (pre-dosing; baseline), 28, and 42 (before euthanasia) using a small animal endoscope (Karl Storz Endoskope, Germany), under isoflurane anesthesia. During each endoscopic procedure, still images as well as video were recorded to evaluate the extent of colitis and the response to treatment. Additionally, an image from each animal at the most severe region of disease identified during endoscopy was captured. Colitis severity was scored using a 0-4 scale (0=normal; 1=loss of vascularity; 2=loss of vascularity and friability; 3=friability and erosions; 4=ulcerations and bleeding). Additionally, stool consistency was scored during endoscopy using the scoring system described herein.

Sacrifice

[2103] All animals were euthanized by CO.sub.2 inhalation following endoscopy on Day 42 and three hours after test-article dosing.

Sample Collection

[2104] Terminal blood (plasma and cell pellet), Peyer's patches (Groups 1-8 only), small intestine and colon mLN (Groups 1-8 only), cecum contents, colon contents, small intestine, and colon were collected at euthanasia, as follows.

Blood

[2105] Terminal blood was collected by cardiac puncture and plasma generated from these samples. The resulting plasma was split into two separate cryotubes with 50 L in one tube (Bioanalysis), and the remainder in a second tube (TBD).

Mesenteric Lymph Nodes

[2106] The mesenteric lymph nodes were collected, weighed, snap-frozen in liquid nitrogen, and stored at 80 C.

Small Intestine

[2107] The small intestine was excised and rinsed, and the most distal 2-cm of ileum will be placed in formalin for 24 hours and then transferred to 70% ethanol for subsequent histological evaluation.

Peyer's Patches

[2108] The Peyer's patches were collected from the small intestine. The collected Peyer's patches were weighed, snap-frozen in liquid nitrogen, and stored at 80 C.

Cecum/Colon Contents

[2109] The cecum and colon were removed from each animal and contents collected, weighed, and snap-frozen in separate cryovials.

Colon

[2110] Each colon was rinsed, measured, weighed, and then trimmed to 6-cm in length and divided into 5 pieces as outlined herein. The most proximal 1-cm of colon was separately weighed, and snap frozen for subsequent bioanalysis (PK) of test article levels. Of the remaining 5-cm of colon, the most distal and proximal 1.5-cm sections were placed in formalin for 24 hours and then transferred to 70% ethanol for subsequent histological evaluation. The middle 2-cm portion was bisected longitudinally, and each piece weighed, placed into two separate cryotubes, and snap-frozen in liquid nitrogen; one of these samples was used for cytokine analysis and the other sample was used for MPO analysis.

Cytokine Levels in Colon Tissue

[2111] Cytokine levels (IFN, IL-2, IL-4, IL-5, IL-1, IL-6, IL-12 p40, and TNF) were assessed in colon tissue homogenate (all groups) by multiplex analysis. MPO levels were assessed by ELISA in colon tissue homogenate (all groups).

Results

[2112] The Disease Activity Index was determined in each mouse using a total score from the scoring system depicted below.

TABLE-US-00057 Disease Activity Index Description Score Colitis Severity Normal 0 loss of vascularity 1 loss of vascularity and friability 2 friability and erosions 3 ulcerations and bleeding 4 Stool Consistency Normal 0 Loose stool, soft, staying in shape 1 abnormal form with excess moisture 2 Watery or diarrhea 3 Bloody diarrhea 4 Body Weight Loss (%) X < 0% or gain weight 0 2% X < 5% 1 5% X < 10% 2 10% X < 15% 3 15% X < 20% 4 20% X < 25% 5 25% X < 30% 6 X 35% 7 Total Score 15

[2113] The data in FIG. 103 show that mice intracecally administered anti-TNF antibody (Group 8) had decreased disease activity index (DAI) as compared to mice intraperitoneally administered anti-TNF antibody (Group 7) at Day 42 of the study. The data in FIG. 104 show that mice intracecally administered anti-TNF antibody (Group 8) had decreased levels of TNF, IL-17A, and IL-4 in colonic tissue as compared to the levels in colonic tissue intraperitoneally administered anti-TNF antibody (Group 7), when assessed at Day 42 of the study. The data in FIG. 105 show that mice intracecally administered anti-IL12 p40 antibody (Group 6) had decreased disease activity index (DAI) as compared to mice intraperitoneally administered anti-L12 p40 antibody (Group 5) at Day 28 and Day 42 of the study. The data in FIG. 106 show that mice intracecally administered anti-IL12 p40 antibody (Group 6) had decreased levels of IFN-gamma, IL-6, IL-17A, TNF, IL-22, and IL-1b in colonic tissue as compared to the levels in colonic tissue in vehicle-administered control mice (Group 2).

Exemplary Embodiments

[2114] 1. A method of treating an an inflammatory disease or condition that arises in a tissue originating from the endoderm in a subject, comprising:

[2115] administering to the subject a pharmaceutical formulation that comprises an immune modulator,

wherein the pharmaceutical formulation is released at a location in the gastrointestinal tract of the subject. [2116] 2. The method of embodiment 1, wherein the pharmaceutical formulation is administered in an ingestible device. [2117] 3. The method of embodiment 1, wherein the pharmaceutical formulation is released from an ingestible device. [2118] 4. The method of embodiment 2 or 3, wherein the ingestible device comprises a housing, a reservoir containing the pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device, wherein the reservoir is releasably or permanently attached to the exterior of the housing or internal to the housing. [2119] 5. The method of embodiment 2 or 3, wherein the ingestible device comprises a housing, a reservoir containing the pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device, wherein the reservoir is internal to the device. [2120] 6. A method of treating a disease of the gastrointestinal tract in a subject, comprising:

[2121] administering to the subject an ingestible device comprising a housing, a reservoir containing a pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device;

[2122] wherein the reservoir is releasably or permanently attached to the exterior of the housing or internal to the housing;

[2123] wherein the pharmaceutical formulation comprises an immune modulator, and

[2124] the ingestible device releases the pharmaceutical formulation at a location in the gastrointestinal tract of the subject that is proximate to one or more sites of disease. [2125] 7. A method of treating a disease of the gastrointestinal tract in a subject, comprising:

[2126] administering to the subject an ingestible device comprising a housing, a reservoir containing a pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device;

[2127] wherein the reservoir is internal to the device;

[2128] wherein the pharmaceutical formulation comprises an immune modulator, and

[2129] the ingestible device releases the pharmaceutical formulation at a location in the gastrointestinal tract of the subject that is proximate to one or more sites of disease. [2130] 8. The method of any one of embodiments 4 to 7, wherein the housing is non-biodegradable in the GI tract. [2131] 9. The method of any one of embodiments 2 to 8, wherein the release of the formulation is triggered autonomously. [2132] 10. The method of any one of embodiments 2 to 9, wherein the device is programmed to release the formulation with one or more release profiles that may be the same or different at one or more locations in the GI tract. [2133] 11. The method of any one of embodiments 2 to 10, wherein the device is programmed to release the formulation at a location proximate to one or more sites of disease. [2134] 12. The method of embodiment 11, wherein the location of one or more sites of disease is predetermined. [2135] 13. The method of any one of embodiments 4 to 12, wherein the reservoir is made of a material that allows the formulation to leave the reservoir [2136] 14. The method of embodiment 13, wherein the material is a biodegradable material. [2137] 15. The method of any one of embodiments 2 to 14, wherein the release of the formulation is triggered by a pre-programmed algorithm. [2138] 16. The method of any one of embodiments 2 to 15, wherein the release of the formulation is triggered by data from a sensor or detector to identify the location of the device. [2139] 17. The method of embodiment 16, wherein the data is not based solely on a physiological parameter. [2140] 18. The method of any one of embodiments 2 to 17, wherein the device comprises a detector configured to detect light reflectance from an environment external to the housing. [2141] 19. The method of embodiment 18, wherein the release is triggered autonomously or based on the detected reflectance. [2142] 20. The method of any one of embodiments 2 to 19, wherein the device releases the formulation at substantially the same time as one or more sites of disease are detected. [2143] 21. The method of any one of embodiments 4 to 20, wherein the release mechanism is an actuation system. [2144] 22. The method of embodiment 21, wherein the actuation system is a chemical actuation system. [2145] 23. The method of embodiment 21, wherein the actuation system is a mechanical actuation system. [2146] 24. The method of embodiment 21, wherein the actuation system is an electrical actuation system. [2147] 25. The method of embodiment 21, wherein the actuation system comprises a pump and releasing the formulation comprises pumping the formulation out of the reservoir. [2148] 26. The method of embodiment 21, wherein the actuation system comprises a gas generating cell. [2149] 27. The method of any one of embodiments 2 to 26, wherein the device comprises an anchoring mechanism. [2150] 28. The method of any one of embodiments 1 to 27, wherein the formulation comprises a therapeutically effective amount of the immune modulator. [2151] 29. The method of any one of the preceding embodiments, wherein the formulation comprises a human equivalent dose (HED) of the immune modulator