Use of Immune Suppressive Domains as Medicaments

Abstract

The present invention concerns uses of immune suppressive domains. In particular, the present invention concerns a use of an immune suppressive domain (ISD) for immune suppression and for reduction of inflammation.

Claims

1. Pharmaceutical product comprising an immune suppressive domain from a virus fusion protein as an active substance, wherein said immune suppressive domain is from a virus of the Orthomyxoviridae family.

2. The pharmaceutical composition according to claim 1, wherein said pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier, preservative, buffer, and/or surfactant.

3. The pharmaceutical composition according to claim 1, wherein said immune suppressive domain is from a virus of the Influenza virus A genus.

4. The pharmaceutical composition according to claim 1, wherein said immune suppressive domain comprises SEQ ID NO: 4 and/or SEQ ID NO: 287.

5. The pharmaceutical composition according to claim 1, wherein said immune suppressive domain is connected to at least one additional immune suppressive domain to form a dimer.

6. The pharmaceutical composition according to claim 5, wherein said dimer is a homodimer and comprises at least two immune suppressive domains with SEQ ID NO. 4, wherein said immune suppressive domains are cross-linked by a disulfide bond at the N-terminal or the C-terminal.

7. A pharmaceutical composition comprising an immunosuppressive polypeptide that is a dimer formed by two peptides comprising SEQ ID NO: 4 as an active substance subject to at least one of the following provisos: (i) said pharmaceutical composition is a parenteral, subcutaneous or oral formulation; (ii) said pharmaceutical composition is an injection liquid or infusion liquid; (iii) said pharmaceutical composition comprises a pharmaceutically acceptable carrier selected from solid, semisolid and liquid; (iv) said pharmaceutical composition is a suspension, solution, emulsion, gel, cream, or powder; (v) said pharmaceutical composition comprises 0.5-25% by weight of said peptides in solution; or (vi) said two peptides comprise SEQ ID NO: 4 and an additional cysteine at the C-terminal or N-terminal end of SEQ ID NO: 4, wherein said cysteine residues crosslink the two peptides via a disulfide bond.

8. The pharmaceutical composition according to claim 7, wherein said pharmaceutical composition is selected from a parenteral composition, a subcutaneous formulation, and an oral composition.

9. The pharmaceutical composition according to claim 7, wherein said pharmaceutical composition is suitable for IV, oral, parenteral, intraperitoneal, subcutaneous or intramuscular administration.

10. A pharmaceutical composition according to claim 7, wherein said pharmaceutical composition is packaged in unit-dose or multi-dose sealed containers, and/or packaged in ampoules and/or vials.

11. The pharmaceutical composition according to claim 7, wherein said pharmaceutical composition is a vaccine.

12. The pharmaceutical composition according to claim 7 obtainable by freeze-drying or which is suitable to be stored in a freeze-dried (lyophilized) condition.

13. The pharmaceutical composition according to claim 7 comprising a pharmaceutically acceptable carrier, preservative, buffer, and/or surfactant.

14. A pharmaceutical composition according to claim 7, wherein one or more amino acid residues in said polypeptide are modified by in vitro or in vivo chemical derivatization, wherein said chemical derivatization is selected from acetylation, carboxylation, glycosylation and phosphorylation, or wherein said polypeptide comprises blocking groups and wherein said blocking groups are either coupled to the N-terminus or used in place of the N-terminal amino acid residue of said polypeptide and/or coupled to the C-terminus or used in place of the C-terminal amino acid residue of said polypeptide.

15. The pharmaceutical composition according to claim 7, wherein the two peptides comprise a cysteine residue at a C-terminal end of SEQ ID NO. 4.

16. The pharmaceutical composition according to claim 7, wherein each of the two peptides consists of SEQ ID NO. 287.

17. The pharmaceutical composition according to claim 7, wherein said immune suppressive domains are cross-linked by a disulfide bond at the N-terminal or the C-terminal.

18. The pharmaceutical composition according to claim 7, wherein said dimer is a homodimer.

19. The pharmaceutical composition of claim 7, wherein said composition contains from 0.5 to 25% by weight of the active substance in solution.

20. A peptide comprising SEQ ID NO. 287.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0080] INF ISD peptide is identical to INF-F#2 and are dimeric form of the peptide with the sequence [Seq id 287] GLFGAIAGFIENGWEGCGGEKEKEK

[0081] FIG. 1 shows the effect of the INF ISD peptide on TNF-alpha mRNA levels.

[0082] FIG. 2 shows the effect of peptide incubation on secreted TNF-alpha levels in the supernatant of THP-1 cells.

[0083] FIG. 3 shows the effect of the INF ISD peptide on IL-1 beta mRNA levels in THP-1 cells.

[0084] FIG. 4 shows the effect of the INF ISD peptide on IL-6 mRNA levels in THP-1 cells.

[0085] FIG. 5 shows the effect of the INF ISD peptide on IL-8 mRNA levels in THP-1 cells.

[0086] FIG. 6 shows the effect of the INF ISD peptide on IL-10 mRNA levels in THP-1 cells.

[0087] FIG. 7 shows the effect of the INF ISD peptide on NF-kappa B mRNA levels in THP-1 cells.

[0088] FIG. 8 shows the effect of the INF ISD peptide on the house holding gene RPL13a mRNA levels in THP-1 cells.

[0089] FIG. 9 shows that in the presence of INF F#2, cells treated with LPS release significantly lower amounts of cytokines.

[0090] BM DCs were treated with LPS for 16 hours. Cell supernatants were then collected and analyzed for type I IFN using bioassay. Before LPS treatment cells were either pretreated with INF F#2, with the deletion mutant DI6 or not pretreated with any peptide.

[0091] FIG. 10 shows inflammation-related enzyme and transcription factor gene expression kinetics of THP-1 monocytes stimulated with 1 .Math.g/ml LPS. Gene expression was expressed as relative gene expression towards RPL13a-expression and non-stimulated cells at time zero (ΔΔCt). Data shown are means + standard deviation from two independent biological replications.

[0092] FIG. 11 shows effects of INF ISD peptide on expression of NF-kappaB mRNA in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M, 60 .Math.M INF ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the medians ± standard deviation from two independent biological replications.

[0093] FIG. 12 shows effects of INF ISD peptide on expression of SP-1 mRNA in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M, 60 .Math.M INF ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the medians ± standard deviation from two independent biological replications.

[0094] FIG. 13 shows effects of INF ISD peptide on protein secretion of IL-8 in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M or 60 .Math.M INF ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the median ± standard deviation from three independent experiments performed in duplicates.

[0095] FIG. 14 shows effects of INF ISD peptide on protein secretion of IL-10 in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M or 60 .Math.M INF ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the median ± standard deviation from three independent experiments performed in duplicates.

[0096] FIG. 15 shows effect of different stimulus on the secretion of IFN-gamma in PBMCs. PBMCs were incubated either with 1 .Math.g/ml or 50 ng/ml PMA and 1 .Math.g/ml ionomycin or 10 ng/ml SEB for indicated time periods. Data shown are the medians ± standard deviation from three independent technical replicates.

[0097] FIG. 16 shows expression kinetics of IFN gamma expression in response to PMA/ionomycin treatment. Gene expression was expressed as relative gene expression towards RPL13a expression and non-stimulated cells at time zero (ΔΔ Ct). Data shown are the medians ± standard deviation from three independent technical replicates.

[0098] FIG. 17 shows effect of INF ISD on secretion of protein of IFN-gamma in PMA/ionomycin stimulated PBMCs. PBMCs were incubated with either medium alone, 30 .Math.M or 60 .Math.M Flu ISU or 30 .Math.M or 60 .Math.M control peptide, and stimulated with 50 ng/ml PMA and 1 .Math.g/ml ionomycin. Data shown are the medians ± standard deviation from three independent experiments performed in duplicates.

[0099] FIG. 18 shows effects of SARS ([Seq id 285] AEVQIDRLITGRLQSLQTYVCGGEKEKEK) or Filo ISD ([Seq id 286] GAAIGLAWIPYFGPAAECGGEKEKEK) on expression of TNF-alpha mRNA in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M, 60 .Math.M SARS or Filo ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the medians ± standard deviation from two independent biological replications.

[0100] FIG. 19 shows effects of SARS or Filo ISD on expression of IL-1 j3 mRNA in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M, 60 .Math.M SARS or Filo ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the medians ± standard deviation from two independent biological replications.

[0101] FIG. 20 shows effects of SARS or Filo ISD on expression of IL-1 β mRNA in LPS-stimulated THP-1 cells. THP-1 cells were incubated with either medium alone, 30 .Math.M, 60 .Math.M SARS or Filo ISD peptide or 30 .Math.M, 60 .Math.M control peptide, and stimulated with 1 .Math.g/ml LPS. Data shown are the medians ± standard deviation from two independent biological replications.

[0102] FIG. 21 shows interactions between INF ISD peptide (pFlu) and STING depends on distinct STING domains. To investigate further the interaction between STING and INF ISD peptide (pFlu) the C-terminal domian of STING was expressed with a HA-tag in HEK293 cells. STING was either in a wt form or with deletions. Lysates from tansfected cells were used for pulldown using biotinylated INF ISD peptide (pFlu) and streptavidin coated beads. The bead eluate was then immunoblotted using antibodies against HA-tag. As seen in the figure wt STING and the deletion mutant DN5 (162-N) was readily pulled down using INF ISD peptide (pFLu) whereas the deletion mutants DN6 (172-N) was not. These data indicate that amino acids 162-172 are necessary for interactions between pFlu and STING.

DETAILED DISCLOSURE

[0103] According to an embodiment, the present invention concerns compositions of one or more immunosuppressive peptides. Immunosuppressive polypeptides are polypeptides that are capable of suppressing an immune response in animals, including human beings and other animal such as domestic or agricultural (cats, dogs, cows, sheep, horses, pigs, etc.) or test species such as mouse, rats, rabbits and the like.

[0104] In one embodiment of the present invention the immunosuppressive polypeptides are capable of at least 5% inhibition of T-lymphocyte proliferation, at least 10%, at least 20%, such as at least 30%, at least 40%, at least 50%, such as at least 60%, such as at least 70% inhibition of T-lymphocyte proliferation. In particular embodiments the immunosuppressive peptides of the present invention are capable of at least 75% inhibition of T-lymphocyte proliferation, at least 80%, such as at least 85%, at least 90%, such as at least 95%, at least 97%, such as at least 99%, at least 100% inhibition of T-lymphocyte proliferation.

[0105] According to another embodiment of the present invention the immunosuppressive polypeptides are capable of suppressing the immune response in an animal suffering from a general skin inflammation according to the TPA model, an irritant contact dermatitis model, as described herein below. According to the present invention, the immunosuppressive polypeptides of the present invention are capable of reducing the ear thickening in mice challenged with phorbol 12-myristate 13-acetate (TPA), the ear thickening being reduced with at least 5%, such as least 10%, at least, 15%, at least 20%, such as at least 25%, at least 30%, at least 35%, such as at least 40%, at least 45%, such as at least 50%, at least 55%, such as at least 60%, at least 65%, such as at least 70%, at least 75%, such as at least 80%, at least 85% reduction of ear thickening following TPA challenge.

[0106] Hence, the present invention comprise one or more immunosuppressive peptides, such as 2, for example 3, such as 4, such as 5, for example 6, such as 7, such as 8, for example 9, such as 10, such as 11, for example 12, such as 13, such as 14, for example 15, such as 16, such as 17, for example 18, such as 19, such as 20 immunosuppressive peptides.

[0107] The present invention may comprise the same immunosuppressive polypeptide, or the compositions may comprise different immunosuppressive polypeptides. In one embodiment of the present invention, the immunosuppressive polypeptides are monomeric. In another embodiment of the present invention the immunosuppressive polypeptides are dimeric. In another embodiment of the present invention the immunosuppressive polypeptides are trimeric. In yet another embodiment of the present invention the immunosuppressive polypeptides are multimeric. Thus, according to the present invention the immunosuppressive polypeptides may be monomeric, homologous dimeric, heterologous dimeric, homologous trimeric, heterologous trimeric, homologous multimeric and/or heterologous multimeric. In a particular preferred embodiment the immunosuppressive polypeptides of the present invention are homologous dimeric.

[0108] Additionally, the present invention may comprise combinations of di-, tri-and/or multimeric immunosuppressive peptides. In one embodiment the present invention comprises homologues dimeric peptides in combination with other homologous dimeric peptides. In another embodiment the invention comprises homologous dimeric peptides in combination with heterologous dimeric peptides. The following combinations of peptides are also within the scope of this invention: homologous dimeric peptides with homologous trimeric, homologuos dimeric with heterologous trimeric, heterologous dimeric with homologous trimeric, heterologous dimeric with heterologous trimeric, homologous dimeric with homologous multimeric, heterologous dimeric with homologous multimeric, homologous dimeric with heterologous multimeric, heterologous dimeric with heterologous multimeric, homologous trimeric with homologous multimeric, homologous trimeric with heterologous multimeric, heterologous trimeric with homologous multimeric and heterologous trimeric with heterologous multimeric immusuppressive peptides.

[0109] In certain embodiments of the present invention the immunosuppressive polypeptides are homologous dimers, such as homologous dimers formed by two of the peptides SEQ ID NO: 4, and/or two of the peptides with SEQ ID NO: 119, and/or two of the peptides with SEQ ID NO: 120, and/or two of the peptides with SEQ ID NO: 121, and/or two of the peptides with SEQ ID NO: 122, and/or two of the peptides with SEQ ID NO: 123, and/or two of the peptides with SEQ ID NO: 124, and/or two of the peptides with SEQ ID NO: 125, and/or two of the peptides with SEQ ID NO: 126. In one embodiment the monomeric peptides are cross-linked into a dimer by cross-linking the peptides N-terminal to N-terminal or C-terminal to C-terminal. I a preferred embodiment the peptides are cross-linked via a disulfide bond wherein the peptides are cross-linked C-terminal to C-terminal.

[0110] In other certain embodiments of the present invention the immunosuppressive polypeptides are heterologous dimers, such as heterologus dimers formed by two peptides in the following combinations: SEQ ID NO: 4 with SEQ ID NO: 119; and/or SEQ ID NO: 4 with SEQ ID NO: 120, and/or SEQ ID NO: 4 with SEQ ID NO: 121, and/or SEQ ID NO: 4 with SEQ ID NO:122, and/or SEQ ID NO: 4 with SEQ ID NO: 123, and/or SEQ ID NO: 4 with SEQ ID NO: 124, and/or SEQ ID NO: 4 with SEQ ID NO: 125, and/or SEQ ID NO: 4 with SEQ ID NO: 126 and/or with a sequence selected from SEQ ID NO: 119 to 126 with a sequence selected from SEQ ID NO: 119 to 126.

[0111] In one embodiment the monomeric peptides are cross-linked into a dimer by cross-linking the peptides N-terminal to N-terminal or C-terminal to C-terminal. I a preferred embodiment the peptides are cross-linked via a disulfide bond wherein the peptides are cross-linked C-terminal to C-terminal.

[0112] The immunosuppressive polypeptides of the present invention may be of different length. However, it is appreciated that the active component of the immunosuppressive peptides have a maximum length of about 100 amino acids, such as about 90 amino acids, for example about 80 amino acids, such as about 70 amino acids, such as about 60 amino acids, for example about 50 amino acids, such as 40 amino acids, for example about 35 amino acids.

[0113] In particular embodiments the length of the active component of the immunosuppressive peptides is 35 amino acids, or 34, or 33, or 32, or 31, or 30, or 29, or 28, or 27, or 26, or 25, or 24, or 23, or 22, or 21, or 20, or 19, or 18, or 17, or 16, or 15, or 14, or 13, or 12, or11,or10,or9, or 8, or7, or 6, or 5, or 4, or 3 amino acids long. Thus, the immunosuppressive peptides of the present invention have lengths and amino acid sequences corresponding to any of SEQ ID NO:1 to SEQ ID NO:287 as listed herein below. A special feature of the immunosuppressive peptides of the present invention is that they may contain an extra cysteine (Cys or C) residue, either in the N-terminal or C-terminal of the polypeptide. In a particular embodiment the cysteine residue is located in the C-terminal of the peptides. The presence and function of this cysteine residue is primarily so as to crosslink two or more polypeptides together, preferable via disulfide bonds, as described herein below. However, the function of the extra cysteine may be other than that of cross-linking. Thus, the immunosuppressive peptides of the present invention may have amino acid sequences corresponding to any of SEQ ID:1 to SEQ ID:287, and wherein the immunosuppressive peptides further contain an extra cystein (Cys og C) residue at either the N-terminal or C-terminal of the peptide.

[0114] The immusuppressive peptides of the present invention may be a combination of the peptides corresponding to SEQ ID NO:1 to SEQ ID NO:287. Thus also comprise one part of one of the peptides

[0115] Moreover, the present invention also encompasses polypeptides, wherein one or more amino acid residues are modified, wherein said one or more modification(s) are preferably selected from the group consisting of in vivo or in vitro chemical derivatization, such as but not limited to acetylation or carboxylation, glycosylation, such as glycosylation resulting from exposing the polypeptide to enzymes which affect glycosylation, for example mammalian glycosylating or deglycosylating enzymes, phosphorylation, such as modification of amino acid residues which results in phosphorylated amino acid residues, for example phosphotyrosine, phosphoserine and phosphothreonine. The polypeptide according to the invention can comprise one or more amino acids independently selected from the group consisting of naturally occurring L-amino acids, naturally occurring D-amino acids as well as non-naturally occurring, synthetic amino acids. One or more amino acid residues of the polypeptide of the present invention are modified so as to preferably improve the resistance to proteolytic degradation and stability or to optimize solubility properties or to render the polypeptide more suitable as a therapeutic agent. The invention also relates to polypeptides of the invention where blocking groups are introduced in order to protect and/or stabilize the N-and/or C-termini of the polypeptide from undesirable degradation. Such blocking groups may be selected from the group comprising but not limited to branched or non-branched alkyl groups and acyl groups, such as formyl and acetyl groups, as well substituted forms thereof, such as acetamidomethyl. The invention also relates to the following: The polypeptides according to present invention, wherein the one or more blocking groups are selected from N-terminal blocking groups comprising desamino analogs of amino acids, which are either coupled to the N-terminus of the peptide or used in place of the N-terminal amino acid residue. The polypeptide according to present invention, but not limited to wherein the one or more blocking groups are selected from C-terminal blocking groups wherein the carboxyl group of the C-terminus is either incorporated or not, such as esters, ketones, and amides, as well as descarboxylated amino acid analogues. The polypeptide according to present invention, wherein the one or more blocking groups are selected from C-terminal blocking groups comprising ester or ketoneforming alkyl groups, such as lower (C1 to C6) alkyl groups, for example methyl, ethyl and propyl, and amide-forming amino groups, such as primary amines (-NH2), and mono-and dialkylamino groups, such as methylamino, ethylamino, dimethylamino, diethylamino, methylethylamino, and the like. The polypeptide according to present invention, wherein free amino group(s) at the N-terminal end and free carboxyl group(s) at the termini can be removed altogether from the polypeptide to yield desamino and descarboxylated forms thereof without significantly affecting the biological activity of the polypeptide. The increased properties may be achieved for example by chemical protection, i.e. by reacting the proteins and peptides of the present invention with protecting chemical groups, or by the incorporation of non-naturally occurring amino acids, e.g. D-amino acids, with the result of prolonging the half-life of the proteins and peptides of the present invention.

Cross-linking

[0116] The immunosuppressive polypeptides of the present invention are suitably used alone, but is preferably coupled to another material or cross-linked to itself to increase its biological or immunological activity, particularly if the polypeptide is relatively short, or to achieve certain properties on the material being coupled. In a specific aspect of this invention, any or all of the immunosuppressive polypeptides may be cross-linked to increase its activity, to facilitate its delivery in vivo, and/or to render the polypeptides resistant towards hydrolysis and/or proteolysis. The cross-linked polypeptide may be formed in situ by allowing the monomers to oxidize (e.g., for disulfide bonds) or it may be synthesized by using a specific cross-linking agent.

[0117] The cross-linking between the polypeptide chains may occur at either end of the polypeptide, or in the middle of the polypeptide, depending on which end is most appropriate. For example, if the N-terminal or the C-terminal of the polypeptides comprises cysteine residues, these are preferably cross-linked by linking it to another cysteine residue on another homologous or heterologous polypeptide of the present invention, thereby forming a disulfide bond. Preferably the immunosuppressive polypeptides of the present invention are cross-linked by disulfide bonds at the C-terminal.

[0118] Polypeptide chains may be polymerized by cross-linking agents, either directly or indirectly through multifunctional polymers. Two polypeptides may be cross linked at their C-or N-termini using a multifunctional cross-linking agent. The agent is used to cross-link the terminal amino-or carboxyl groups. Generally, both terminal carboxyl groups or both terminal amino groups are crosslinked to one another, although by selection of the appropriate crosslinking agent the alpha amino group of one polypeptide is crosslinked to the terminal carboxyl group of the other polypeptide. Preferably, the polypeptides are substituted at their C-termini with cysteine. Under conditions well known in the art a disulfide bond can be formed between the terminal cysteines, thereby cross-linking the polypeptide chains.

[0119] Additional cross-linking sites on the polypeptides, include epsilon amino groups found on lysine residues, as well as amino, imino, carboxyl, sulfhydryl and hydroxyl groups located on the side chains of internal residues of the peptides. Cross-linking through externally added cross-linking agents is obtained, e.g., using any of a number of reagents familiar to those skilled in the art, for example, via carbodiimide treatment of the polypeptides. Other non-limiting examples of suitable multifunctional cross-linking agents include 1,1-bis(diazoacetyl)-2-phenylethane; glutaraldehyde; Nhydroxysuccinimide esters such as esters with 4-azidosalicylic acid; homobifunctional imidoesters including disuccinimidyl esters such as 3,3′-dithiobis (succinimidylpropionate) and dimethyl adipimidate dihydrochloride, bifunctional maleimides such as bis-N-maleimidol,8-octane; disuccinimidyl suberate, and bis(sulfosuccinimidyl) suberate.

[0120] Heterobifunctional cross-linking reagents include those with an N-hydroxysuccinimide moiety at one end and a maleimido group on the other end; succinimidyl 4-(Nmaleimidomethyl)cyclohexane-1-carboxylate (SMCC), sulfo-SMCC, mmaleimidobenzoyl-N-hydroxysuccinimide ester (MBS); sulfo-MBS; suceinimidyl 4-(pmaleimidophenyl)butyrate (SMPB); sulfo-SMPB; N-succinimidyl(4-iodoacetyl)aminobenzoate (SlAB); sulfo-SIAB; 1-ethyl-3-(3dimethylaminopropyl)carbodiimide hydrochloride (EDC); and Nhydroxysulfosuccinimide. Cross-linking agents such as methyl-3-[(p-azido-phenyl)dithio) propioimidate yield photoactivatable intermediates which are capable of forming cross-links in the presence of light. If necessary, sensitive residues such as the side chains of the diargininyl group are protected during cross-linking and the protecting groups removed thereafter.

[0121] Polymers capable of multiple cross-linking serve as indirect cross-linking agents. For example, cyanogen bromide activated carbohydrates may be used for cross-linking the peptides herein. Cross-linking to amino groups of the peptides is accomplished by known chemistries based upon eyanuric chloride, carbonyl diimidazole, aldehyde reactive groups (PEG alkoxide plus diethyl acetal of bromoacetaldehyde; PEG plus DMSO and acetic anhydride, or PEG chloride plus the phenoxide of 4-hydroxybenzaldehyde). Also useful are succinimidyl active esters, activated dithiocarbonate PEG, and 2,4,5-trichlorophenylchloroformate-or pnitrophenylchloroformate-activated PEG. Carboxyl groups are derivatized by coupling PEG-amine using carbodiimide.

Administration Forms, Formulations and Dosage Regimes

[0122] Pharmaceutical compositions containing a composition of the present invention may be prepared by conventional techniques, e.g. as described in Remington: The Science and Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton, Pa. The compositions may appear in conventional forms, for example suspensions or topical applications such as a solution, gel, cream, lotion, shake lotion, ointment, foam, shampoo, mask or similar forms. But also patches, gazes and bandages and the like may be used for topical application of the composition of the present invention.

[0123] Whilst it is possible for the compositions or salts of the present invention to be administered as the raw chemical, it is preferred to present them in the form of a pharmaceutical formulation. Accordingly, the present invention further provides a pharmaceutical formulation, for medicinal application, which comprises a composition of the present invention or a pharmaceutically acceptable salt thereof, as herein defined, and a pharmaceutically acceptable carrier therefore.

[0124] The pharmaceutical compositions and dosage forms may comprise the compositions of the invention or its pharmaceutically acceptable salt or a crystal form thereof as the active component. The pharmaceutically acceptable carriers can be either solid, semisolid or liquid. Emulsions may be prepared in solutions in aqueous propylene glycol solutions or may contain emulsifying agents such as lecithin, sorbitan monooleate, or acacia. Aqueous solutions can be prepared by suspending or mixing the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents. Aqueous suspensions can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well known suspending agents. Solid form preparations include suspensions and emulsions, and may contain, in addition to the active component, colorants, stabilizers, buffers, artificial and natural dispersants, thickeners, and the like.

[0125] The compositions of the present invention may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, for example solutions in aqueous polyethylene glycol. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and may contain formulatory agents such as preserving, wetting, emulsifying or suspending, stabilizing or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilisation from solution for constitution before use with a suitable vehicle, e.g., sterile, pyrogen-free water.

[0126] Oils useful in formulations include petroleum, animal, vegetable, or synthetic oils. Specific examples of oils useful in such formulations include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral. Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0127] Suitable soaps for use in formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents such as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides; (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl.beta.-aminopropionates, and 2-alkyl-imidazoline quaternary ammonium salts, and (e) mixtures thereof.

[0128] The formulations typically will contain from about 0.5 to about 25% by weight of the active ingredient in solution. Preservatives and buffers may be used. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations will typically range from about 5 to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The parenteral formulations can be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, immediately prior to use.

Pharmaceutically Acceptable Salts

[0129] Pharmaceutically acceptable salts of the instant compositions, where they can be prepared, are also intended to be covered by this invention. These salts will be ones which are acceptable in their application to a pharmaceutical use. By that it is meant that the salt will retain the biological activity of the parent composition and the salt will not have untoward or deleterious effects in its application and use in treating diseases.

[0130] Pharmaceutically acceptable salts are prepared in a standard manner. If the parent composition is a base it is treated with an excess of an organic or inorganic acid in a suitable solvent. If the parent composition is an acid, it is treated with an inorganic or organic base in a suitable solvent.

[0131] The compositions of the invention may be administered in the form of an alkali metal or earth alkali metal salt thereof, concurrently, simultaneously, or together with a pharmaceutically acceptable carrier or diluent, especially and preferably in the form of a pharmaceutical composition thereof, whether by oral, rectal, or parenteral (including subcutaneous) route, in an effective amount.

[0132] Examples of pharmaceutically acceptable acid addition salts for use in the present inventive pharmaceutical composition include those derived from mineral acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, and organic acids, such as tartaric, acetic, citric, malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic, p-toluenesulphonic acids, and arylsulphonic, for example.

Uses of Compositions Containing Said Immunosuppressive Peptides

[0133] The present invention comprises in one embodiment a pharmaceutical composition and/or compositions for the treatment and/or prevention and/or amelioration of inflammatory disorders. Below is a non-limiting list of the inflammatory disorders that the compositions of the present invention can be used to treat, prevent or ameliorate. The compositions of the present invention may be directed towards the treatment, prevention or amelioration of other inflammatory disorders than the ones listed herein below. The list below may thus be regarded as the inflammatory disorders that in preferred embodiments are target conditions for the compositions of the present invention.

[0134] We anticipate that the immunosuppressive peptides disclosed in this application will be advantageous for treatment of many other types of inflammatory disorders where a reduction of anti-inflammatory responses in the patient is desirable. This is especially valid for diseases/applications where a reduction in the level of cytokines like TNF-α, IL-17, IL-6. Especially for diseases/applications like Arthritis, Asthma, Autoimmune diseases, Sepsis, Inflammatory bowel disease, Coating of biomaterials and nanoparticles where a reduction of one or several of these cytokines has been reported as desirable.

[0135] Below a number of such inflammatory disorders where a decreased immunogenic response is required is described in more detail. The description of relevant diseases should only be considered as examples as many more diseases could be treated these immunosuppressive peptides. Also included is the usage of these immunosuppressive peptides for coating of nanoparticles and biomaterials as a decreased immunogenic response is also desired in these cases to prolong the half-life of these materials, increase biocompatibility or decrease foreign body reactions.

Sepsis

[0136] Sepsis is a potentially deadly medical condition characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome or SIRS) that is triggered by an infection. The body may develop this inflammatory response by the immune system to microbes in the blood, urine, lungs, skin, or other tissues. A lay term for sepsis is blood poisoning, also used to describe septicaemia. Severe sepsis is the systemic inflammatory response, infection and the presence of organ dysfunction.

[0137] Severe sepsis is usually treated in the intensive care unit with intravenous fluids and antibiotics. If fluid replacement isn’t sufficient to maintain blood pressure, specific vasopressor medications can be used. Mechanical ventilation and dialysis may be needed to support the function of the lungs and kidneys, respectively. To guide therapy, a central venous catheter and an arterial catheter may be placed; measurement of other hemodynamic variables (such as cardiac output, mixed venous oxygen saturation, or stroke volume variation) may also be used. Sepsis patients require preventive measures for deep vein thrombosis, stress ulcers and pressure ulcers, unless other conditions prevent this. Some patients might benefit from tight control of blood sugar levels with insulin (targeting stress hyperglycemia). The use of corticosteroids (low dose or otherwise) is controversial. Activated drotrecogin alfa (recombinant protein C) has not been found to be helpful, and has recently been withdrawn from sale.

[0138] In addition to symptoms related to the provoking infection, sepsis is characterized by presence of acute inflammation present throughout the entire body, and is, therefore, frequently associated with fever and elevated white blood cell count (leukocytosis) or low white blood cell count (leukopenia) and lower-than-average temperature, and vomiting. The modern concept of sepsis is that the host’s immune response to the infection causes most of the symptoms of sepsis, resulting in hemodynamic consequences and damage to organs. This host response has been termed systemic inflammatory response syndrome (SIRS) and is characterized by an elevated heart rate (above 90 beats per minute), high respiratory rate (above 20 breaths per minute or a partial pressure of carbon dioxide in the blood of less than 32), abnormal white blood cell count (above 12,000, lower than 4,000, or greater than 10% band forms) and elevated or lowered body temperature, i.e. under 36° C. (96.8° F.) or over 38° C. (100.4° F.). Sepsis is differentiated from SIRS by the presence of a known or suspected pathogen. For example SIRS and a positive blood culture for a pathogen indicates the presence of sepsis. However, in many cases of sepsis no specific pathogen is identified.

[0139] This immunological response causes widespread activation of acute-phase proteins, affecting the complement system and the coagulation pathways, which then cause damage to the vasculature as well as to the organs. Various neuroendocrine counter-regulatory systems are then activated as well, often compounding the problem. Even with immediate and aggressive treatment, this may progress to multiple organ dysfunction syndrome and eventually death.

[0140] The term “amino acid” and “amino acid sequence” refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where “amino acid sequence” is recited to refer to a sequence of a naturally occurring protein molecule, “amino acid sequence” and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule. Thus, the term “amino acid” comprises any synthetic or naturally occurring amino carboxylic acid, including any amino acid occurring in peptides and polypeptides including proteins and enzymes synthesized in vivo thus including modifications of the amino acids. The term amino acid is herein used synonymously with the term “amino acid residue” which is meant to encompass amino acids as stated which have been reacted with at least one other species, such as 2, for example 3, such as more than 3 other species. The generic term amino acid comprises both natural and non-natural amino acids any of which may be in the “D” or “L” isomeric form.

TABLE-US-00001 One-letter symbol Three-letter symbol Amino acid (trivial name) A Ala alanine B Asx aspartic acid or asparagine C Cys cysteine D Asp aspartic acid E Glu glutamic acid F Phe phenylalanine G Gly glycine H His histidine I Ile isoleucine K Lys lysine L Leu leucine M Met methionine N Asn asparagine P Pro proline Q Gln glutamine R Arg arginine S Ser serine T Thr threonine U* Sec selenocysteine V Val valine W Trp tryptophan X Xaa unknown or other amino acid, i.e. X can be any of the conventional amino acids. Y Tyr Tyrosine Z Glx glutamic acid or glutamine (or substances such as 4-carboxyglutamic acid and 5-oxoproline that yield glutamic acid on acid hydrolysis of peptides)

[0141] The term “polypeptide” refers to a peptide having at least two, and preferably more than two amino acids. “Monomeric polypeptide” refers to a polypeptide that is a monomer as opposed to a dimer, trimer or polymer in the sense that the monomeric polypeptide is not crosslinked or otherwise bonded to another polypeptide chain. The term “dimer” thus refers to a moiety wherein two monomeric polypeptides are crosslinked to each other. In the same way, a trimeric polypetide refers to a moiety wherein three monomeric polypeptides are crosslinked to each other, while the term “polymer” or “multimer” refers to a moiety wherein at least two polypeptides, and preferably more than three polypeptides are crosslinked to each other.

[0142] The expression “cross-linker” or “cross-linking moiety” refers to a linking moiety conferred by an external cross-linking agent used to crosslink one polypeptide with one or more polypeptides as described further in detail herein below.

[0143] The term “homology” refers to sequence similarity or, interchangeably, sequence identity, between two or more polynucleotide sequences or two or more polypeptide sequences.

[0144] The phrases “percent identity” and “% identity,” as applied to polypeptide sequences, refer to the percentage of residue matches between at least two polypeptide sequences aligned using a standardized algorithm. Methods of polypeptide sequence alignment are well-known. Some alignment methods take into account conservative amino acid substitutions. Such conservative substitutions, explained in more detail above, generally preserve the charge and hydrophobicity at the site of substitution, thus preserving the structure (and therefore function) of the polypeptide.

[0145] “Percent identity” may be measured over the length of an entire defined polypeptide sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 6, at least 8, at least 10, at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

[0146] The term “carrier” refers to a compound that is conjugated to the polypeptide(s) either to increase the number of polypeptides, for increasing activity or immunosuppressive effect of the polypeptide(s), to confer stability to the molecules, to increase the biological activity of the peptides, or to increase its serum half-life. The “carrier” may be a protein carrier or a non-protein carrier. Non-limiting examples of non-protein carriers include liposomes, micelles, polymeric nanoparticles and diaminoethane. The liposome may comprise glycosaminoglycan hyaluronan (HA) and/or PEG. In one embodiment, the carrier is an immunoliposome. Other carriers include protamines,or polysaccharides e.g. aminodextran or chitosan. Non-limiting examples of protein carriers include, keyhole limpet hemocyanin, serum proteins such as transferrin, bovine serum albumin, human serum albumin, whale myoglobin, ovalbumn, immunoglobulins, lysozyme, carbonic anhydrase, or hormones, such as insulin. In other embodiments of the present invention, the carrier may be a pharmaceutical acceptable carrier as described herein below. The immunosuppressive peptides of the present invention may be coupled to the carrier by means of cross-linking as further described herein below.

[0147] The terms “protein modification”, “protein stability” and “peptide stability” is used to describe the state of the immunosuppressive proteins and peptides, in particular the state wherein said proteins and/or peptides are more resistant to degradation and/or have increased properties towards hydrolysis and/or proteolysis. In particular, proteolytic stability refers to the resistance toward the action of proteolytic enzymes, also known as proteases, i.e. enzymes that catalyzes the hydrolysis of the amide/peptide-bond of the protein or peptide. Moreover, the present invention also encompasses polypeptides, wherein one or more amino acid residues are modified, wherein said one or more modification(s) are preferably selected from the group consisting of in vivo or in vitro chemical derivatization, such as but not limited to acetylation or carboxylation, glycosylation, such as glycosylation resulting from exposing the polypeptide to enzymes which affect glycosylation, for example mammalian glycosylating or deglycosylating enzymes, phosphorylation, such as modification of amino acid residues which results in phosphorylated amino acid residues, for example phosphotyrosine, phosphoserine and phosphothreonine. The polypeptide according to the invention can comprise one or more amino acids independently selected from the group consisting of naturally occurring L-amino acids, naturally occurring D-amino acids as well as non-naturally occurring, synthetic amino acids. One or more amino acid residues of the polypeptide of the present invention are modified so as to preferably improve the resistance to proteolytic degradation and stability or to optimize solubility properties or to render the polypeptide more suitable as a therapeutic agent. The invention also relates to polypeptides of the invention where blocking groups are introduced in order to protect and/or stabilize the N-and/or C-termini of the polypeptide from undesirable degradation. Such blocking groups may be selected from the group comprising but not limited to branched or non-branched alkyl groups and acyl groups, such as formyl and acetyl groups, as well substituted forms thereof, such as acetamidomethyl. The invention also relates to the following: The polypeptides according to present invention, wherein the one or more blocking groups are selected from N-terminal blocking groups comprising desamino analogs of amino acids, which are either coupled to the N-terminus of the peptide or used in place of the N-terminal amino acid residue. The polypeptide according to present invention, but not limited to wherein the one or more blocking groups are selected from C-terminal blocking groups wherein the carboxyl group of the C-terminus is either incorporated or not, such as esters, ketones, and amides, as well as descarboxylated amino acid analogues. The polypeptide according to present invention, wherein the one or more blocking groups are selected from C-terminal blocking groups comprising ester or ketoneforming alkyl groups, such as lower (C1 to C6) alkyl groups, for example methyl, ethyl and propyl, and amide-forming amino groups, such as primary amines (-NH2), and mono-and dialkylamino groups, such as methylamino, ethylamino, dimethylamino, diethylamino, methylethylamino, and the like. The polypeptide according to present invention, wherein free amino group(s) at the N-terminal end and free carboxyl group(s) at the termini can be removed altogether from the polypeptide to yield desamino and descarboxylated forms thereof without significantly affecting the biological activity of the polypeptide. The increased properties may be achieved for example by chemical protection, i.e. by reacting the proteins and peptides of the present invention with protecting chemical groups, or by the incorporation of non-naturally occurring amino acids, e.g. D-amino acids, with the result of prolonging the half-life of the proteins and peptides of the present invention.

[0148] The term “penetration promoting” or “penetration enhancing” as used herein refers to compounds that facilitate the delivery of the immunosuppressive peptides of the present invention to the target site of action. In particular the term refers to the transcutaneous delivery of the immunosuppressive peptides. Simple topical application of the present invention may not always yield an adequate result, as the outermost layer of the skin provides an outstanding barrier against the external environment. While single penetration enhancers can aid topical delivery, combinations of several penetration enhancers may most effective. The amount of penetration enhancer which may be used in the invention varies from about 1 to 100 percent although adequate enhancement of penetration is generally found to occur in the range of about 1 to about 10 percent by weight of the formulation to be delivered. Non-limiting examples of penetration enahancers are entities that falls within liposomes, transferosomes niosomes and ethosomes, but may also be any of the many hundred known chemical prentration enhancers, of which sulfoxides, azones, pyrrolidones, fatty acids, terpenes and terpenoids, oxazolidinones and urea are non-limiting examples.

[0149] The term “immuno-modulation” as used herein refers to the process of where an immune response is either suppressed, partly or completely, or triggered or induced or enhanced. Likewise, the term “growth-modulation” as used herein refers to the process of were the cell proliferation is either suppressed, partly or completely, or where cell proliferation is induced or enhanced or promoted.

[0150] The term “substance” as used anywhere herein comprises any form of substance suitable for comprising the immunosuppressive polypeptides of the present invention.

[0151] Non-limiting examples of such substances are creams, lotions, shake lostions, ointments, gels, balms, salves, oils, foams, shampoos, sprays, aerosoloes as well as transdermal patches and bandages.

[0152] The term “treatment”, as used anywhere herein comprises any type of therapy, which aims at terminating, preventing, ameliorating and/or reducing the susceptibility to a clinical condition as described herein. In a preferred embodiment, the term treatment relates to prophylactic treatment, i.e. a therapy to reduce the susceptibility of a clinical condition, a disorder or condition as defined herein.

[0153] Thus, “treatment,” “treating,” and the like, as used herein, refer to obtaining a desired pharmacologic and/or physiologic effect, covering any treatment of a pathological condition or disorder in a mammal, including a human. The effect may be prophylactic in terms of completely or partially preventing a disorder or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disorder and/or adverse affect attributable to the disorder. That is, “treatment” includes (1) preventing the disorder from occurring or recurring in a subject, (2) inhibiting the disorder, such as arresting its development, (3) stopping or terminating the disorder or at least symptoms associated therewith, so that the host no longer suffers from the disorder or its symptoms, such as causing regression of the disorder or its symptoms, for example, by restoring or repairing a lost, missing or defective function, or stimulating an inefficient process, or (4) relieving, alleviating, or ameliorating the disorder, or symptoms associated therewith, where ameliorating is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, such as inflammation, pain, and/or immune deficiency.

[0154] The term “animal” as used herein may be defined to include humans, domestic or agricultural (cats, dogs, cows, sheep, horses, pigs, etc.) or test species such as mouse, rats, rabbits and the like. Thus the anamals may also be of bovine, equine, porcine, human, ovine, caprine or cervidae origin.

[0155] According to an embodiment, the present invention concerns an immune suppressive domain for use as a medicament.

[0156] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is the fusion peptide of an envelope protein.

[0157] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is the fusion peptide of a virus.

[0158] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is the fusion peptide of an enveloped RNA virus.

[0159] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is from a virus.

[0160] The inventors have inter alia identified three new groups of enveloped RNA viruses with immunosuppressive domains in their fusion protein: [0161] 1: The inventors have identified immunosuppressive domains among enveloped RNA viruses with type II fusion mechanism. Hitherto, immunosuppressive domains have not been described for any enveloped RNA viruses with a type II fusion mechanism. Immunosuppressive domains have been identified by the inventors at two positions in two different groups of viruses: [0162] i. Co-localizing with the fusion peptide exemplified by the identification of an common immunosuppressive domain in the fusion peptide of Flavirius (Dengue virus, west Nile virus etc), and [0163] ii. In the hydrophobic alpha helix N-terminal of the transmembrane domain in the fusion protein exemplified by the finding of an immunosuppressive domain in said helixes of all flaviridae e.g. Hepatitis C virus, Dengue, west nile etc. [0164] 2: The inventors have identified immunosuppressive domains in the fusion protein among enveloped RNA viruses with type I fusion mechanism (excluding lentivirus, retrovirus and filovirus). [0165] This position co-localizes with the fusion peptide of said fusion protein as demonstrated by the identification of a common immunosuppressive domain in the fusion peptide of all Influenza A and B types. [0166] 3: The inventors have identified potential immunosuppressive domains located at various positions of type I enveloped RNA viruses (excluding lentivirus, retrovirus and filovirus) as well as in enveloped RNA viruses featuring a fusion protein with neither a type I nor a type II fusion structure.

[0167] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is from an influenza virus.

[0168] According to an embodiment, the invention concerns the immune suppressive domain, wherein said domain is derived from an enveloped RNA virus.

[0169] The expression “derived from a virus” means that the domain is substantially identical to the immune suppressive domain of the virus, optionally with mutations, insertions or deletions.

[0170] According to an embodiment, the invention concerns the immune suppressive domain, subject to the proviso that said immune suppressive domain is different from immunosuppressive domains obeying the conditions of: [0171] i) being from a virus selected among the group consisting of gammaretrovirus, HIV and filovirus; [0172] ii) being located in the linker between the two heptad repeat structures just N-terminal of the transmembrane domain in the fusion protein; and [0173] iii) including at least some of the first 22 amino acids located N-terminal to the first cysteine residue of the two well conserved cystein residues, located between 4 and 6 amino acid residues from one another and further located just N-terminal of the transmembrane spanning domain of the fusion protein.

[0174] The immunosuppressive domains of lentivirus, retroviruses and filoviruses show large structural similarity. Furthermore the immunosuppressive domain of these viruses are all located at the same position in the structure of the fusion protein, more precisely in the linker between the two heptad repeat structures just N-terminal of the transmembrane domain in the fusion protein. These heptad repeat regions constitute two alpha helices that play a critical role in the active mechanism of membrane fusion by these proteins. The immune suppressive domains can be located in relation to two well conserved cystein residues that are found in these structures. These cystein residues are between 4 and 6 amino acid residues from one another and in many cases are believed to form disulfide bridges that stabilize the fusion proteins. The immune suppressive domains in all three cases include at least some of the first 22 amino acids that are located N-terminal to the first cysteine residue.

[0175] According to an embodiment, the invention concerns the immune suppressive domain, subject to the proviso, that said immune suppressive domain is different from immunosuppressive domains obeying the conditions of: [0176] i) being from a virus selected among the group consisting of gammaretrovirus, HIV and filovirus; and [0177] ii) being located in the linker between the two heptad repeat structures just N-terminal of the transmembrane domain in the fusion protein.

[0178] The in this context relevant immunosuppressive domains are all located at a very well-defined structure within their fusion proteins, at the bend in the heptad repeat just N-terminale of the transmembrane structure in the fusion protein.

[0179] According to an embodiment, the invention concerns the immune suppressive domain, wherein said immune suppressive domain is different from immunosuppressive domains from a virus selected among the group consisting of gammaretrovirus, HIV and filovirus.

[0180] According to an embodiment, the invention concerns the immune suppressive domain, wherein the domain is selected among the sequences of Table 1 or sequences seqid 1 to seqid 287.

[0181] According to an embodiment, the invention concerns the immune suppressive domain, wherein the domain is obtainable from the sequences of Table 1 or the sequences seqid 1 to seqid 287, by at least one mutation, deletion or insertion.

[0182] According to an embodiment, the invention concerns the immune suppressive domain, wherein the total number of mutations, deletions or insertions is selected among 1, 2, 3 and 4.

[0183] The term “mutation” is used with a number about this number of point mutation(s), i.e. 3 mutations mean 3 point mutations. The term “deletion” is used with a number about the deletion of this number of amino acid(s), i.e. 2 deletions means the deletion of 2 amino acids. The term “insertion” is used with a number about insertion of this number of amino acid(s), i.e. 1 insertion means the insertion of 1 amino acid.

[0184] According to an embodiment, the invention concerns the immune suppressive domain, wherein the total number of mutations, deletions or insertions is more than 4.

[0185] According to an embodiment, the invention concerns the immune suppressive domain, whereby the obtained immune suppressive domain have abrogated immunosuppressive properties for use in a vaccine against Porcine Reproductive and Respiratory Syndrom (PRRS).

[0186] According to an embodiment, the invention concerns the immune suppressive domain for use in surgery, prophylaxis, therapy, or a diagnostic method.

[0187] According to an embodiment, the invention concerns the immune suppressive domain, wherein the domain is selected among the group consisting of segid 4 and seqid 119 to seqid 126.

[0188] According to an embodiment, the invention concerns the immune suppressive domain, wherein the domain is homologous to seqid 4.

[0189] According to an embodiment, the invention concerns the immune suppressive domain, which is a monomeric peptide.

[0190] According to an embodiment, the invention concerns the immune suppressive domain, cross-linked to at least one additional immunosuppressive peptide.

[0191] According to an embodiment, the invention concerns the immune suppressive domain, connected to at least one additional immunosuppressive peptide to form a dimer.

[0192] According to an embodiment, the invention concerns the immune suppressive domain, wherein said dimer is homologous and comprises at least two immunosuppressive peptides with SEQ ID NO. 4, which are cross-linked by a disulfide bond, N-terminal to N-terminal or C-terminal to C-terminal.

[0193] According to an embodiment, the invention concerns the immune suppressive domain, wherein said dimer is homologous and comprises at least two immunosuppressive peptides selected from SEQ ID NO. 119 to seqid 126, which are cross-linked by a disulfide bond, N-terminal to N-terminal or C-terminal to C-terminal.

[0194] According to an embodiment, the invention concerns the immune suppressive domain, connected to at least one additional immunosuppressive peptide to form a heterologous dimer.

[0195] According to an embodiment, the invention concerns the immune suppressive domain, connected to at least two additional immunosuppressive peptides to form a multimer.

[0196] According to an embodiment, the invention concerns the immune suppressive domain, wherein said immunosuppressive peptides comprises one or more modifications.

[0197] According to an embodiment, the invention concerns the immune suppressive domain, wherein said modifications are selected from the group consisting of chemical derivatizations, L-amino acid substitutions, D-amino acid substitutions, synthetic amino acid substitutions, deaminations and decarboxylations.

[0198] According to an embodiment, the invention concerns the immune suppressive domain, wherein the peptides or proteins have increased resistance against proteolysis compared to peptides or proteins not comprising said at least one modification.

[0199] According to an embodiment, the invention concerns the immune suppressive domain or an immune suppressive peptide according to the invention, for use in diagnostics and/or treatment and/or prevention and/or amelioration of disease.

[0200] According to an embodiment, the invention concerns the immune suppressive domain, wherein the subject is a human or an animal.

[0201] According to an embodiment, the invention concerns the immune suppressive domain, for use on an organ.

[0202] It is envisaged that an ISD may be used for treating an organ, e.g. before transplantation.

[0203] According to an embodiment, the invention concerns the immune suppressive domain for immune suppression.

[0204] According to an embodiment, the invention concerns the immune suppressive domain for the preparation or treatment of transplantation patients.

[0205] According to an embodiment, the invention concerns the immune suppressive domain for a use comprising treatment and/or prevention and/or amelioration of an autoimmune or inflammatory disease.

[0206] According to an embodiment, the invention concerns the immune suppressive domain for a use comprising prophylaxis or treatment of a condition selected among Acute disseminated encephalomyelitis (ADEM), Addison’s disease, Agammaglobulinemia, Alopecia areata, Amyotrophic Lateral Sclerosis, Ankylosing Spondylitis, Antiphospholipid syndrome, Antisynthetase syndrome, Atopic allergy, Atopic dermatitis, Autoimmune aplastic anemia, Autoimmune cardiomyopathy, Autoimmune enteropathy, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune lymphoproliferative syndrome, Autoimmune peripheral neuropathy, Autoimmune pancreatitis, Autoimmune polyendocrine syndrome, Autoimmune progesterone dermatitis, Autoimmune thrombocytopenic purpura, Autoimmune urticaria, Autoimmune uveitis, Balo disease/Balo concentric sclerosis, Behçet’s disease, Berger’s disease, Bickerstaff’s encephalitis, Blau syndrome, Bullous pemphigoid, Cancer, Castleman’s disease, Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy, Chronic recurrent multifocal osteomyelitis, Chronic obstructive pulmonary disease, Churg-Strauss syndrome, Cicatricial pemphigoid, Cogan syndrome, Cold agglutinin disease, Complement component 2 deficiency, Contact dermatitis, Cranial arteritis, CREST syndrome, Crohn’s disease, Cushing’s Syndrome, Cutaneous leukocytoclastic angiitis, Dego’s disease, Dercum’s disease, Dermatitis herpetiformis, Dermatomyositis, Diabetes mellitus type 1, Diffuse cutaneous systemic sclerosis, Dressler’s syndrome, Drug-induced lupus, Discoid lupus erythematosus, Eczema, Endometriosis, Enthesitis-related arthritis, Eosinophilic fasciitis, Eosinophilic gastroenteritis, Epidermolysis bullosa acquisita, Erythema nodosum, Erythroblastosis fetalis, Essential mixed cryoglobulinemia, Evan’s syndrome, Fibrodysplasia ossificans progressiva, Fibrosing alveolitis, Gastritis, Gastrointestinal pemphigoid, Glomerulonephritis, Goodpasture’s syndrome, Graves’ disease, Guillain-Barre syndrome (GBS), Hashimoto’s encephalopathy, Hashimoto’s thyroiditis, Henoch-Schonlein purpura, Herpes gestationis, Hidradenitis suppurativa, Hughes-Stovin syndrome, Hypogammaglobulinemia, Idiopathic inflammatory demyelinating diseases, Idiopathic pulmonary fibrosis, Idiopathic thrombocytopenic purpura, IgA nephropathy, Inclusion body myositis, Chronic inflammatory demyelinating polyneuropathy, Interstitial cystitis, Juvenile idiopathic arthritis, Kawasaki’s disease, Lambert-Eaton myasthenic syndrome, Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Linear IgA disease (LAD), Lou Gehrig’s disease, Lupoid hepatitis, Lupus erythematosus, Majeed syndrome, Ménière’s disease, Microscopic polyangiitis, Miller-Fisher syndrome, Mixed connective tissue disease, Morphea, Mucha-Habermann disease, Multiple sclerosis, Myasthenia gravis, Myositis, Narcolepsy, Neuromyelitis optica, Neuromyotonia, Occular cicatricial pemphigoid, Opsoclonus myoclonus syndrome, Ord’s thyroiditis, Palindromic rheumatism, PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcus), Paraneoplastic cerebellar degeneration, Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome, Parsonage-Turner syndrome, Pars planitis, Pemphigus vulgaris, Pernicious anaemia, Perivenous encephalomyelitis, POEMS syndrome, Polyarteritis nodosa, Polymyalgia rheumatica, Polymyositis, Primary biliary cirrhosis, Primary sclerosing cholangitis, Progressive inflammatory neuropathy, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum, Pure red cell aplasia, Rasmussen’s encephalitis, Raynaud phenomenon, Relapsing polychondritis, Reiter’s syndrome, Restless leg syndrome, Retroperitoneal fibrosis, Rheumatoid arthritis, Rheumatic fever, Sarcoidosis, Schizophrenia, Schmidt syndrome, Schnitzler syndrome, Scleritis, Scleroderma, Serum Sickness, Sjogren’s syndrome, Spondyloarthropathy, Still’s disease, Stiff person syndrome, Subacute bacterial endocarditis (SBE), Susac’s syndrome, Sweet’s syndrome, Sydenham chorea, Sympathetic ophthalmia, Systemic lupus erythematosis, Takayasu’s arteritis, Temporal arteritis, Thrombocytopenia, Tolosa-Hunt syndrome, Transverse myelitis, Ulcerative colitis, Undifferentiated connective tissue disease, Undifferentiated spondyloarthropathy, Urticarial vasculitis, Vasculitis, Vitiligo, and Wegener’s granulomatosis.

[0207] According to an embodiment, the invention concerns the immune suppressive domain for the treatment or prevention of acute or chronic inflammation.

[0208] According to an embodiment, the invention concerns the immune suppressive domain for the treatment or prevention of a disorder associated with inflammation.

[0209] According to an embodiment, the invention concerns the immune suppressive domain for the treatment or prevention of a disorder selected among Acne vulgaris, Allergy, Allergic rhinitis, Asthma, Atherosclerosis, Autoimmune disease, Celiac disease, Chronic prostatitis, Glomerulonephritis, Hypersensitivities, Inflammatory bowel diseases, Pelvic inflammatory disease, Reperfusion injury, Rheumatoid arthritis, Sarcoidosis, Transplant rejection, Vasculitis, interstitial cystitis, Cancer, Depression, Myopathies, and Leukocyte defects. These conditions are examples of diseases or conditions associated with inflammation.

[0210] According to an embodiment, the invention concerns the immune suppressive domain for a use comprising prophylaxis or treatment of sepsis.

[0211] According to an embodiment, the invention concerns the immune suppressive domain for a use comprising prophylaxis or treatment of asthma.

[0212] According to an embodiment, the invention concerns the immune suppressive domain for a use comprising prophylaxis or treatment of allergy.

[0213] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for immune suppression.

[0214] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for the preparation or treatment of transplantation patients.

[0215] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of an autoimmune or inflammatory disease.

[0216] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of a condition selected among Acute disseminated encephalomyelitis (ADEM), Addison’s disease, Agammaglobulinemia, Alopecia areata, Amyotrophic Lateral Sclerosis, Ankylosing Spondylitis, Antiphospholipid syndrome, Antisynthetase syndrome, Atopic allergy, Atopic dermatitis, Autoimmune aplastic anemia, Autoimmune cardiomyopathy, Autoimmune enteropathy, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune lymphoproliferative syndrome, Autoimmune peripheral neuropathy, Autoimmune pancreatitis, Autoimmune polyendocrine syndrome, Autoimmune progesterone dermatitis, Autoimmune thrombocytopenic purpura, Autoimmune urticaria, Autoimmune uveitis, Balo disease/Balo concentric sclerosis, Behçet’s disease, Berger’s disease, Bickerstaff’s encephalitis, Blau syndrome, Bullous pemphigoid, Cancer, Castleman’s disease, Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy, Chronic recurrent multifocal osteomyelitis, Chronic obstructive pulmonary disease, Churg-Strauss syndrome, Cicatricial pemphigoid, Cogan syndrome, Cold agglutinin disease, Complement component 2 deficiency, Contact dermatitis, Cranial arteritis, CREST syndrome, Crohn’s disease, Cushing’s Syndrome, Cutaneous leukocytoclastic angiitis, Dego’s disease, Dercum’s disease, Dermatitis herpetiformis, Dermatomyositis, Diabetes mellitus type 1, Diffuse cutaneous systemic sclerosis, Dressler’s syndrome, Drug-induced lupus, Discoid lupus erythematosus, Eczema, Endometriosis, Enthesitis-related arthritis, Eosinophilic fasciitis, Eosinophilic gastroenteritis, Epidermolysis bullosa acquisita, Erythema nodosum, Erythroblastosis fetalis, Essential mixed cryoglobulinemia, Evan’s syndrome, Fibrodysplasia ossificans progressiva, Fibrosing alveolitis, Gastritis, Gastrointestinal pemphigoid, Glomerulonephritis, Goodpasture’s syndrome, Graves’ disease, Guillain-Barre syndrome (GBS), Hashimoto’s encephalopathy, Hashimoto’s thyroiditis, Henoch-Schonlein purpura, Herpes gestationis, Hidradenitis suppurativa, Hughes-Stovin syndrome, Hypogammaglobulinemia, Idiopathic inflammatory demyelinating diseases, Idiopathic pulmonary fibrosis, Idiopathic thrombocytopenic purpura, IgA nephropathy, Inclusion body myositis, Chronic inflammatory demyelinating polyneuropathy, Interstitial cystitis, Juvenile idiopathic arthritis, Kawasaki’s disease, Lambert-Eaton myasthenic syndrome, Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Linear IgA disease (LAD), Lou Gehrig’s disease, Lupoid hepatitis, Lupus erythematosus, Majeed syndrome, Ménière’s disease, Microscopic polyangiitis, Miller-Fisher syndrome, Mixed connective tissue disease, Morphea, Mucha-Habermann disease, Multiple sclerosis, Myasthenia gravis, Myositis, Narcolepsy, Neuromyelitis optica, Neuromyotonia, Occular cicatricial pemphigoid, Opsoclonus myoclonus syndrome, Ord’s thyroiditis, Palindromic rheumatism, PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcus), Paraneoplastic cerebellar degeneration, Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome, Parsonage-Turner syndrome, Pars planitis, Pemphigus vulgaris, Pernicious anaemia, Perivenous encephalomyelitis, POEMS syndrome, Polyarteritis nodosa, Polymyalgia rheumatica, Polymyositis, Primary biliary cirrhosis, Primary sclerosing cholangitis, Progressive inflammatory neuropathy, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum, Pure red cell aplasia, Rasmussen’s encephalitis, Raynaud phenomenon, Relapsing polychondritis, Reiter’s syndrome, Restless leg syndrome, Retroperitoneal fibrosis, Rheumatoid arthritis, Rheumatic fever, Sarcoidosis, Schizophrenia, Schmidt syndrome, Schnitzler syndrome, Scleritis, Scleroderma, Serum Sickness, Sjogren’s syndrome, Spondyloarthropathy, Still’s disease, Stiff person syndrome, Subacute bacterial endocarditis (SBE), Susac’s syndrome, Sweet’s syndrome, Sydenham chorea, Sympathetic ophthalmia, Systemic lupus erythematosis, Takayasu’s arteritis, Temporal arteritis, Thrombocytopenia, Tolosa-Hunt syndrome, Transverse myelitis, Ulcerative colitis, Undifferentiated connective tissue disease, Undifferentiated spondyloarthropathy, Urticarial vasculitis, Vasculitis, Vitiligo, and Wegener’s granulomatosis.

[0217] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of a condition selected among acute or chronic inflammation.

[0218] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of a condition associated with inflammation.

[0219] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of a condition selected among Acne vulgaris, Allergy, Allergic rhinitis, Asthma, Atherosclerosis, Autoimmune disease, Celiac disease, Chronic prostatitis, Glomerulonephritis, Hypersensitivities, Inflammatory bowel diseases, Pelvic inflammatory disease, Reperfusion injury, Rheumatoid arthritis, Sarcoidosis, Transplant rejection, Vasculitis, interstitial cystitis, Cancer, Depression, Myopathies, and Leukocyte defects.

[0220] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of Sepsis.

[0221] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of asthma.

[0222] According to an embodiment, the invention concerns the use of an immune suppressive domain for the manufacture of a medicament for prophylaxis or treatment of allergy.

[0223] According to an embodiment, the invention concerns a method for the preparation of a pharmaceutical composition comprising the steps of: [0224] a. Providing one or more immunosuppressive peptides selected from Seqid 1 to Seqid 287, and optionally cross-linking said one or more immunosuppressive peptides; [0225] b. Optionally providing a carrier; [0226] c. Providing a substance; [0227] d. Mixing the provided one or more peptides with any carrier of optional step b. and the substance of step d. to obtain the pharmaceutical composition.

[0228] According to an embodiment, the invention concerns the method, wherein said substance of step c. is selected from the group consisting of creams, lotions, shake lotions, ointments, gels, balms, salves, oils, foams, shampoos, sprays, aerosols, transdermal patches and bandages.

[0229] According to an embodiment, the invention concerns a pharmaceutical composition obtainable according to the invention.

[0230] According to an embodiment, the invention concerns a pharmaceutical composition comprising an immune suppressive domain as an active substance.

[0231] According to an embodiment, the invention concerns the pharmaceutical composition, wherein said immune suppressive domain is selected among the immune suppressive domains of the invention.

[0232] According to an embodiment, the invention concerns the pharmaceutical composition, further comprising at least one carrier.

[0233] According to an embodiment, the invention concerns the pharmaceutical composition, wherein said at least one carrier is a non-protein carrier and/or a protein carrier.

[0234] According to an embodiment, the invention concerns a use of the composition according to the invention, for treatment of a disease by IV injection.

[0235] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a disease by direct injection at a site of inflammation.

[0236] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a disease by inhalation.

[0237] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a disease by oral delivery.

[0238] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a disease by anal delivery.

[0239] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a condition selected among a skin disease, Psoriasis, Arthritis, Asthma, Sepsis and inflammatory bowel disease.

[0240] According to an embodiment, the invention concerns the use of a composition of the invention for administration in a way selected among IV, IP, and IM.

[0241] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of Arthritis wherein the composition is injected directly at site of inflammation.

[0242] According to an embodiment, the invention concerns the use of a composition of the invention for treatment of a condition selected among Gastrointestinal hyperresponsiveness, Food Allergy, Food intolerance and inflammatory bowel disease, wherein the composition is delivered orally.

[0243] According to an embodiment, the invention concerns the use of a composition of the invention for treatment Asthma where the composition is delivered by inhalation.

[0244] According to an embodiment, the invention concerns the use of a composition of the invention for coating of nanoparticles and biomaterials. The immune suppressive domain may aid in suppressing any immune response e.g. from a patient treated with or subjected to nanoparticles, e.g. for drug delivery or diagnostics, or biomaterials.

[0245] According to an embodiment, the invention concerns the use of a composition of the invention to aid in suppressing any immune response to nanoparticles or biomaterials.

[0246] According to an embodiment, the invention concerns the use of a composition of the invention to increase the half-life of nanoparticles or biomaterials in vivo in a patient.

[0247] According to an embodiment, the invention concerns a vaccine comprising an immune suppressive domain, optionally mutated, for systemic immune suppression.

[0248] According to an embodiment, the invention concerns a vaccine comprising an immune suppressive domain selected among Seqid 275 to 287 against PRRS.

[0249] According to an embodiment, the invention concerns a vaccine comprising a peptide, obtained by performing at least one mutation, insertion or deletion of an immune suppressive domain selected among Seqid 275 to 287.

[0250] According to an embodiment, the invention concerns a vaccine against PRRS comprising a mutated immunosuppressive domain selected among seqid 275 to seqid 287, subject to the proviso that the immunosuppressive properties of said domain have been abrogated.

[0251] According to an embodiment, the invention concerns a peptide having the sequence of the Immune Suppressive Domain according to the invention.

[0252] According to an embodiment, the invention concerns the peptide having the sequence of the Immune Suppressive Domain according to the invention, modified by a number of point mutations selected among 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0253] According to an embodiment, the invention concerns the peptide having the sequence of the Immune Suppressive Domain according to the invention, modified by a number of point deletions selected among 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0254] According to an embodiment, the invention concerns the peptide having the sequence of the Immune Suppressive Domain according to the invention, modified by a number of point insertions selected among 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0255] According to an embodiment, the invention concerns the peptide according to the invention, capable of an inhibition selected among at least 5% inhibition of T-lymphocyte proliferation, at least 10%, at least 20%, such as at least 30%, at least 40%, at least 50%, such as at least 60%, such as at least 70% inhibition of T-lymphocyte proliferation, at least 75% inhibition of T-lymphocyte proliferation, at least 80%, such as at least 85%, at least 90%, such as at least 95%, at least 97%, such as at least 99%, and at least 100% inhibition of T-lymphocyte proliferation.

[0256] According to an embodiment, the invention concerns a peptide according to the invention, capable of suppressing the immune response in an animal, preferably according to the TPA model.

[0257] According to an embodiment, the invention concerns the use of a peptide according to the invention, for a use selected among any of the uses of the invention.

[0258] According to an embodiment, the invention concerns a method of treatment of an indication selected among the indications of the present application and the viral infections of Table 1 comprising administration of an effective amount of an entity selected among the Immune Suppressive Domains of the invention, the compositions of the invention, and the peptides of the invention.

[0259] The co-pending patent application PCT/DK2012/050381 provides a number of immunosuppressive domains. Table 1 is provided below. Sequences of the table are applicable for the purposes of the present invention. Vira of the table provide examples of relevant indications for the present invention.

TABLE-US-00002 family genus Species (group) Species (Strain) Putative ISU as identified using the criteria described in this application fir udentification of immubosupprssive domains Peptides from domains from fusion proteins exhibiting immunosuppressive activity (ISU) Name of envelope attachment / fusion protein IU ggroup and fusion type Flavi-viridae Flavi-virus Aroa virus Bussuquara virus guape virus Naranjal virus seqid85 NRGWNNGCGLFGKG * * * * * * * * * * * * * * seqid7 GDAAWDFGSVGGVFNSLGK * * o * * * * o * * * * * oo * o * * envelope protein prME fusion pretein E Group 1 Type II Fusion mechanism Dengue virus Dengue 1 seqid8 GGTAWDFGSIGGVFTSVGK *o***************** seqid2 DRGWGNGCGLFGKG ************** seqid172 KGS SIGKMFESTYRGAKRMAILG Dengue 2 seqid9 GDTAWDFGSLGGVFTSVGK ****************o** seqid173 KGS SIGKMFEATARGARRMAILG seqid2 DRGWGNGCGLFGKG ************** Dengue 3 seqid174 KGSSIGQMFETTMRGAKRMAILG seqid2 DRGWGNGCGLFGKG ************** seqid204 GDTAWDFGSVGGVLNSLGK ******************* Dengue 4 seqid10 GETAWDFGSVGGLLTSLGK ************oo***** seqid173 KGSSIGKMFEATARGARRMAILG seqid2 DRGWGNGCGLFGKG ************** Japanese encephalitis virus group Japanese encephalitis virus seqid11 LGDTAWDFGSIGGVFNSIG ***o*************** seqid2 DRGWGNGCGLFGKG ************** Koutango virus seqid12 LGDTAWDFGSVGGIFTSLG seqid2 DRGWGNGCGLFGKG ************** Murray Valley encephalitis virus seqid13 LGDTAWDFGSVGGVFNSIG seqid2 DRGWGNGCGLFGKG ************** St. Louis encephalitis virus seqid11 LGDTAWDFGSIGGVFNSIG ******************* seqid2 DRGWGNGCGLFGKG ************** Usutu virus seqid14 LGDTAWDFGSVGGIFNSVG ******************* seqid2 DRGWGNGCGLFGKG ************** West Nile virus seqid15 LGDTAWDFGSVGGVFTSVG **********o******** seqid2 DRGWGNGCGLFGKG ************** Kokobera virus group Kokobera virus unclassified Kokobera virus group seqid16 IGDDAWDFGSVGGILNSVG seqid2 DRGWGNGCGLFGKG Modoc virus group Modoc virus seqid17 VGSAFWNSDQRFSAINLMD seqid18 DRGWGNGCALFGKG Cowbone Ridge virus Jutiapa virus Sal Vieja virus San Perlita virus mosquito-borne viruses Ilheus virus seqid84 LGDTAWDFGSVGGIFNSIG seqid2 DRGWGNGCGLFGKG Sepik virus seqid19 TGEHSWDFGSTGGFFASVG seqid2 DRGWGNGCGLFGKG Ntaya virus group Bagaza virus seqid20 LGDTAWDFGSVGGFFTSLG seqid2 DRGWGNGCGLFGKG Tembusu virus seqid83 LGDTAWDFGSVGGVLTSIG seqid2 DRGWGNGCGLFGKG Yokose virus seqid21 IGDDAWDFGSTGGIFNTIG seqid2 DRGWGNGCGLFGKG Rio Bravo virus group Apoi virus seqid22 SSAFWNSDEPFHFSNLISII seqid2 DRGWGNGCGLFGKG Entebbe bat virus seqid23 GDDAWDFGSTGGIFNTIGKA seqid2 DRGWGNGCGLFGKG Rio Bravo virus seqid24 SSAYWSSSEPFTSAGIMRIL seqid2 DRGWGNGCGLFGKG Saboya virus seqid18 DRGWGNGCALFGKG seqid25 GSSSWDFSSAGGFFGSIGKA Seaborne tick-borne virus group Meaban virus seqid26 GDAAWDFGSVGGFMTSIGRA seqid27 DRGWGNHCGLFGKG Saumarez Reef virus seqid28 GETAWDFGSAGGFFTSVGRG seqid27 DRGWGNHCGLFGKG Tyuleniy virus seqid29 GEAAWDFGSAGGFFQSVGRG seqid27 DRGWGNHCGLFGKG Spondweni virus group Zika virus seqid30 LGDTAWDFGSVGGVFNSLGK *************oo**o** seqid2 DRGWGNGCGLFGKG Kyasanur forest disease virus seqid31 VGEHAWDFGSVGGMLSSVG *************o***** seqid27 DRGWGNHCGLFGKG Langat virus seqid32 VLGEHAWDFGSVGGVMTSIG seqid27 DRGWGNHCGLFGKG Louping ill virus seqid33 IGEHAWDFGSAGGFFSSIG **********o***oo*o* seqid27 DRGWGNHCGLFGKG Omsk hemorrhagic fever virus seqid34 LGEHAWDFGSTGGFLSSIG seqid27 DRGWGNHCGLFGKG Powassan virus seqid35 VGEHAWDFGSVGGILSSVG *************o***** seqid36 DRGWGNHCGFFGKG ************* Royal Farm virus seqid27 DRGWGNHCGLFGKG Tick-borne encephalitis virus seqid37 IGEHAWDFGSAGGFLSSIG seqid38 IGEHAWDFGSTGGFLTSVG seqid39 IGEHAWDFGSTGGFLASVG seqid27 DRGWGNHCGLFGKG Yaounde virus seqid40 LGDTAWDFGSIGGVFTSLG seqid2 DRGWGNGCGLFGKG Yellow fever virus group Banzi virus seqid41 VGSSSWDFSSTSGFFSSVG seqid2 DRGWGNGCGLFGKG Bouboui virus seqid42 VGRSSWDFSSAGGFFSSVG seqid2 DRGWGNGCGLFGKG Edge Hill virus Uganda S virus Wesselsbron virus Yellow fever virus seqid43 MGDTAWDFSSAGGFFTSVG ***o*************** seqid2 DRGWGNGCGLFGKG unclassified Flavivirus Batu Cave virus Cacipacore virus Calbertado virus Cell fusing agent virus Chaoyang virus Chimeric Tick-borne encephalitis virus/Dengue virus 4 Culex theileri flavivirus Donggang virus Duck hemorrhagic ovaritis virus Flavivirus Aedes/MO-Ac/ITA/2009 Flavivirus Anopheles/PV-Am/ITA/2009 Flavivirus CbaAr4001 Flavivirus FSME Flavivirus Phlebotomine/76/Arrabida/2007 Gadgets Gully virus Greek goat encephalitis virus Jugra virus Kadam virus Kamiti River virus Kedougou virus Montana myotis leukoencephalitis virus Mosquito flavivirus Ngoye virus Nounane virus Phlebotomus flavivirus Alg_F19 Phlebotomus flavivirus Alg_F8 Quang Binh virus Russian Spring-Summer encephalitis virus Sokoluk virus Spanish sheep encephalitis virus T'Ho virus Tai forest virus B31 Tamana bat virus Tick-borne flavivirus Wang Thong virus Flavivirus sp. seqid44 NRGWGTGCFKWGIG seqid45 NRGWGTGCFEWGLG seqid2 DRGWGNGCGLFGKG Aedes flavivirus seqid45 NRGWGTGCFEWGLG seqid46 HVAGRYSKHGMAGIGSVWEDLVR Culex flavivirus seqid44 NRGWGTGCFKWGIG seqid47 VDKYRRFGTAGVGG Hepa civi rus Hepatitis C virus Hepatitis C virus genotype 1 a seqid3 GLIHLHQNIVDVQYLYG seqid175 PALSTGLIHLHQNIVDVQ E1/E2 Hepatitis C virus genotype 1b seqid48 GLIHLHRNIVDVQYLYG seqid176 PALSTGLIHLHRNIVDVQ Hepatitis C virus genotype 2 seqid49 GLIHLHQNIVDVQYMYG seqid175 PALSTGLIHLHQNIVDVQ Hepatitis C virus genotype 3 seqid175 PALSTGLIHLHQNIVDVQ seqid3 GLIHLHQNIVDVQYLYG Hepatitis C virus genotype 4 seqid175 PALSTGLIHLHQNIVDVQ seqid3 GLIHLHQNIVDVQYLYG Hepatitis C virus genotype 5 seqid50 GLIHLHQNIVDTQYLYG seqid177 PALSTGLIHLHQNIVDTQ Hepatitis C virus genotype 6 seqid175 PALSTGLIHLHQNIVDVQ seqid3 GLIHLHQNIVDVQYLYG All Hepatitis C virus seqid3 GLIHLHQNIVDVQYLYG Border disease virus Border disease virus - Border disease virus - X818 Border disease virus 1 Border disease virus 2 Border disease virus 3 Border disease virus isolates seqid51 NTTLLNGSAFQLICPYGWVGRVEC seqid52 SYFQQYMLKGQYQYWFDLE Bovine viral diarrhea virus 1 Bovine viral diarrhea virus 1-CP7 Bovine viral diarrhea virus 1-NADL Bovine viral diarrhea virus 1-Osloss Bovine viral diarrhea virus 1-SD1 Bovine viral diarrhea virus isolates and strains Bovine viral diarrhea virus type 1a Bovine viral diarrhea virus type 1b Pestivirus isolate 97-360 Pestivirus isolate Hay 87/2210 Pestivirus strain mousedeer Pestivirus type 1 isolates seqid53 NTTLLNGPAFQMVCPLGWTGTVSC seqid54 SYFQQYMLKGEYQYWFDLE Bovine viral diarrhea virus 2 (BVDV-2) Bovine viral diarrhea virus 2 Pestivirus sp. strain 178003 Pestivirus sp. strain 5250Giessen-3 Bovine viral diarrhea virus-2 isolate SCP seqid55 SLLNGPAFQMVCPQGWTGTIEC seqid56 DRYFQQYMLKGKWQYWFDLD Classical swine fever virus Classical swine fever virus Hog cholera virus strain Zoelen seqid57 TLLNGSAFYLVCPIGWTGVIEC seqid58 SYFQQYMLKGEYQYWFDLD unclassified Pestivirus Bovine viral diarrhea virus 3 seqid59 TLLNGPAFQLVCPYGWTGTIEC seqid60 DNYFQQYMLKGKYQYWFDLEATD Chamois pestivirus 1 seqid61 TLLNGSAFQMVCPFGWTGQVEC seqid62 DSYFQQYMLKGEYQYWFDLDAKD Porcine pestivirus isolate Bungowannah seqid205 TLLNGPAFQLVCPYGWTGTIECDSYYQQYII KSGYQYWFDLTAKD Unnclassified Flaviviridae Barkedji virus Canine hepacivirus AAK-2011 GB virus A Douroucouli hepatitis GB virus A GBV-A-like agents GB virus D GBV-C/HGV group GB virus C Hepatitis GB virus C-like virus Hepatitis GB virus B Lammi virus Marmoset hepatitis GB virus A Nakiwogo virus Turkey meningoencephalitis virus Togaviridae Alpha-virus Aura virus Barmah Forest virus Middelburg virus Ndumu virus Salmon pancreas disease virus Getah virus Mayaro virus Trocara virus EEEV complex seqid63 GVYPFMWGGAYCFCDTENTQVS **********o****o**o*o* seqid64 APFGCEIYTNPIRAENCAVGSIP *****o*ooo*o**oo*oo*oo* seqid65 SDFGGIATVKYSASKSGKCAVH o***oooooo*ooooo*o*oo* seqid66 FSTANIHPEFRLQICTSYVTCKGDCHPP *oooooooo*oooo*ooooo*ooo*o** WEEV complex Fort Morgan virus Highlands J virus Sindbis virus Western equine encephalomyelitis virus Whataroa virus VEEV complex Cabassou virus Mucambo virus Pixuna virus Venezuelan equine encephalitis virus seqid67 GVYPFMWGGAYCFCD *************** seqid68 GDCHPPKDHIVTHPQYHAQ ************o**o*o* seqid69 AVSKTAWTWLTS *********oo* SFV complex Bebaru virus O'nyong-nyong virus Ross River virus Semliki forest virus Una virus seqid63 GVYPFMWGGAYCFCDTENTQVS **********o****o**o*o* seqid64 APFGCEIYTNPIRAENCAVGSIP *****o*ooo*o**oo*oo*oo* seqid65 SDFGGIATVKYSASKSGKCAVH o***oooooo*ooooo*o*oo* seqid66 FSTANIHPEFRLQICTSYVTCKGDCHPP *oooooooo*oooo*ooooo*ooo*o** Chikungunya virus seqid67 GVYPFMWGGAYCFCD *************** seqid70 VHCAAECHPPKDHIVNY oo*o*o**o******** seqid71 PASHTTLGVQDISATAMSWV o****oo******o****** Rubivirus Rubella virus Rubella virus (strain BRD1) Rubella virus (strain BRDII) Rubella virus (strain Cendehill) Rubella virus (strain M33) Rubella virus (strain RN-UK86) Rubella virus (strain THERIEN) Rubella virus (strain TO-336 vaccine) Rubella virus (strain TO-336) Rubella virus (vaccine strain RA27/3) seqid72 ACTFWAVNAYSSGGYAQLASYFNPGGSYYK ***o*o****o**oo****o**o******o seqid73 QYHPTACEVEPAFGHSDAACWGFPTDT ***o*o*o*o****o********o*** seqid74 MSVFALASYVQHPHKTVRVKFHT ***oo*****o**o**o****** seqid159 ETRTVWQLSVAGVSC o*o* ********oo* seqid76 NVTTEHPFCNMPHGQLEVQVPP o*o*o**oo*o*o****o*oo* seqid77 DPGDLVEYIMNYTGNQQSRW ****o******o*o****** seqid78 GSPNCHGPDWASPVCQRHSPDCS ****o***o************** seqid79 RLVGATPERPRLRLV o***o**o**o**** seqid80 DADDPLLRTAPGP *oo********** seqid81 GEVWVTPVIGSQARKCGL oo*o**o**o*****o** seqid86 HIRAGPYGHATVEM oo***********o seqid87 PEWIHAHTTSDPWHP o**oooo*o***o*o seqid88 PGPLGLKFKTVRPVALPR ****o***o**o*oo*** seqid89 ALAPPRNVRVTGCYQCGTPAL oooo**o*o*o**o******* seqid90 EGLAPGGGNCHLTVNGEDVG ***o*****o**oo*o*oo* seqid207 LLNTPPPYQVSCGG ******o*o*o*** 46 seqid92 RASARVIDPAAQSFTGWYGTHT **o***oo*o************* seqid93 TAVSETRQTWAEWAAAHWWQLTLG o*******ooo*****o******* Bunya-viridae Hanta-virus (continued on next page) Amur virus Bayou virus Black Creek Canal virus Cano Delgadito virus Calabazo virus Catacamas virus Choclo virus Dobrava-Belgrade virus El Moro Canyon virus Hantaan virus Isla Vista virus Khabarovsk virus Laguna Negra virus Limestone Canyon virus Monongahela virus Muleshoe virus Muju virus New York virus Oran virus Playa de Oro virus Prospect Hill virus Puumala virus Rio Mamore virus Rio Segundo virus Saaremaa virus Seoul virus Sin Nombre virus Soochong virus Thailand virus Thottapalayam virus Topografov virus Tula virus seqid94 NPPDCPGVGTGCTACGVYLD **o****o********o*** seqid95 RKVCIQLGTEQTCKTIDSNDC *oo*o*o*o*oo**oo*o*** seqid96 DTLLFLGPLEEGGMIFKQWCTTTCQFGDPGD IM seqid97 GSFRKKCSFATLPSCQYDGNTVSG *o***o*o***o*o*ooo**oo** seqid98 ATKDSFQSFNITEPH **o****o**oooo* seqid99 GSGVGFNLVCSVSLTEC ******o*o*ooo**** seqid100 KACDSAMCYGSSTANLVRGQNT ****o*o***ooooo*o**o** seqid101 GKGGHSGSKFMCCHDKKCSATGLVAAAPHL ********o*o***ooo*ooo**o*oo*** seqid102 DDGAPQCGVHCWFKKSGEW ***o*o*ooo***oo**** Gn(G2)/Gc(G1) Ortho-bunya-virus Anopheles A virus Anopheles B virus Bakau virus Batama virus Bwamba virus Caraparu virus Kaeng Khoi virus Kairi virus Madrid virus Main Drain virus Marituba virus Nyando virus Oriboca virus Oropouche virus Sathuperi virus Shamonda virus Shuni virus Simbu virus Tacaiuma virus Tete virus Turlock virus unclassified Orthobunyavirus seqid103 KHDELCTGPCPVNINHQTGWLT *o*o***o**oooooooo*o*o seqid104 WGCEEFGCLAVSDGCVFGSCQD **o*oo**o*ooo**oo***** seqid105 GNGVPRFDYLCHLASRKEVIVRKC *o*ooo*ooo*oooo*ooooo*o* seqid106 SCAGCINCFQNIHC *o**ooooooooo* Akabane virus Sabo virus Tinaroo virus Yaba-7 virus Bunyamwera virus Batai virus Birao virus Bozo virus Cache Valley virus Fort Sherman virus Germiston virus Guaroa virus Iaco virus Ilesha virus Lokern virus Maguari virus Mboke virus Ngari virus Northway virus Playas virus Potosi virus Shokwe virus Tensaw virus Tlacotalpan virus Xingu virus California Encephalitis virus California encephalitis serogroup virus LEIV California encephalitis virus - BFS-283 Chatanga virus Inkoo virus Jamestown Canyon virus Jamestown Canyon-like virus Jerry Slough virus Keystone virus La Crosse virus Lumbo virus Melao virus Morro Bay virus San Angelo virus Serra do Navio virus Snowshoe hare virus South River virus Tahyna virus Trivittatus virus Caraparu virus Apeu virus Bruconha virus Ossa virus Vinces virus Manzanilla virus Buttonwillow virus Ingwavuma virus Mermet virus Marituba virus Gumbo Limbo virus Murutucu virus Nepuyo virus Restan virus Wyeomyia virus Anhembi virus BeAr328208 virus Macaua virus Sororoca virus Taiassui virus Phlebovirus Bujaru virus Candiruvirus Chilibre virus Frijoles virus Punta Tor.sub.└Salehabad virus Sandflyfever Naples virus Uukuniemi viruso virus Rift Valley fever virus seqid107 KTVSSELSCREGQSYWT **oo**oo*o**o*o** seqidl08 GSFSPKCLSSRRC *******oooooo seqid109 ENKCFEQCGGWGCGCFNVNPSCLFVHT **o*o**o*oo*oo***ooo***o**o seqid110 WGSVSLSLDAEGISGSNSFSF **ooo*o**o*o*o*o*oo** seqid111 RQGFLGEIRCNSE *o*****o**oo* seqid112 AHESCLRAPNLVSYKPMIDQLEC *oo**oo**oooo*o*oo*ooo* seqid113 DPFWFERGSLPQTR **ooo*oo*o***o* seqid114 QAFSKGSVQADLTLMFD **ooo*ooo*oooooo* seqidll5 CDAAFLNLTGCYSCNAG *o*o*o*oo*****oo* seqid116 CQILHFTVPEVEEEFMYSC *ooo*ooo*ooooooo*o* seqid117 STVVNPKSGSWN *o*o**oooooo seqid118 FFDWFSGLMSWFGGPLK *o***oo*o**oooooo Unclassified Phelobvirus (continued on next page) Anhanga virus Arumowot virus Chagres virus Corfou virus Gabek Forest virus Itaporanga virus Phlebovirus Adria/ALB1/2005 Phlebovirus Adria/ALB5/2005 Phlebovirus AH12 Phlebovirus AH12/China/2010 Phlebovirus AH15/China/2010 Phlebovirus B105-05 Phlebovirus B151-04 Phlebovirus B43-02 Phlebovirus B68-03 Phlebovirus B79-02 Phlebovirus Chios-A Phlebovirus Cyprus Phlebovirus HB29/China/2010 Phlebovirus HN13/China/2010 Phlebovirus HN6/China/2010 Phlebovirus Hu/Xinyang1/China/2010 Phlebovirus Hu/Xinyang2/China/2010 Phlebovirus IB13-04 Phlebovirus JS2007-01 Phlebovirus JS24 Phlebovirus JS26 Phlebovirus JS3/China/2010 Phlebovirus JS4/China/2010 Phlebovirus JS6 Phlebovirus JSD1 Phlebovirus LN2/China/2010 Phlebovirus LN3/China/2010 Phlebovirus sandflies/Gr29/Spain/2004 Phlebovirus sandflies/Gr36/Spain/2004 Phlebovirus sandflies/Gr44/Spain/2004 Phlebovirus sandflies/Gr49/Spain/2004 Phlebovirus sandflies/Gr52/Spain/2004 Phlebovirus sandflies/Gr65/Spain/2004 Phlebovirus sandflies/Gr98/Spain/2004 Phlebovirus SD24/China/2010 Phlebovirus SD4/China/2010 Phlebovirus tick/XCQ-2011 Phlebovirus XLL/China/2009 Rio Grande virus Salobo virus Sandfly fever sicilian virus Sandfly Sicilian Turkey virus Utique virus Phlebovirus sp. Phlebovirus sp. Be An 24262 Phlebovirus sp. Be An 356637 Phlebovirus sp. Be An 416992 Phlebovirus sp. Be An 578142 Phlebovirus sp. Be Ar 371637 Phlebovirus sp. Co Ar 170255 Phlebovirus sp. Co Ar 171616 Phlebovirus sp. GML 902878 Phlebovirus sp. Pa Ar 2381 Phlebovirus sp. PAN 479603 Phlebovirus sp. PAN 483391 Phlebovirus sp. VP-161A Phlebovirus sp. VP-334K Phlebovirus sp. VP-366G Orthomyxoviridae Influenzavirus A Influenza A virus INFA H1 seqid119 GLFGAIAGFIEGGWTG seqid178 WTYNAELLVLLENERTLD seqid179 NAELLVLLENERTLDYHD INF F#2 DELTA6: seqid201 GLFGAAGFIENGWEG InFAHI-3: seq1d203 HA (Ha1/HA2) INFA H2 seqid120 GLFGAIAGFIEGGWQG seqid180 WTYNAELLVLMENERTLD seqid181 NAELLVLMENERTLDYHD INFA H3 seqid121 GIFGAIAGFIENGWEG seqid182 WSYNAELLVALENQHTID seqid183 NAELLVALENQHTIDLTD seqid4 GLFGAIAGFIENGWEG INFA H4 seqid122 GLFGAIAGFIENGWQG seqid182 WSYNAELLVALENQHTID seqid184 NAELLVALENQHTIDVTD INFA H5 seqid120 GLFGAIAGFIEGGWQG seqid180 WTYNAELLVLMENERTLD seqidl85 NAELLVLMENERTLDFHD INFA H6 seqid123 GIFGAIAGFIEGGWTG seqid119 GLFGAIAGFIEGGWTG seqid178 WTYNAELLVLLENERTLD seqid186 NAELLVLLENERTLDMHD INFA H7 seqid187 WSYNAELLVAMENQHTID seqid208 WSYNAELLVAMENQHLAD seqid4 GLFGAIAGFIENGWEG INFA H8 seqid124 GLFGAIAGFIEGGWSG seqid189 WAYNAELLVLLENQKTLD seqid190 NAELLVLLENQKTLDEHD INFA H9 seqid125 GLFGAIAGFIEGGWPG seqid124 GLFGAIAGFIEGGWSG seqid189 WAYNAELLVLLENQKTLD seqid190 NAELLVLLENQKTLDEHD INFA H10 seqid191 WTYQAELLVAMENQHTID seqid192 QAELLVAMENQHTIDMAD seqid4 GLFGAIAGFIENGWEG INFA H11 seqid125 GLFGAIAGFIEGGWPG seqid193 WSYNAQLLVLLENEKTLD seqid194 NAQLLVLLENEKTLDLHD INFA H12 seqid125 GLFGAIAGFIEGGWPG seqid189 WAYNAELLVLLENQKTLD seqid190 NAELLVLLENQKTLDEHD INFA H13 seqid125 GLFGAIAGFIEGGWPG seqid195 WSYNAKLLVLLENDKTLD seqid196 NAKLLVLLENDKTLDMHD INFA H14 seqid122 GLFGAIAGFIENGWQG seqid182 WSYNAELLVALENQHTID seqid184 NAELLVALENQHTIDVTD INFA H15 seqid187 WSYNAELLVAMENQHTID seqid188 NAELLVAMENQHTIDLAD seqid4 GLFGAIAGFIENGWEG INFA H16 seqid125 GLFGAIAGFIEGGWPG seqid197 WSYNAKLLVLIENDRTLD seqid198 NAKLLVLIENDRTLDLHD Influenzavirus B Influenza B virus All strains seqid126 GFFGAIAGFLEGGWEG seqid199 ISSQIELAVLLSNEGIIN seqid200 QIELAVLLSNEGIINSED Influenzavirus C Influenza C virus Paramyxoviridae Avulavirus Avian paramyxovirus 2 Yucaipa virus Avian paramyxovirus 3 Avian paramyxovirus 3b Avian paramyxovirus 4 Avian paramyxovirus 5 Avian paramyxovirus 6 Avian paramyxovirus 7 Avian paramyxovirus 8 Avian paramyxovirus 9 Newcastle disease virus Pigeon paramyxovirus 1 unclassified Avulavirus Avian paramyxovirus 10_Avian paramyxovirus duck/Miyagi/885/05 Avian paramyxovirus penguin/Falkland Islands/324/2007 Goosramyxovirus HZ Goose paramyxovirus JS/1/97/Go Goose paramyxovirus SF02 seqid127 GAIALGVATAAAVTAG oooo*o*oo*o*oo** F0 (F2/F1) Henipavirus Hendra virus Hendra virus horse/Australia/Hendra/1994 Nipah virus unclassified Henipavirus Bat paramyxovirus Eid.hel/GH45/2008 Morbillivirus Canine distemper virus Cetacean morbillivirus_Dolphin morbillivirus_Pilot whale morbillivirus Porpoise morbillivirus Measles virus Peste-des-petits-ruminants virus Phocine distemper virus Phocine distemper virus 1 Phocine distemper virus-2 Rinderpest virus Respirovirus Bovine parainfluenza virus 3 Porcine paramyxovirus strain Frost Porcine paramyxovirus strain Texas Human parainfluenza virus 1 Human parainfluenza virus 3 Simian Agent 10 Sendai virus unclassified Respirovirus Atlantic salmon respirovirus Guinea pig parainfluenza virus TS-9 Pacific salmon paramyxovirus Trask River 1983 Swine parainfluenza virus 3 Tursiops truncatus parainfluenza virus 1 Rubulavirus Human parainfluenza virus 2 Human parainfluenza virus 2 (strain Greer) Human parainfluenza virus 2 (strain Toshiba) Human parainfluenza virus 4 Human parainfluenza virus 4a Human parainfluenza virus 4b Mapuera virus Mumps virus Parainfluenza virus 5 Porcine rubulavirus Simian virus 41 unclassified Rubulavirus Porcine parainfluenza virus Tuhoko virus 1 Tuhoko virus 2 Tuhoko virus 3 unclassified Paramyxovirinae Atlantic salmon paramyxovirus Beilong virus Canine parainfluenza virus Chimeric human parainfluenza virus rPIV3-2 Fer-de-lance virus J-virus Menangle virus Mossman virus Murayama virus Ovine parainfluenza virus 3 Pacific salmon paramyxovirus Paramyxovirus GonoGER85 Recombinant PIV3/PIV1 virus Reptilian paramyxovirus Salem virus Salmo salar paramyxovirus Snake ATCC-VR-1408 paramyxovirus Snake ATCC-VR-1409 paramyxovirus Tioman virus Tupaia paramyxovirus Pneumovirus Human respiratory syncytial virus Human respiratory syncytial virus A Human respiratory syncytial virus (strain RSB1734) Human respiratory syncytial virus (strain RSB5857) Human respiratory syncytial virus (strain RSB6190) Human respiratory syncytial virus (strain RSB6256) Human respiratory syncytial virus (strain RSB642) Human respiratory syncytial virus (strain RSB6614) Human respiratory syncytial virus A strain Long LinkOut Human respiratory syncytial virus A2 Human respiratory syncytial virus B Human respiratory syncytial virus (subgroup B / strain 18537) Human respiratory syncytial virus (subgroup B / strain 8/60) Human Respiratory syncytial virus 9320 Human respiratory syncytial virus B1 Human respiratory syncytial virus S2 Human respiratory syncytial virus strain RSS-2 unclassified Human respiratory syncytial virus seqid128 FLGLILGLGAAVTAGVA ***oo**o*o*ooo*o* seqid129 TNEAWSLTNGMSVL **o*****o**o*** seqid130 VIRFQQLNKRLLE **o***o*o**** seqid131 REFSSNAGLT ****o***o* seqid132 MLTDRELTSIVGGM ***oo**o*oooo* seqid133 YVIQLPLFGVMDTDCW *oo***oo**o**o** seqid134 CLARADNGWYCHNAGSLSYFP **ooo*o**o*o****o*o** seqid135 DTLKSLTVPVTSRECN **oo***o*ooooo** seqid136 YDCKISTSKTYVSTAVLTTMG *o*o*o***ooo*oo*o*oo* seqid137 VSCYGHNSCTVIN *****ooo**oo* Seqid75 GIIRTLPDGCHYISNKGVDRVQVGNTVYYLS KEVGK ***o*ooo**o*o**o*o*o*o****o**oo *oo** seqid139 PLSFPDDKFDVAIRDVEHSINQTRTFLKASD QLL **o**o*o*ooo*oo*ooo***ooo*ooo** o** seqid140 KIMTSKTDISSSVITSIGAIVSCYG o*o***ooo*oo*o*oo*oo***** Bovine respiratory syncytial virus All strains seqid128 FLGLILGLGAAVTAGVA ***oo**o*o*ooo*o* seqid134 CLARADNGWYCHNAGSLSYFP **ooo*o**o*o****o*o** seqid133 YVIQLPLFGVMDTDCW *oo***oo**o**o** Metapneumovirus Avian metapneumo-virus All strains Human metapneumo-virus All strains Coronaviridae Coronavirinae Alphacoronavirus Alphacoronavirus 1 Coronavirus group 1b Human coronavirus 229E Human coronavirus NL63 Miniopterus bat coronavirus 1 Miniopterus bat coronavirus HKU8 Porcine epidemic diarrhea virus Rhinolophus bat coronavirus HKU2 Scotophilus bat coronavirus 512 unclassified Alphacoronavirus S (S1/S2) Betacoronavirus Betacoronavirus 1 Coronavirus group 2b Coronavirus group 2c Human coronavirus HKU1 Murine coronavirus Pipistrellus bat coronavirus HKU5 Rousettus bat coronavirus HKU9 Severe acute respiratory syndrome-related coronavirus recombinant SARSr-CoV SARS coronavirus Tylonycteris bat coronavirus HKU4 unclassified Betacoronavirus Gammacorona-virus Avian coronavirus Beluga Whale coronavirus SW1 unclassified coronaviruses Alpaca coronavirus CA08-1/2008 Bat coronavirus Bird droppings coronavirus Bovine respiratory coronavirus Chicken enteric coronavirus Coronavirus Anas Coronavirus oystercatcher/p17/2006/GBR Coronavirus red knot/p60/2006/GBR Ferret enteric coronavirus 1202 Ferret systemic coronavirus MSU-S Ferret systemic coronavirus WADL Guangxi coronaviridae Human coronavirus NO Human enteric coronavirus strain 4408 Kenya bat coronavirus Mink coronavirus strain WD1133 Parrot coronavirus AV71/99 Quail coronavirus Italy/Elvia/2005 Tai Forest coronavirus unidentified coronavirus unidentified human coronavirus seqid141 RSAIEDLLFDKVKLSDVG **oo****oo**ooo*o* seqid142 VPFYLNVQYRINGLGVT 0**00000**0**0*** seqid143 VLSQNQKLIANAFNNALHAIQ **00***0*000*00*000** seqid144 TNSALVKIQAVVNANA *oo**o*o*o***oo* seqid145 AEAQIDRLINGRLTALNAYVSQQL *oo******o***oo*oo*oo*** seqid146 SAAQAMEKVNECVKSQSSRINFCGNGNHIIS 0*00*00*00***00*00*00***0*0*00* seqid147 APYGLYFIHFNYVP **o*oo*o*oo*o* seqid148 LQEAIKVLNHSYINLKDIGTYEYYVKWPWYV W 00*00*0**0*000*000*00*0*0*****0 * seqid209 EVFAQVKQMYKT PTLKYFGGFNFSQIL seqid210 EVFAQVKQMYKT PAIKDFGGFNFSQIL Segid211 SFIEDLLFNKVTLADAGF Segid212 SAIEDLLFNKVRLSDVGF Seqid213 SLLEDLLFNKVKLSDVGF Seqid214 SAIEDLLFSKVKLADVGF Seqid215 SAIEDLLFDKVKLSDVGF Arena-viridae Arean-virus LCMV-Lassa virus (Old World) complex Ippy virus Lassa virus Lujo virus Lymphocytic choriomeningitis virus Mobala virus Mopeia virus seqid149 NALINDQLIMKNHLRDIMGIPYC *o**o***o*o***o*o**o*** seqid150 FTWTLSDSEGKDTPGGYCLT 00*000*00*000***0**0 seqid151 KCFGNTAIAKCNQKHDEEFCDMLRLFDFN ***0*000****00*00****000*000* seqid152 MLQKEYMERQGKTPLGLVDLFVFS *000*00**00**00*0*0000*0 GpC (Gp1/Gp2) Tacaribe virus (New World) complex Amapari virus Chapare virus Flexal virus Guanarito virus Junin virus Latino virus Machupo virus Oliveros virus Paraná virus Pichinde virus Pirital virus Sabiá virus Tacaribe virus Tamiami virus Whitewater Arroyo virus seqid150 FTWTLSDSEGKDTPGGYCLT 00*000*00*000***0**0 seqid151 KCFGNTAIAKCNQKHDEEFCDMLRLFDFN ***o*ooo****oo*oo****ooo*ooo* seqid152 MLQKEYMERQGKTPLGLVDLFVFS *000*00**00**00*0*0000*0 Hepadnaviridae Genus Orthohepadnavirus Hepatitis B virus HBV genotype A HBV genotype B HBV genotype C HBV genotype D HBV genotype E HBV genotype F HBV genotype G HBV genotype H Hepatitis B virus alphal L and M and S Where S mediates Hepatitis B virus LSH/chimpanzee Hepatitis B virus strain cpz Hepatitis B virus subtype adr Hepatitis B virus subtype adw Hepatitis B virus subtype adyw Hepatitis B virus subtype ayw seqid153 FNPLGFFPSHQLDPLF o***o*o*o*o*o*o* seqid154 ADWDKNPNKDPWP o*o*o*oo*oooo seqid155 MESITSGFLGPLLVLQAVFF oooooooo*ooooo**oooo seqid156 LLTRILTIPQSLDSWWTSLNFLGGA oooooo*oooo*oooo***o*o*oo seqid157 CPPTCPGYRWMC oo*o*****o*o seqid158 LFILLLCLIFLLVLLDYQ *oo*ooo*oo*oo*oooo Rhabdoviridae Dimarhabdovirus Ephemerovirus Bovine ephemeral fever virus seqid160 LDGYLCRKQKWEVTCTETWYFVTD *o*oo****o*ooo*o*****o*o seqid161 KYQIIEVIPTENEC o***o**o*oooo* seqid162 LKGEYIPPYYPPTNCVWNAIDTQE oo*oo*******oo*o**oooo** seqid163 IEDPVTMTLMDSKFTKPC ooo*oooooo**o*oo** seqid164 LHCQIKSWECIPV o**oo*o****o* seqid165 SHRNMMEALYLESPD *oo*oo*o*oo*o** seqid166 LTFCGYNGILLDNGEWWSIY o****oo**oooo****** seqid167 ELEHEKCLGTLEKLQNGE *****o**o*oo*oo*o* seqid168 LDLSYLSPSNPGKHYAY **o***o*oo**oo*** seqid169 IRAVCYYHTFSMNLD o**o*o*oo*oooo* Glyoprotein G Vesiculovirus Carajas virus Chandipura virus Cocal virus Isfahan virus Maraba virus Piry virus recombinant Vesiculovirus Spring viraemia of carp virus Vesicular stomatitis Alagoas virus Vesicular stomatitis Indiana virus Vesicular stomatitis New Jersey virus seqid170 EWKTTCDYRWYGPQYITHSI o*o****o*****o*o*o* seqid171 LGFPPQSCGWASVTT o****oo**oooooo seqid1 VQVTPHHVLVDEYTGEWVDSQFINGKC ooooo*o*oooo*o*o*o*oooooooo Lyssavirus Aravan virus Australian bat lyssavirus Duvenhage virus European bat lyssavirus 1 European bat lyssavirus 2 Irkut virus Khujand virus Lagos bat virus Mokola virus West Caucasian bat virus Rabies virus Rabies virus AB21 Rabies virus AB22 Rabies virus AVO1 Rabies virus BNG4 Rabies virus BNG5 Rabies virus China/DRV Rabies virus China/MRV Rabies virus CVS-11 Rabies virus ERA Rabies virus Eth2003 Rabies virus HEP-FLURY Rabies virus India Rabies virus Nishigahara RCEH Rabies virus Ontario fox Rabies virus Ontario skunk Rabies virus PM Rabies virus red fox/08RS-1981/Udine/2008 Rabies virus SAD B19 Rabies virus silver-haired bat-associated SHBRV Rabies virus strain Pasteur vaccin Rabies virus strain Street Rabies virus vnukovo-32 Thailand genotype 1 dog lyssavirus seqid5 GFTCTGVVTEAETYTNFVGYVT *o****o**o*oo*oooo*** seqid6 SLHNPYPDYRWLRTVKTT *ooooooooooo***o* Seqid138 ESLVIISPSVADLDPYDRSLHS *ooo***oooo*o**ooo Seqid91 CKLKLCGVLGLRLMDGT *ooo****oooo*ooo* Seqid206 ILGPDGNVLIPEMQSS o**o*ooo*******o seqid82 QHMELLESSVIPLVHPL *ooo**o*ooo**oo** unclassified Lyssavirus Bokeloh bat lyssavirus European bat lyssavirus Lyssavirus Ozernoe Shimoni bat virus Novirhabdovirus Hirame rhabdovirus Infectious hematopoietic necrosis virus Snakehead rhabdovirus Viral hemorrhagic septicemia virus unassigned Rhabdoviridae Bangoran virus Bimbo virus Bivens Arm virus Flanders virus Garba virus Klamath virus Malpais Spring virus Nasoule virus Ngaingan virus Ouango virus Sigma virus Tupaia virus Wongabel virus Filoviridae Lloviu virus (LLOV) Bundibugyo virus (BDBV; previously BEBOV) Reston virus (RESTV; previously REBOV) Sudan virus (SUDV; previously SEBOV) Tai Forest virus (TAFV; previously CIEBOV) Ebola virus (EBOV; previously ZEBOV) Marburg virus (MARV) Ravn virus (RAW) Seqid216 GAALGLAWIPYFGPAAE oo*o*oo***o***ooo seqid217 GAAVGLAWIPYFGPAAE Seqid218 GAAAGLAWIPYFGPAAE Seqid219 DLAAGLSWIPFFGPGIE Seqid220 HNAAGIAWIPYFGPGAE Lentivirisae Hiv1 Seqid221 AVGLGALFLGFLGAAGSTMGAAS oooo**ooo*o*oo*****o**o seqid222 LTLTGQARQLLS o***o*o*o*oo seqid223 GIVQQQSNLLQAIEAQQ o*****o***o*****o seqid224 GLGAMFLGFLGAAGSTMGAASLTLTVQARQL LS Seqid225 GIGAMFLGLLSAAGSTMSAAAITLTVQTRQL LS Seqid226 GIGAMFLGLLSAAGSTMGAAAITLTVQTRQL LS Seqid227 GIGAVFLGFLGAAGSTMGAASITLTVQARQL LS Seqid228 GVGALFLGFLSAAGSTMGAASITLTVQARQL LS Seqid229 GIGAMILGFLGAAGSTMGAASITLTVQARQL LS Seqid230 GLGAMFLGFLGAAGSTMGAASITLTVQARQL LS Seqid231 GFGAMFLGFLGAAGSTMGAASITLTVQARQL LS Seqid232 TLGAMFLGFLGAAGSTMGAASMTLTVQARQL LS Seqid233 GLGAVFLGFLGAAGSTMGAASITLTVQARQL LS Seqid234 TIGAMFLGFLGAAGSTMGAASITLTVQARRL LS Seqid235 TIGAMFLGFLGAAGSTMGAASMTLTVQARLL LS Seqid236 TLGAMFLGFLGAAGSTMGAASMTLTVQARLL FS Seqid237 TLGAMFLGFLGAAGSTMGAASLTLTVQARLL LS Seqid238 GVGAMFLGFLGAAGSTMGAASLTLTVQARQL LS Seqid239 GLGAMFLGFLGAAGSTMGAASITLTVQARLL LS Seqid240 TLGAVFLGFLGAAGSTMGAASLTLTVQARLL LS Seqid241 GIGAVFLGFLGAAGSTMGAASITLTVQARKL LS Seqid242 GIGALFLGFLGAAGSTMGAASVTLTVQARQL LS Seqid243 GLGALFLGFLGAAGSTMGAASVTLTVQARQL LS Seqid244 GIGAMFLGFLGAAGSTMGAASITLTVQARLL LS Seqid245 GIGAMFLGFLGAAGSTMGAASVTLTVQARLL LS Seqid246 AIGALFLGFLGAAGSTMGAASVTLTVQARLL LS Seqid247 TLGAMFLGFLGAAGSTMGAASLTLTVQARQL LS Seqid248 GIGALFLGFLGAAGSTMGAASMTLTVQARQL LS Seqid249 GIGAMFLGFLGAAGSTMGAASLTLTVQARQL LS Seqid250 GIGAVFLGFLGAAGSTMGAASMTLTVQARLL LS Seqid251 GIGALFLGFLGAAGSTMGAASLTLTVQARQL LS Seqid252 GIGAVFLGILGAAGSTMGAASITLTVQARQL LS Seqid253 GIGAVFLGFLGAAGSTMGAASVTLTVQARQL LF Seqid254 GLGAMFFGFLGAAGSTMGAASVTLTVQARQL LS Seqid255 GIGALFLGFLGAAGSTMGAASITLTVQARLL LS Seqid256 GLGALFVGFLGAAGSTMGAASITLTVQARQL LS Seqid257 GIGALFLGFLGTAGSTMGAASVTLTVQARQL LS Seqid258 GIGAMIFGFLGAAGSTMGAASITLTVQARQL LS Seqid259 GLGAVLLGFLGTAGSTMGAASLTLTVQVRQL LS Seqid260 GIGAVLFGFLGAAGSTMGAASITLTVQVRQL LS Seqid261 GLGALFLGFLGAAGSTMGAASLTLTGQARQL LS oo**ooo*o*oo*****o**oo***o*o*o* oo Seqid262 GTLGAMFLGFLGAAGSTMGAASMTLTVQARQ LL Seqid263 GTIGAMFLGFLGAAGSTMGAASITLTVQARR LL Seqid264 GTIGAMFLGFLGAAGSTMGAASMTLTVQARL LL Seqid265 IGALFLGFLGAAGSTMGAASVTLTVQARLLL SG Bovine lentivirus group Seqid266 AVGMVIFLLVLAIMAMTASVTAA ***oo**********o*o*oo** Equine lentivirus group Seqid267 FGISAIVAAIVAATAIAASA **o*ooo**********o*o Feline lentivirus group Seqid268 TLALVTATTAGLIGTTTGTSA Seqid269 HVMLALATVLSMAGAGTGATA Ovine/caprine lentivirus group Seqid270 GIGLVIMLVTMAIVAAAGAS *o***oo*oo***o*o***o Human immunodeficiency virus 2 Seqid271 GVMVLGFLGFLAMAGSAMGA ooo***o**oooo*oooooo Simian immunodeficiency virus Seqid272 GVFVLGFLGFLATAGSAMGA oooo**oo*o*oo**ooooo Simian immunodeficiency virus others Seqid273 GAIVLGLLGFLGLAGSAMG *ooooooo*o*ooo**ooo Ovine lentivirus Seqid274 GIGLVIVLAIMAIIAAAGAGLGVANAVQ Arteriviridae Porcint Reproductions og Respirations Syndrome (PRRS) PRRS Type I seqid275 SRKLGRSLIPHSCFWWLFLLC seqid276 GNGNSSTYQYIYNLTIC seqid277 GTAWLSTHFSWAVETFVLYHILSL seqid278 GFLTTSHFFDTLGLGAVSITGFC seqid279 RYAHTRFTNFIVDDRGRIHRW PRRS Type II seqid280 SNNNSSHIQLIYNLTLC seqid281 GTDWLAQKFDWAVETFVIFPVLTH seqid282 GALTTSHFLDTVGLATVSTAGYY seqid283 IYAVCALAALICFVIRLAKNC seqid284 VSTAGYYHGRYVLSSIYAVCALAALICFVIR L

[0260] All cited references are incorporated by reference.

[0261] The accompanying Figures and Examples are provided to explain rather than limit the present invention. It will be clear to the person skilled in the art that aspects, embodiments and claims of the present invention may be combined.

EXAMPLES

Example 1: ELISA

TNF-α ELISA assay

[0262] The supernatant from THP-1 cells treated with peptides was assayed on human TNF-α ELISA Max™ Deluxe Set (Biolegend, #430205). ELISA assay was performed according to the manufacturer’s protocol, as follows. Each incubation step was followed by sealing and shaking on the rotating table at 150-200 rpm, except the overnight incubation with the Capture Antibody, where plates were not shaken. One day prior running ELISA the 96-well assay plates were covered with the Capture Antibody, diluted 1:200 in 1x Coating Buffer (5x Coating Buffer diluted in ddH.sub.2O). 100 .Math.L of this Capture Antibody solution was added into all wells, sealed and incubated overnight (16-18 hrs) at 4° C. The next day all reagents from the set were brought to the room temperature (RT) before use. The plate was washed 4 times with minimum 300 .Math.L Wash Buffer (1x PBS, 0,05% Tween 20) per well. The residual buffer in the following washing was removed by blotting the plates against the absorbent paper. Next 200 .Math.L of the 1x Assay Diluent A (5x Assay Diluent A diluted in PBS pH = 7.4) was added for 1 h to block non-specific binding. While the plate was being blocked, all samples and standards (mandatory for each plate) were prepared. Standards and samples were run in triplicates. 1 mL of the top standard 250 pg/mL was prepared in 1x Assay Diluent A (1x AD) from the TNF-α stock solution (55 ng/ mL). The six two-fold serial dilutions of the 250 pg/mL top standard were performed, with the human TNF-α standard concentration: 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.2 pg/mL, 15.6 pg/mL, 7.8 pg/mL and 3.9 pg/mL, respectively. 1x AD serves as the zero standard (0 pg/mL). After blocking the plate, washing was performed and 100 .Math.L standards and samples were assayed in triplicates and incubated for 2 h in RT. Samples were not diluted, the whole supernatant from the THP-1 cells was assayed. After washing, 100 .Math.L of the Detection Antibody was applied to each well, diluted 1:200 in 1x AD, and incubated for 1 hour. Plate was washed and followed by 30 minutes incubation with 100 .Math.L of Avidin-HRP solution per well, diluted 1:1000 in 1x AD. The final washing was performed 5 times with at least 30 seconds interval between the washings, to decrease the background. Next 100 .Math.L of the freshly mixed TMB Substrate Solution (10 mL per plate, 5 mL of each from 2 substrates provided in the set) was applied and left in the dark for 15 min. It needs to be observed to prevent signal saturation, positive wells turned blue. After incubation in the dark the reaction was stopped with 100 .Math.L of 2N H.sub.2SO.sub.4 per well. Positive wells turned yellow. Absorbance was read at 450 nm and 570 nm (background) within 30 minutes. The data were analyzed in the Microsoft Excel 2010 program.

Example 2: Effect of Peptides on Cytokine and Transcription Factor mRNA Level Measurements by QPCT

Cell Culture

[0263] THP-1 cells were cultured in RPMI medium supplemented with 10% fetal bovine serum 2 mM glutamine, 100 U/ml penicillin, 100 .Math.g/ml streptomycin and used before passage 10. Cells were cultured in a humidified atmosphere in 95% air, 5% CO.sub.2 at 37° C.

RNA Isolation

[0264] RNAs from THP-1 cells were isolated using RNeasy ® Plus Mini Kit (Qiagen, DK) according to the manufacturer’s protocol. Quality and integrity of isolated RNA samples was controlled by determining A.sub.260/A.sub.280, A.sub.260/A.sub.230 absorbance ratios and 28S/18S rRNA ratios followed by rigorous DNase I (Ambion® TURBO DNA-free™) treatments.

Quantitative Real-Time RT-PCR

[0265] 500 ng total RNA was used for cDNA synthesis using iScript™ cDNA synthesis kit (Bio-Rad, CA USA) according to the instructions of the manufacturers. Real-time Q-PCR analysis was performed using a LightCycler 480 cycler (Roche Diagnostics, DK). 2 .Math.l of cDNA (from a total 20 .Math.l reaction volume) was used in a 20 .Math.l reaction. The real-time Q-PCR reactions contained 10 .Math.l SybrGreen 2x Master Mix (Roche Diagnostics, DK), 2 .Math.l forward primer (5 pmol/.Math.l), 2 .Math.l reverse primer (5 pmol/.Math.l) and 4 .Math.l water. After initial denaturation at 95° C. for 10 minutes, PCR amplifications were performed for 45 cycles. The primer sequences used in this study are shown in Table 1. The crossing point (CP) for each transcript was measured and defined at constant fluorescence level in Light Cycler 480 software. The mRNA levels for the test gene were normalized to the RPL13a or RPL37A value and relative quantification was determined using the ΔCt model presented by PE Applied Biosystems (Perkins Elmer, Foster City, CA USA). For quantitative real-time RT-PCR analysis, standard deviations were calculated and a T-test was employed to compare expression levels. P-values ≤ 0.05 were considered statistically significant.

TABLE-US-00003 Target gene/primer name Primer sequence 5′-3′ IL-2 β forward GTGGCAATGAGGATGACTTGTTC (SEQ ID NO:288) IL-2 β reverse TAGTGGTGGTCGGAGATTCGTA (SEQ ID NO:289) IL-6 forward AGCCACTCACCTCTTCAGAAC (SEQ ID NO:290) IL-6 reverse GCCTCTTTGCTGCTTTCACAC (SEQ ID NO:291) IL-10 forward GTGATGCCCCAAGCTGAGA (SEQ ID NO:292) IL-10 reverse CACGGCCTTGCTCTTGTTTT (SEQ ID NO:293) TNF-alpha forward CTGCTGCACTTTGGAGTGAT (SEQ ID NO:294) TNF-alpha reverse AGATGATCTGACTGCCTGGG (SEQ ID NO:295) NF-κB forward TGAGTCCTGCTCCTTCCA (SEQ ID NO:296) NF-κB reverse GCTTCGGTGTAGCCCATT (SEQ ID NO:297) RPL13a forward CATCGTGGCTAAACAGGTACTG (SEQ ID NO:298) RPL13a reverse GCACGACCTTGAGGGCAGCA (SEQ ID NO:299) RPL37A forward ATTGAAATCAGCCAGCACGC (SEQ ID NO:300) RPL37A reverse AGGAACCACAGTGCCAGATCC (SEQ ID NO:301)

Treatment of Cells/Induction of Cytokines

[0266] Pro-and anti-inflammatory cytokine gene expression was analyzed in un-differentiated THP-1 cells, designed as THP-1 monocytes. LPS is widely used as a potent and prototypical inducer of cytokine production in innate immunity which begins with the orchestration of monocytes. Pathogen associated molecular patterns (PAMPs), like lipopolysaccharide (LPS), play a pivotal role in initiation of variety of host responses caused by infection with Gram-negative bacteria. Such action leads to systemic inflammatory response, for instance up-regulation of pro-and anti- inflammatory cytokines, resulting in secretion of cytokine proteins into the blood stream.

[0267] THP-1 cells (1.0 x 10.sup.6) were cultured in a 24-well tissue culture plate (Corning). Cells were cultured with stimulant LPS at 1 .Math.g/ml with or without indicated peptides (at the indicated concentrations) for 4 h. LPS and peptides concentrations were chosen according to our preliminary optimization studies. RPMI 1640 medium containing 10% fetal bovine serum, 2 mM glutamine, 100 U/ml penicillin, 100 .Math.g/ml streptomycin was used as a control. To investigate gene expression and cytokine secretion cells were harvested at 4 h time point, while cell-free culture supernatants were collected and stored at -80° C. The time point of 4 h has been chosen based on the previously published gene expression and cytokine secretion kinetics of THP-1 monocytes stimulated with LPS.sup.1. The experiments were performed by two independent biological replications, started from a new batch of cells.

[0268] 1. Wasaporn Chanput, Jurriaan Mes, Robert A.M. Vreeburg, Huub F.J. Savelkoul and Harry J. Wichers. Transcriptional profiles of LPS-stimulated THP-1 monocytes and macrophages: a tool to study inflammation modulating effects of food-derived compounds. Food Funct., 2010, 1, 254-261.

Example 3

[0269] Inflammatory shock as a consequence of LPS release remains a serious clinical concern. In humans, inflammatory responses to LPS result in the release of cytokines and other cell mediators from monocytes and macrophages, which can cause fever, shock, organ failure and death. Here we present data that show that pretreatment of cells with INF F#2 results in a decrease in the release of cytokines including pro-inflammatory cytokines such as TNFalpha and IL-6. Therefore, treatment of patients, in the risk of developing sepsis, with INF F#2 could act beneficially to decrease production of proinflammatory cytokines and hereby lessen the risk of developing shock, organ failure and death. See FIG. 8.

[0270] The content of the ASCII text file of the sequence listing named “Sequence-Listing-as-filed-12397-0802”, having a size of 257 kb and a creation date of 13 Dec. 2022, and electronically submitted via EFS-Web on 13 Dec. 2022, is incorporated herein by reference in its entirety.

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