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COMPOSITIONS AND METHODS FOR THE PROPHYLAXIS AND TREATMENT OF BABESIOSIS

20220395564 · 2022-12-15

    Inventors

    Cpc classification

    International classification

    Abstract

    Described herein are compositions that comprise one or more Babesia microti antigens, one or more Babesia microti nucleic acid molecules, or one or more anti-Babesia microti antibodies and uses thereof in methods for the prophylaxis of babesiosis, the treatment of babesiosis and the monitoring of individuals undergoing prophylactic or therapeutic administration of the compositions of the invention.

    Claims

    1. A composition comprising (a) one or more Babesia microti (Bm) antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; and (b) a pharmaceutically acceptable carrier or diluent.

    2. The composition of claim 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    3. The composition of claim 2, further comprising one or more Bm antigens each comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    4. The composition of claim 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    5. The composition of claim 1, wherein the composition comprises two or more Bm antigens or antigenic fragments thereof.

    6. The composition of claim 5, wherein the composition comprises three or more Bm antigens or antigenic fragments thereof.

    7. The composition of claim 6, wherein the composition comprises four, five, six, seven, eight, or more Bm antigens or antigenic fragments thereof.

    8. The composition of claim 7, wherein the composition comprises nine or more Bm antigens or antigenic fragments thereof.

    9. The composition of claim 8, wherein the composition comprises twenty-four Bm antigens or antigenic fragments thereof.

    10. The composition of claim 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    11. The composition of claim 2, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    12. The composition of claim 3, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    13. The composition of claim 4, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    14. The composition of claim 1, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-24.

    15. The composition of claim 2, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-3.

    16. The composition of claim 3, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 4-9.

    17. The composition of claim 4, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-9.

    18. A composition comprising (a) one or more nucleic acid molecules encoding one or more Bm antigens, or one or more antigenic fragments thereof, and optionally (b) a pharmaceutically acceptable carrier or diluent, wherein the one or more Bm antigens, or the one or more antigenic fragments thereof, are as specified in claim 1.

    19. The composition of claim 18, wherein the one or more nucleic acid molecules comprise an RNA molecule or a DNA molecule, which optionally comprises a sequence of any one or more of SEQ ID NOs: 25-48, a fragment thereof, or a codon-optimized version thereof.

    20. The composition of claim 1, further comprising an adjuvant.

    21. The composition of claim 1, wherein the composition is formulated for administration by the oral route or a parenteral route, such as a parenteral route selected from the group consisting of the intravenous, intraperitoneal, subcutaneous, intramuscular, and topical routes.

    22. A composition comprising one or more antibodies that each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    23. The composition of claim 22, wherein the one or more antibodies each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    24. The composition of claim 23, further comprising one or more antibodies that each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    25. The composition of claim 22, wherein the one or more antibodies each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    26. The composition of claim 22, further comprising a pharmaceutically acceptable carrier or diluent.

    27. A method of immunizing or conferring protective immunity against babesiosis to a subject, the method comprising administering the composition of claim 1 to the subject.

    28. A method of immunizing or conferring protective immunity against babesiosis to a subject, the method comprising administering the composition of claim 22 to the subject.

    29. The method of claim 27, wherein the subject does not experience babesiosis.

    30. The method of claim 29, wherein treatment reduces the parasite burden upon a subsequent challenge.

    31. The method of claim 29, wherein treatment reduces the severity of babesiosis upon a subsequent challenge.

    32. The method of claim 28, wherein the subject experiences babesiosis.

    33. The method of claim 32, wherein treatment reduces the parasite burden.

    34. The method of claim 32, wherein treatment reduces the severity of babesiosis.

    35. The method of claim 27, wherein the subject experiences mild or severe babesiosis, including persistent or relapsing babesiosis.

    36. A method of supplementing an immune response in a subject experiencing babesiosis, the method comprising administering to the subject the composition of claim 22.

    37. A method of treating babesiosis in a subject in need thereof, the method comprising administering to the subject the composition of claim 22.

    38. The method of claim 27, wherein the etiology of babesiosis is Babesia microti.

    39. A method for determining whether a subject, prior to administration of a composition of claim 1, has protective immunity against Bm-induced babesiosis, the method comprising: a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; b. applying an antibody detection agent to the solid support of (a); and c. identifying the subject as likely to have protective immunity against Bm-induced babesiosis if the fluid sample from the subject is determined to have a sufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof of, that is, a titer in the fluid sample above a cutoff titer for the one or more Bm antigens.

    40. A method for determining whether a subject, following administration of a composition of claim 1, has acquired protective immunity against Bm-induced babesiosis, the method comprising: a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; b. applying an antibody detection agent to the solid support of (a); and c. identifying the subject as likely to have protective immunity against Bm-induced babesiosis if the fluid sample from the subject is determined to have a sufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof of, that is, a titer in the fluid sample above a cutoff titer for the one or more Bm antigens.

    41. A method for identifying a patient who experiences babesiosis and is likely to benefit from an anti-Bm antibody-based therapy, the method comprising: a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; b. applying an antibody detection agent to the solid support of (a); and c. identifying the patient as likely to benefit from an anti-Bm antibody based therapy if the fluid sample from the subject is determined to have an insufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof, that is, a titer in the fluid sample below a cutoff titer for the one or more Bm antigens.

    42. A method of optimizing the administration or therapeutic efficacy of an anti-Bm antibody-based therapy to a subject experiencing babesiosis, the method comprising: a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; b. applying an antibody detection agent to the solid support of (a); and c. administering to the subject the composition of claim 22 if the sample from the subject is determined to have an insufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof, that is, a titer below a cutoff titer for the one or more Bm antigens.

    43. The method of claim 41, further comprising administering to the subject the composition described herein, wherein the subject has been determined as likely to benefit from an anti-Bm antibody-based therapy or from a modified dose or dosage of anti-Bm antibody-based therapy.

    44. The method of claim 27, wherein the subject is a human.

    45. A kit comprising (a) one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof, wherein one or more Bm antigens are immobilized on one or more solid supports; (b) an antibody detection reagent; and (c) a package insert comprising instructions for using the one or more Bm antigens and the antibody detection reagent in accordance with the method of claim 40.

    46. A kit comprising (a) the composition of claim 1 and (b) a package insert comprising instructions for using the composition in accordance with a method described herein.

    47. A kit comprising (a) the composition of claim 22 and (b) a package insert comprising instructions for using the composition in accordance with a method described herein.

    Description

    BRIEF DESCRIPTION OF THE DRAWINGS

    [0084] FIG. 1 is a pair of graphs showing that B cells do not contribute to the resistance of wild-type mice to Babesia microti (B. microti) (left panel) but are critical for resolution of B. microti infection in cd4-deficient mice (right panel). Parasitemia values are reported as mean+SEM. N=5-7 per group.

    [0085] FIG. 2 is a series of graphs showing that resolution of B. microti infection in cd4-deficient mice is concomitant with an accumulation of B. microti-specific IgGs in blood. Parasitemia values and B. microti specific antibody titers are reported as mean+SEM. For a given time point, differences in antibody titers were tested for statistical significance using an unpaired Student's t-test (*=P<0.05; **=P<0.01; ***=P<0.001). N=6-7 per group.

    [0086] FIG. 3 is a graph showing that activation-induced cytidine deaminase, the enzyme required for antibody class switch and somatic hypermutation, is required for complete resolution of B. microti infection in cd4-deficient mice. Parasitemia values are reported as mean+/−SEM. N=7-8 per group.

    [0087] FIG. 4 is a graph showing that Fc receptors, whether activating or inhibitory, are dispensable for resolution of B. microti infection in cd4-deficient mice. Parasitemia values are reported as mean+SEM for 1 of the 4 groups; for clarity, SEM is omitted for 3 of the 4 groups. N=5-7 per group.

    [0088] FIG. 5 is a graph showing that the complement component C3 promotes clearance of B. microti organisms at time of peak infection in cd4-deficient mice but does not contribute to the resistance of wild-type mice to B. microti. Parasitemia values are reported as mean+/−SEM. For a given time point, differences in parasitemia values were tested for statistical significance using an unpaired Student's t-test (*=p<0.05; **=p<0.01; ***=P<0.001). N=6-8 per group.

    [0089] FIG. 6 is a graph showing the time points at which blood was collected from wild-type mice and cd4-deficient mice infected with B. microti. Specifically, blood samples were obtained at times (arrows) of peak infection (day 18), partial resolution of infection (day 24), and full resolution of infection (day 30) in cd4-deficient mice. Note that on day 24 post-infection, the infection was fully resolved in WT mice. N=6 for the WT group; N=10 for the cd4−/− group.

    [0090] FIG. 7A is a heat map of IgG antibody reactivity stratified by mouse strain and time post-infection. Each column of the heat map represents a mouse sample, and each row represents a B. microti protein. Each grey header encompasses several columns and denotes a mouse strain at a given time point. B. microti proteins (n=74) are those significantly reactive to IgG contained in plasma from B. microti-infected wild-type or cd4-deficient mice and are ranked in decreasing order by mean normalized signal intensity for all samples across strains and time points. The grey scale (top-left corner) represents normalized signal intensity on the log 2 scale, i.e., values of 1.0 signify a 2.0-fold increase over background, values of 2.0 signify a 4.0-fold increase over background, values of 3.0 signify an 8.0-fold increase over background, etc.

    [0091] FIG. 7B is a Venn diagram showing the numbers of IgG reactive B. microti proteins at times of peak infection (inner circle, n=16), partial resolution of infection (intermediate circle, n=35), and full resolution of infection (outer circle, n=52) in cd4-deficient mice.

    [0092] FIG. 8 is a series of graphs showing the IgG repertoire against B. microti antigens in wild-type mice (top panel) and cd4-deficient mice (middle panel). Dashed lines indicate the cutoff for IgG reactivity (2-fold above background). The lower panel depicts strain differences in IgG reactivity. In this panel, dashed lines depict strain differences (gain or loss) in IgG reactivity of 2.0-fold.

    [0093] FIG. 9 is a pair of graphs showing the IgG reactivity against B. microti antigens in cd4-deficient mice at time of full resolution of infection (top panel) and time of peak infection (bottom panel). Dashed lines indicate the cutoff for IgG reactivity (2-fold above background). In both panels, B. microti antigens are ranked in decreasing order of IgG reactivity at time of full resolution. Solid grey bars in the bottom panel denote antigens for which the IgG reactivity is above the cutoff defined above. Below each grey bar is reported the rank of the antigen in regard to IgG reactivity at time of full resolution.

    [0094] FIG. 10 is a table that lists B. microti antigens for which IgG reactivity is detected at time of peak infection in cd4-deficient mice. Antigens are considered immunoreactive when IgG binding is above that of background by at least 2-fold (1.0 on a log 2 scale). Antigens are identified by their gene ID and are ranked in decreasing order of log 2 IgG reactivity. A brief description of each antigen, when available, is provided. This description includes the Plasmodium falciparum ortholog in the clone PF3D7, when available. The rank of each antigen in regard to IgG reactivity at time of full resolution is reported in the most right column.

    [0095] FIG. 11 is a series of graphs showing and comparing the IgG reactivity against B. microti antigens in cd4-deficient mice at times of peak infection (top panel) and partial resolution of infection (middle panel). In the top and middle panels, dashed lines indicate the cutoff for IgG reactivity (2-fold above background). The lower panel depicts differences (e.g., gains and losses) in IgG reactivity from time of peak infection to time of partial resolution. In this panel, the dashed line depicts gains in IgG reactivity of 2-fold. Grey bars denote antigens that gained reactivity from time of peak infection to time of partial resolution. Solid grey bars denote antigens that were already reactive at time of peak infection, whereas hatched grey bars denote antigens that were not reactive at time of peak infection. Below each grey bar is reported the rank of the antigen in regard to IgG reactivity at time of full resolution.

    [0096] FIG. 12 is a volcano plot showing gains in log 2 IgG reactivity (from time of peak infection to time of partial resolution) against −log 10 p values. The larger dots depict the 20 antigens for which gains in IgG reactivity reached the highest degree of significance as assessed by use of a paired Student's t-test. The horizontal dashed line indicates the threshold for significance (p=0.05). Numbers denote antigens for which the gain in reactivity was significant and greater than 2-fold (vertical dashed line). Numbers highlighted by a circle denote antigens that were already reactive at time of peak infection, whereas numbers highlighted by a hexagon denote antigens that were not reactive at time of peak infection. Each number corresponds to the rank of the antigen in regard to IgG reactivity at time of full resolution.

    [0097] FIG. 13 is a table that lists B. microti antigens for which IgG reactivity increased by more than 2-fold from time of peak infection to time of partial resolution in cd4-deficient mice. Antigens are identified by their gene ID and ranked in decreasing order of the gain in log 2 IgG reactivity. A brief description of each antigen, when available, is provided. This description includes the Plasmodium falciparum ortholog in the clone PF3D7, when available. The rank of each antigen in regard to IgG reactivity at time of full resolution is reported in the most right column. Antigens denoted in grey were already reactive at time of peak infection, whereas antigens denoted in black were not reactive at time of peak infection.

    [0098] FIG. 14 is a series of graphs showing and comparing the IgG reactivity against B. microti antigens in cd4-deficient mice at times of partial infection (top panel) and full resolution of infection (middle panel). In the top and middle panels, dashed lines indicate the cutoff for IgG reactivity (2-fold above background). The lower panel depicts differences (e.g., gains and losses) in IgG reactivity from time of partial resolution to time of full resolution. In this panel, the dashed line depicts gains in IgG reactivity of 2-fold. Grey columns denote antigens that gained reactivity from time of partial resolution to time of full resolution. Solid grey columns denote antigens that were already reactive at time of peak infection, whereas hatched grey columns denote antigens that were not reactive at time of peak infection but gained reactivity from time of peak infection to time of partial resolution. The dotted grey column denotes the single antigen that was not reactive at time of peak infection, did not gain reactivity from time of peak infection to time of partial resolution but gained reactivity from time of partial resolution to time of full resolution. Below each grey bar is reported the rank of the antigen in regard to IgG reactivity at time of full resolution.

    [0099] FIG. 15 is a volcano plot showing gains in log 2 IgG reactivity (from time of partial resolution to time of full resolution) against −log 10 p values. The larger dots depict the 20 antigens for which gains in IgG reactivity reached the highest degree of significance as assessed by use of a paired Student's t-test. The horizontal dashed line indicates the threshold for significance (p=0.05). Numbers denote antigens for which the gain in reactivity was significant and greater than 2-fold (vertical dashed line). Numbers highlighted by a circle denote antigens that were already reactive at time of peak infection, whereas numbers highlighted by a hexagon denote antigens that were not reactive at time of peak infection but gained reactivity from time of peak infection to time of partial resolution. The number highlighted by a rectangle denotes the single antigen that was not reactive at time of peak infection, did not gain reactivity from time of peak infection to time of partial resolution but gained reactivity from time of partial resolution to time of full resolution. Each number corresponds to the rank of the antigen in regard to IgG reactivity at time of full resolution.

    [0100] FIG. 16 is a table that lists B. microti antigens for which IgG reactivity increased by more than 2-fold from time of partial resolution to time of full resolution in cd4-deficient mice. Antigens are identified by their gene ID and ranked in decreasing order of the gain in log 2 IgG reactivity. A brief description of each antigen, when available, is provided. This description includes the Plasmodium falciparum ortholog in the clone PF3D7, when available. The rank of each antigen in regard to IgG reactivity at time of full resolution is reported in the most right column. Antigens denoted in grey were reactive at time of peak infection and/or gained reactivity from time of peak infection to time of partial resolution. The antigen denoted in black was not reactive at time of peak infection, did not gain reactivity from time of peak infection to time of partial resolution but gained reactivity from time of partial resolution to time of full resolution.

    [0101] FIG. 17 is a series of graphs showing the inverse relationship between parasitemia and IgG reactivity against each of two B. microti antigens at time of peak infection in cd4-deficient mice. These 2 antigens, namely BMR1_01 G03280 and BMR1_01 G00985, ranked first and tenth when considering their IgG reactivity at time of full resolution in cd4-deficient mice (top panel).

    [0102] FIG. 18 is a series of graphs showing the relationship between parasitemia and IgG reactivity against each of four B. microti antigens at time of partial resolution in cd4-deficient mice. Three antigens, namely BMR1_01 G03280, BMR1_04 G05532, and BMR1_04 G07360, had IgG titers at time of partial resolution which were inversely correlated with parasitemia at time of partial resolution (left bottom panels). A fourth antigen, namely BMR1_01 G02100, had IgG titers at time of partial resolution which were positively correlated with parasitemia at time of partial resolution (most right bottom panel). Note that BMR1_01 G03280 and BMR1_04 G07360 were already reactive at time of peak infection (solid grey) whereas BMR1_04 G05532 and BMR1_01 G02100 were not reactive at time of peak infection but gained reactivity from time peak of infection to time of partial resolution (hatched grey).

    [0103] FIG. 19 is a schematic that classifies the 24 B. microti antigens which displayed IgG reactivity during the resolution of B. microti infection in cd4-deficient mice. Sixteen antigens were identified as IgG reactive at time of peak infection. Of the antigens identified as gaining IgG reactivity from time of peak infection to time of partial resolution, only 7 were not reactive at time of peak infection. Of the antigens identified as gaining IgG reactivity from time of partial resolution to time of full resolution, only 1 antigen was not reactive at time of peak infection or had not gained reactivity by time of partial resolution. Among these 24 distinct antigens, 15 are predicted to lack a signal peptide and are referred to as “group #3 antigens.” Of the 9 antigens predicted to contain a signal peptide, only 3 had an IgG reactivity which was inversely correlated with parasitemia at time of peak infection or time of partial resolution. These 3 antigens are referred to as “group #1 antigens.” The other 6 antigens are referred to as group #2 antigens as they are predicted to contain a signal peptide but displayed IgG reactivity at time of peak infection or partial resolution which was not inversely correlated with parasitemia measured at the respective time point.

    [0104] FIG. 20 is a table that lists the 24 B. microti antigens identified as displaying or gaining IgG reactivity during the resolution of B. microti infection in cd4-deficient mice. Antigens are identified by their gene ID. A brief description of each antigen, when available, is provided. This description includes the Plasmodium falciparum ortholog in the clone PF3D7, when available. Antigens are classified into three groups using two criteria, namely the predicted presence of a signal peptide, and an inverse relationship between IgG reactivity and parasitemia at time of peak infection and/or partial resolution of infection. Within each group, antigens are ranked in decreasing order of their highest IgG reactivity or gain in IgG reactivity (grey boxes). For the purpose of this application, a sequence ID number (SEQ ID NO) is attributed to each of the 24 antigens.

    DETAILED DESCRIPTION OF THE INVENTION

    [0105] The invention is based, in part, on the findings that a) resolution of B. microti infection in cd4-deficient mice requires antibody-producing B cells and antibody class switching, b) such resolution is concomitant with the accumulation of B. microti-specific IgG antibodies in blood, and c) these IgG antibodies target a restricted set of B. microti polypeptides. Accordingly, the invention provides compositions for use in methods for the prophylaxis of babesiosis, the monitoring of prophylaxis efficacy, the treatment of babesiosis, and the monitoring of treatment regimens in a subject (e.g., a mammalian subject, such as a human), as well as such methods. The invention also provides kits that can be used to carry out the methods of the invention. The compositions, methods, and kits of the invention are described further, as follows.

    Compositions

    [0106] In general, the present invention provides compositions comprising one or more B. microti (Bm) antigens for use in methods for the prophylaxis of babesiosis, the monitoring of prophylaxis efficacy, and the monitoring of treatment regimens for babesiosis that comprise one or more anti-Bm antibodies. The present invention also provides compositions comprising one or more nucleic acid molecules (e.g., RNA or DNA) encoding one or more Bm antigens for use in methods for the prophylaxis of babesiosis. The present invention further provides compositions comprising one or more anti-Bm antibodies for use in methods for the prophylaxis and the treatment of babesiosis.

    [0107] These compositions and methods can be used to generate or provide immunity to B. microti and thereby to confer partial, enhanced, or full protection in humans and other mammalian subjects who are at risk of exposure to Bm and at risk of developing babesiosis (e.g., mild or severe babesiosis, including persistent or relapsing babesiosis). In various examples, the compositions are useful to (a) reduce the chance of a subject to become ill when infected with Bm, (b) reduce the severity and/or duration of illness in an infected subject, (c) reduce the parasite burden in an infected subject, and/or (d) reduce the chance of dying from babesiosis. In various examples, the compositions are vaccines.

    [0108] Babesia microti Antigen-Based Compositions

    [0109] Compositions of the invention, which can be used, e.g., in the methods and kits described herein, include one or more Bm antigens and optionally a pharmaceutically acceptable adjuvant, carrier or diluent.

    [0110] Exemplary Bm antigens for use in the compositions of the invention include BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_01 G00985 (SEQ ID NO: 3), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7), BMR1_04 G06070 (SEQ ID NO: 8), BMR1_04 G09385 (SEQ ID NO: 9), BMR1_03 G04485 (SEQ ID NO: 10), BMR1_03 G01645 (SEQ ID NO: 11), BMR1_03 G01960 (SEQ ID NO: 12), BMR1_04 G05080 (SEQ ID NO: 13), BMR1_03 G00820 (SEQ ID NO: 14), BMR1_01 G02100 (SEQ ID NO: 15), BMR1_04 G07360 (SEQ ID NO: 16), BMR1_02 G02185 (SEQ ID NO: 17), BMR1_04 G08260 (SEQ ID NO: 18), BMR1_03 G02345 (SEQ ID NO: 19), BMR1_02 G02960 (SEQ ID NO: 20), BMR1_01 G02545 (SEQ ID NO: 21), BMR1_03 G04110 (SEQ ID NO: 22), BMR1_02 G02560 (SEQ ID NO: 23), and BMR1_04 G05635 (SEQ ID NO: 24). The amino acid sequences for the 24 full-length antigens or their ectodomains are provided in Table 1.

    TABLE-US-00001 TABLE 1 Amino acid sequences of exemplary full-length B. microti antigens or their ectodomain SEQ ID NO. Amino Acid Sequence Gene ID 1 DVYEISSGNPPDIEPTSTSLETNVVTNYIPEPNADSESVHVEIQEHDNINPQDAC BMR1_01G03280 DSEPLEQMDSDTRVLPESLDEGVPHQFSRLGHHSDMASDINDEEPSFKIGENDII ectodomain QPPWEDTAPYHSIDDEELDNLMRLTAQETSDDHEEGNGKLNTNKSEKTERKSH DTQTPQEIYEELDNLLRLTAQEIYEERKEGHGKPNTNKSEKAERKSHDTQTTQEI CEECEEGHDKINKNKSGNAGIKSYDTQTPQETSDAHEEGHDEINTNKSEKAERK SHDTQTTQEICEECEEGHDKINKNKSGNAGIKSYDTQTPQETSDAHEEEHGNLN KNKSGKAGIKSHNTQTPLKKKDFCKEGCHGCNNKPEDNERDPSSPDDDGGCEC GMTNHFVFDYKTTLLLKSLKTET 2 IITDGLSAIGSVASEVGNTVKDVSSEALLGELQQIADGGKIIRDGTESNFVNSIANT BMR1_04G05532 VMKNVVGTAVLKASSGITHNGDFAFYNFMYPENDDYPWACICDESDYEEYIKG ectodomain KKDKVRCRNYIDSSLQNAVLYCNPANHNSSINDNANNNPPKQIDNHVSIPQTA PANHTTVLSTEVDTNHNEQKQPNSPSVPSESQNSVSAPKDESVSSTVEGAKSSS 3 TLVSKITTPNNLPGIDTCSNWEDVSVCTTKNTRNCISGEGKPKDCFAIGKSLFKTF BMR1_01G00985 PNCYEGVIIDQVTFSGFETLEIHYCDIDKILHKANEVVKSIHELKEKTNQLTEKIKDI ectodomain PDLIDKVIKVNSEISKIFHQDKIKHMEKEANDFKNALKTTRNYIISYDSLDKTKQSN LLTSLGKLMNKIKTKLSEMDKTLHSTLDTNNTIIDLVNNNSSHVKHPNDFNKTM ELYNETITKADAIKKNIEKLKEHRKISTHKTIFSNNIDKLIDNLTDYFENINRSIDAVR DKLSKYQLETGKMVLLFKNVNEIQKHIKNTDMHIRTCNYDFSDIEQKYSLITAKIT VEDGQSITTSNKSTVDIPEEKVDRVNVNVDKAENSDNETSQENTSVKPTDHKEI EDSASEENAIGENGDYDSDEDIDTNDVKEDHENAIDSEYTVSSTGDVLEDEVVE ENAIGENGDYDSDEDIDTNDVKEDHENAIDSEYTVSSTGDVLEDEVVEENAIGE NGDYDSDEDIDTNDVKEDHENAIDSEYTVSSTGDVLEDEVVEENAIGENGDYDS DEDIDTNDVKEDHENAIDSEYTVSSTGDVLEDEVVEENAIGENGDYDSDEDIDT NDVKEDHENAIDSEYLVSSTGDVLEDEVVEENAIGENGDYDSDEDIDTNDVKED HEDAIDSEYLVSLTGDVLEDEVVEENAIGENGDHDSDEDIDIEEVNEEDHEDAID SDNSISNSENVDTTPTENVDTIPTKNANTTPTKNANATPTKNVDTIPTKNVDTIP TKNANTTPTKNVVTTPTKNANTTPTKNVVTTPTKNANTTPTKNVVTTPTKNAN TTPTKNVVTTSTGNVRTKHTSTNSHVLAPDTDEYPAQISQHKTIDKYYQSLELED EENADISSSDKPVSPINLEDKNSTYDHMHKTDNVKSAGIASYD 4 NRSCPGNNGVGGGSGDNNSGIIPNDPHPCCNNLRQKPQYQTKPENELVNDDR BMR1_02G01760 DLNFNKIRGGKQIITFTVPSIDDLKNKRLSDSEFILSEKANPLISSGDSKNVIVFEVK ectodomain NDNEKLMGSVEVGQWEVTITTSCIRRMFDSNEVSDNIPMYIYIVDYFEGGNST VSKFFFANNRWNDFTNHTPNAA 5 FLLNRSEFKWFKVGLIITTIFPFKHSFDYNLTHIFLFSICTLIFCVKPVDEESGAKKEG BMR1_03G00365 FDFKKMVPDKFKKYT ectodomain 6 DSGNSSPQTPPETSSPINGVIGDENNGLEHLSSSGLEVDDNLPELLKTSPFSGQN BMR1_03G04695 SDVQSASTPVEPTTPVHSNDQSNPITNKVDTNSNDHTDIKNEGSSHRTSSNNSS ectodomain VTTNTNNEIRNGGGPLDQNEDKAEDEGETDAEGRGWNERTKNKPTFNATNR DFVDDNLPELLKTSPFSGQNSDVQSASTPVEPTTPVHSNDQSNPITNKVDTNSN DHTDIKNEGSSHRTSSNNSSVTTNTNNEIRNGGGPLDQNEDKAEDEGETDAEG RGWNERTKNKPTFNATNRASPDGIGKMNMEEKQLENFINVSSNALELDISIGR DNFATKFLAQHVNIFGDRISGLSAAYVEGYNNLAKIMYNSHSVLFDRKFNGAVIS DNLIGNIADFGSYFLEISPNTTRTNRSDYLKSVVLSKVQYLLSADFSTTDNIQRLTN LALALGYNNVKENNPGNSQHSITTSLSTELFWSFGNNIFLFGHLATLMLAYLESN AYFTSGATRPFFSWQTLVSTGGNEKFDKLDSMCGVIRGSKYSRKNNGFIKPHYK RLRRKTLLEGEPRLLCSMLEEALDTVDKAIKFKGEELNSQGANIENSVSNDINSKR LQAKLCSNLNDSLINVSCDFRSSKLDKHNKKLREAFDLLLACGNLNTGKKEAFPE YLRLISNPFEYGDIFSMTMWWDPREFDGKQGWVEIYKKLRKNIMKPELKNVD MQLKYDSAISYYKQLKESETYPKKNIPWARLYLYMSVIMSRSNAMSWAEDALR SFSNLYRMKPSLVMRGEGLETLLNYCAPDPVALSHIFLYHFLTKKDAGKDLEKDL RRLEKGTLLSRIVNSSSIFIPNKLKKFLKMGARGFFNKKLNTLRAKSTLLRLFPKNLL HSALGAIEFTTHSLATLQISKNMDMWESLAQTKNLDAGGFPGEIDSLFNHWSE SGGYSGYITGKLENGDDLTGDDIKKMNIKAPINNDSLNWQKYINKKISEHFGKFL NLPFIQASGSQKNYIYQLVRDSKANLDDNLEQTVFFGKVLPPGKTNNVIKKLKRI ADSFTSMLLRSSARPVDHAVWVGVKINVPIVIHITKKLYMIQRDMPRKEAWNL ESAFLDLLQDLVIMVTNPGKRSPIGFETIGGNPGLPEISIRYPHMSIEERKIEFQHS QCADHCISIWRSLIAFTLNTLNNPAAIKQFEKSLSSNSSLNDMSKPEYINSFKYILK GDSVLHMYDNMLPRKVKREIKALKYGK 7 LPDILSQNNTFKSFLEVNNVDQEDLICNKALCKSTDSINRNTSSYCYKYKLCSKCS BMR1_03G03430 VSNVPDHPVCYLLDNDHNYIHLMEGHLGSQPIGSANSHDNSSHDEHSSHSNN ectodomain GDMMDEHEEENFLQEYESKSMKFIPTSNMSDFDHARRSCAVDSKGNVMISVR LIIQWYMSKDKSNNQQHHGNDDDSQNYDANYLQLTPMYSDDSVNSSMLEM DHDDSESSNSHKSRMANMAKNFQVLKNIHKSAVKRYKSPKAKIYLIFSNPKINSC RHPVIYNGKISPSSMFVAKLESTISQIDLTQDLIKSSIETIVSCEACDKLKYNSCIQVT CAKNTPGAASLAMGSAVYVPMTNTTIGVNAHNPNAVVAAGIPMGKIPVIPHP AAISGGNVGHLNNGLHKAVNNAVMMPNGTSLPVQSGVVIKSLYNCLAFLLTIL YLNF 8 NPIFASATSAPSRNRAQDMTKAECIELLKGIVKSQEETKIVMKKLTSDLINNPLRL BMR1_04G06070 EQVYTKASQMQPEDPMEQWGVTVIDLDHLIEKYQHDPIVKDYILKIMNSPGIN ectodomain DTTLSDDVRNITISQILAIHEYMLSELESVVREFKSLQNRQTMEIKTLTIAAQAIVA AKVEEKFNLTSDQVESAVIINHAELTASHAFTRLTMOMQ TEMSELIGCQFPGW 9 STNGKGGVDSVSKKSFVIELEDSTFERKTQASSGGTSGVWFVKFYAPWCGHCR BMR1_04G09385 SMENDWNELANILGKQINVAKIDATKHSVTAKRFGITSFPTLLLLKDGNFYQYEN ectodomain NNRTADALKQFALHGYKQVKSKPVPKEWSYFVRFKFFIKSGFYEVKRIYQLAYPG FIT 10 MAKLHESTSSASASFDPEKSDYDDTYVLTETTPTYIRHGFVRKVFAILFAQLLVTL BMR1_03G04485 GFSLICYFYRESVHSFISKNIWIFPTLAILSFITSLILIFSPSLSRRYPLNYAILVIETLYFS full sequence FIVGLSCAFTKSPTAIVLSVSITLGIILLVVLFTLQTKIDFTRYIIYFILFSFVTLVFGFIGI FVPFDTPLRMFYYGLGVLGYSLWMVLDLQLIIGGKTYEWTVDDYVPASLSLYTD VIGIFLNVHGMFSDR 11 MTITLNIKVNSETNFTVEAEPSFTVKELKILCESQSNIEAQNQRLICKGKLLKDTDIL BMR1_03G01645 SDVGAVDGATVYLVRSQVNKTQSAAPKQNTVPQPTLQTTNQPAGQTQTSGLG full sequence FQQQGFQQGFSGSQQPGFQANPFQSMLAGGFPNLDPTQMMEILNSPMAQE AMQRLSQNPEVLRNILQNSSLMTPMLEQNPMLSEMLSNPELMRSMLRPEVL QAGLQMHQAMQQQQQQQPGTQTNPIGSQNPDFSNMMRQMMNVFQQN PSVAQPTAPQVYTDPRPPAERFATQLQALAEMGFIDTEKNITALIATNGDLNAT VTRLLESNF 12 MEEAERKFKEFNWADSQGWRIYWDNLYPTPPLSKVDKFKRSWFKRNVDSNIS BMR1_03G01960 STPLSEVGKQTQQTTPNQYRSQGNFAILTLEAGVRLLYLLITAPLFVLPLIGVRLSR full sequence YIYYHYYIDIALLLIFLLSGIVRERGLPKMDSVWLASAFYSDMTQYIMYTCILMMS APRPVYLILPFLTCLIGLNSLAETNLSKLPKWLENIVGEIFKYTKDNIYWLMQTRG DVECYLLFYIVFGFLTKSSAVITLMAYINFMKLRIGIGDPFIMSAFSKLHGCIVKLLS YDMFGHIPLRMYLKISELLTKYMSGPRPQY 13 MEDQNTTNESISNLHMENYFPKDIFSNLDNQKNLKTYHSKTFEKYFESAAKDNV BMR1_04G05080 ERCRTSAVKEWLNRNLPSEYNERHKPTLKLNLSPNHLNSTYNKQISDYTRNAYN full sequence RIEQLKKLQEAYSLLRQKLQEKRRASCHKEVSEAEASVLNTQRLIISLKREDKDKIA TELLIQNAEELGIPSKDLETLNGYTFNEALKTIIDIISNMINNKFMTNLKDRCESAR RKAGSEYRDEMERIKVDLDMYKTKSEKLESTISTLNSYADSSKQNAEKVIVIELQH SLEDSNRQINQLKDSLYNMAKVNEELEKNEVKLEESLVELERYKLESQKLESVIKD LELLNQQKHDNEELIKMLHDELDQCKSKSLQLNKKIEDLENSNKENLIPLNNELA QAYQKLSELEHSYEQIEHQKNEADKKLESSIDEIEKQKQHSEELEQSITKLKQTIEE MEKETTDQVNDLQTCLIDANCKIESLEGTISQLKQLNLELEGGEHRIQELQSKLTE SNSTIEKLEKTITELENVSSKYIDYDEKMDNLQKQLKEYTEKITVLENNASEFNYKD QAETLKTELDKSQLKIMELETMIKENSEKTERVPSPRKDNEEVDRLTSEILQLKNQ VDILQNEKTDLEQKLSQQSPRKDNEEVDRLTSEILQLKNQVDILQNEKTDLEQKL SQQSPRKDNEEVDRLTSEILQLKNQVDILQNEKTDLEQKLSQQSPRKDNEEVDR LTSEILQLKNQVDILQNEKTDLEQKLSQQSPRKDNEEVDRLTSEILQLKNQVDILQ NEKTDLEQKLSQISAVIEENRQYKEKIELLERKLNEINKEKPKDSFTNVEVINAAFE DVRSLPINSSNASDWTAMPSELELEEHKLYKKKSSRKGKKKSSREKETSRSDISSR STSRHSKKDKPTIVDNNSTDVEASDSNEQMISDPVESITDQLNLVKQSTIQLTEL GYDSISNVGSYLSDYIFGGN 14 MDDLPGSLHQSTPNEKPMAPPTAPKSAPHVPISVESKELSKEIPKESPMTKEDKK BMR1_03G00820 DVAKSKVSAAVTEKKVVKEPVVPKITIEMKKFSMSRESTQYSIFIIFNLIFALLYIFK full sequence VRLMAIFCNLIIVAISIGAILSSIDPNRRKVDETNVNIIFISPTTVSDLAIVITAKLNQY ISYFRRILLWQDFILSTRFTLCVYIMGILFKIIPLVALIYIMCWAFYLYMFICKEMAD QLLDIIMVYLKRVSGNFDEFCCNIPKMKDVGKEL 15 MFNENEIPQFQRPFTPSKDTEIDTSINFPANSILNNISFQKLLDWLDECTEGLPIV BMR1_01G02100 ESFARRFNVTRGHIAAIFSAILILYFIFGWKINIFCNTIGLVYPAFKSHKVLLIHRAM full sequence TESPKATATITTGGKDKENDTNPQMPTCLNGIQGEIMFWLRYWIVYSLYLFISILI FPLISWLPLISIVRVGFILYLYHPYTRGANAIYYLVISPLLSKNQKIIEQAIDTLERVA MGEIRKFTEKQMKLH 16 MDSIEECNKLVDAVTKLATSFDYQAQEYLYNLTRDENAINIALSFLLSNKYVLNYS BMR1_04G07360 SSYSNEILQYITDVRGPSSCCIWDNIQLDENTSRHLGHITSILFKEHMHIVLCFDD full sequence AKLIALIDTLVTIWQLQSTISLVTQSCFADNATDDHITRVISMIIFKKPQLLTHFINIL NTPEVTDKYVADFSMKFKLRNCDRDDIYVTRYRHMIVYVLAQVLENFVSSNVTD KISFLPINFSDLVILSGNCNNRYLYSCALLIVNSLNFHISEELLHQLLVIVEKSGFDCD ILEMWNFAISHLQYLKVDNFMMKSFRLVQNGLIESTNPINIVPVLNGITVYYVN NSSVPTEIVRAYRQLLSVESLDIPIEIGELFAAVRNACTESLITYDKIASFVSQFDIR MISSRLIELCNPNMQCWSWLNVNCCVEEFPTDYDIYIDSMKNVFKDLFFILPEQ LLDTIISEILQSVNENTETSLIALICYDCLEVRHALAILDKIKSLFTLKLSSNYDFILTC MFCQSIFVPHLHRIKDILFQRFVINIVIKISNIWSKRLAKCIAILNDRDSVHSVIECV QYILETSKSKGVLSQNHYPSILSLSNLLDTLPQMPCDVNLTGSNYIFCKCLLALSIST RQLSQILPKIALNLDILYEGDVDNDLFCLSTSDSQAAVNLLMQLYIQNKVTDIPNE LHEIRHPGLLMLCNLNENFEQFRMNKCVEFMSTVSQESCCHCAWACIAYASYV IEQKFNFEFSMEMVKVIYRVLPCLIKMVKSKNEPLHWDLDAILEYRINEIHPSIKCL KGTIWNIYYDGTVSVGSDACKYLSLVNKYAPRIIKSLSNPNVLQLVYQISMCTGSI EVIANKIAISL 17 MSCILKCNNEDELVVDGEKPKVVEYVEKPSVIYKPTTVVPPNSLIEITAPKDLPQN BMR1_02G02185 PTFFPTIDTFFDNDVKQIVLLMELPGFVAGDIDLEVGEGEVCVCGPRSKEELYEKY full sequence GQNLDIHIRERKVGYFYRRFKLPHNALDNTVKASYQNGILEVRITCTEFSPKTRVE ITS 18 MNGSVEELLNRLSAINDRCYLLVDEIKNYMSNKLYHELTLALIELFTMSEISCNDR BMR1_04G08260 LLLFEMIVHPIKNDLNILKFSHILRLSSEHLEPLASLDQLSKYDNYLSTDTQASFIKIA full sequence KSYHHTRNQSYDQSLKLLEEVKPEIESGFGLDITVISAYYKVSANLNKATHKYNS WYQDSLMYLNYTPLDSISPTERDELALDIAIASIAAPDNYNFGAVLIQPLINTCLK QHSTFGWVYAILMALNDGDFTQYDEIISKYKVQISHSELNHHKEQLQRKITLMA FLKLVFRKAKKQRIFTFEEISQNCRIPIDEVEYLLLKAMCNNWKGKINQVEQIVSF TWVQPRIIDSTKLTVLLDGVNEWNQQLKALINKLKEITPELLVS 19 MSGLFGQSQQIGGGLFGQSNQQSGGGLFGSTSQQPTQTCSGLFGSSPAPANS BMR1_03G02345 SIFGSNTQSAASSGGIFGSSTAPVNSGGGIFGQSNTNVSSGSGLFGGGNTTGQS full sequence GGGIFGSSTTSAPASGGGLFGQTGTTTSGGGGLFTSSFAPAPSSGGLFGQPSTP ATSGTGLFSSTSTTOPSSGAGLFGSSTTPASGSGGLFGQPSTSTTTSGGIFGSSTT SAPASGGGLFGQTGTPASGSSGIFGSTNTTTSASGTGLFGSTSTTPQPGSGSGLF GGGNTTGQSGGGIFGSSTTSAPASGGGLFGQTGTTTSGGNLFGTTSTTTPAPAS GGLFGSSSTTSTTTPTQATTTVGGGGLFGTASATTAPASGGLFGTTSTTTPAPAS GGLFGSSSTTSTTPTQATTTVGGGGLFGTASATTAPVSGGLFGTTSTTTPAPAN NTTPANTTTVPTAILTTSTPSPATDGLFGSTDVTTTTDSTTKLVGTSPFEKQTDSG PDVTAATNDTPNAFITDKPTAGGDLKGQVELSFSSVEHECVQDLLSNWEKRME VKIQRFTEFAQDIQRIDRDLILQTEKLQVLLDEQATVQERQNQVQEMIQVIEKE QQQVLESLDIMDTALETLLGPDKKITSKSGETIDFVSNKLRDLEAQLKAAHDVVD SVVKASQPEPLANVAKVFAFHQDTIENIQLQTSEIEKKLDAIKQQQAV 20 MASLLKVSPQDNIEFPLVLYTPLNANLLLENLSGVHVAFKIKTTAPKGYLVRPSTG BMR1_02G02960 TIKPGEALTVQIILQPLSEVPNVVNDRFLVQCTAIANDELVSKDFWTTLDKASIQD full sequence HRLNVTFKKDIGLNIQTSQSNIGVPPHIAARILTPLGPNAGVAELRQKYEELVSYC LTAEKQKAALVKDNEKLRQRLHLGPNDPASGNKWPLEGWHLPVMVIILVIILKA IGYW 21 MSQGPAIGIDLGTTYSCVGVWKNETVEIIANDQGNRTTPSYVAFTDVERLVGD BMR1_01G02545 AAKNQDARNPENTVFDAKRLIGRKINDPCIQSDIKHWPFTVAAGPNDKPVIKV full sequence QFQGETKSFHPEEISSMVLTKMKEIAESYLGKTISNAVITVPAYFNDSQRQATKD AGTIAGLNVMRIINEPTAAAIAYGMDKKGTSEKNVLIFDLGGGTFDVSILTIEDGI FEVKATQGDTHLGGEDFDNRLVNFCVDDFKRKNGGKNISTNRRALRRLRTQCE RAKRTLSHSTQATIVVEAIFDGIDYSCNITRARFEELCAEMFKNTLIPVEKALADA DMDKKQIHEVVLVGGSTRIPKIQQLIKDFFNGKEPCKSINPDEAVAYGAAVQAAI LTGEQSSKVQDLLLLDVTPLSLGLETAGGVMTVLIPRNTTIPAKKEQEFTTNENN QTGVMIQVFEGERSMTCDNNLLGKFHLTGIPPAPRGVPQIKVTFDIDANGILTV SAADKSTGKTEHVTITNDKGRLSQQDIDRMVAEAEKFREDDEKKKRCVESKNEL ENYCYSMKNALEEEGVKSKLSSSELSEAQKLLQNTFSWIESNQLAEKEEFEAKLKE VQAVCTPLTAKLYQAGGGVPGGAAPGGFNAGGAAPSGPTVEEVD 22 MKLIACTLKNVETCVEVDPSDTVDALTNKIGSSLNNASASKMRLIHAGKILKME BMR1_03G04110 QKISDYSDIKDGDKIIVLFSKQSEASTIANPTPAPTSTPIADANTSPPKPIPTTDPNA full sequence LLMGEELEKAINGIVEMGFDVESVKAAMSAAFNNPNRAIELLTRHEVDVSDHD THQSVQTTGVLDELRQHPIVIFEQMRAIVRSNPQTLPQILSLIGQSDPSLLQAITE NQEEFIQLLSEPVLGTSGDFIDAQSITLTPEEMESINRLEGLGFSRPAAVEAFLAC DKNEEMAANYLLENIADYVSDNDN 23 MDGQTEQQLIEENIERFRILYQLHLDNNEPSPTAQFNYACALVCSNQRSHNDTA BMR1_02G02560 IYLLDELVRIRYESEECFYQLALAHMKRRSFVKSKEYLDRIIALEGSNQRVMALKSV full sequence WSLLAQDTFMGGLLGATAAFAIILFFTMKRNT 24 MNQNSLCCSKGFTMFAGGAFFLVSFKPETVITNPLLLRPLLNVSWGYIFGSHLW BMR1_04G05635 AAISTYNKKYWENRIIPDYANSPSREIMQSRLKINESRIYRIYLENLIQTNVLANGIL full sequence LVTTSALAPSNKFLRICSGAALLLSIGNWFALPEDEKDDDVGVVKTSSTSCFLSEV LSFCTFGAIVPYVFA

    [0111] In some embodiments, the compositions include one Bm antigen or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 1, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 2, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 3, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 4, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 5, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 6, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 7, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 8, or one or more antigenic fragments thereof. In some embodiments, the composition includes a Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 9, or one or more antigenic fragments thereof. In some embodiments, the composition includes one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3. In some embodiments, the composition includes one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof. In some embodiments, the composition includes one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0112] In some embodiments, the compositions include one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and also include one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    [0113] In some embodiments, the compositions include two Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof. An exemplary composition includes the following two Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1) and BMR1_04 G05532 (SEQ ID NO: 2). In some embodiments, the compositions include one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following two Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1) and BMR1_02 G01760 (SEQ ID NO: 4). In some embodiments, the compositions include two or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments of each of these two antigens. In some embodiments, the compositions include two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or two or more antigenic fragments thereof.

    [0114] In some embodiments, the compositions include three Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these three antigens. In some embodiments, the compositions include two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, and one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following three Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2) and BMR1_02 G01760 (SEQ ID NO: 4). In some embodiments, the compositions include one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following three Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_03 G00365 (SEQ ID NO: 5) and BMR1_03 G04695 (SEQ ID NO: 6). In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments of each of these three antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0115] In some embodiments, the compositions include four Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include four Bm antigens that each comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these four antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and at least one Bm antigen comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition include the following four Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_03 G00365 (SEQ ID NO: 5), and BMR1_03 G04695 (SEQ ID NO: 6). In some embodiments, the compositions include four Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments of each of these four antigens. In some embodiments, the compositions include four Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0116] In some embodiments, the compositions include five Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include five Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these five antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following five Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), and BMR1_03 G04695 (SEQ ID NO: 6). In some embodiments, the compositions include five Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include five Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0117] In some embodiments, the compositions include six Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include six Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these six antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following six Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), and BMR1_03 G03430 (SEQ ID NO: 7). In some embodiments, the compositions include six Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include six Bm antigens that each comprise an amino acid sequence having at 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0118] In some embodiments, the compositions include seven Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include seven Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these seven antigens. In some embodiments, the compositions include seven Bm antigens that each comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments of each of these seven antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and four Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and at least one Bm antigen that comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7) and BMR1_04 G06070 (SEQ ID NO: 8). In some embodiments, the compositions include seven Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0119] In other embodiments, the compositions include eight Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include eight Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments of each of these eight antigens. In some embodiments, the compositions include eight Bm antigens that each comprises an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments of each of these eight antigens. In some embodiments, the compositions include three Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and five Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. An exemplary composition includes the following eight Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_01 G00985 (SEQ ID NO: 3), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7), and BMR1_04 G06070 (SEQ ID NO: 8). In some embodiments, the compositions include two Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and six Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include eight Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0120] In other embodiments, the compositions include nine Bm antigens, one or more antigenic fragments thereof, or a mixture thereof. In some embodiments, the compositions include nine Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof. In some embodiments, the compositions include the following nine Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_01 G00985 (SEQ ID NO: 3), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7), BMR1_04 G06070 (SEQ ID NO: 8), BMR1_04 G09385 (SEQ ID NO: 9). In some embodiments, the compositions include nine Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0121] In some embodiments, the compositions include one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, or 24 or more) Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof. In some embodiments, the one or more Bm antigens each comprise an amino acid sequence having at least 90% sequence identity (e.g., at least 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof. In some embodiments, the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity (e.g., at least 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof. In some embodiments, the one or more Bm antigens each comprise an amino acid sequence having at least 99% sequence identity (e.g., at least 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof. In some embodiments, the one or more Bm antigens each comprise an amino acid sequence of any one of SEQ ID NOs: 1-24. In other embodiments, the compositions include twenty-four Bm antigens or twenty-four or more antigenic fragments thereof. In some embodiments, the compositions include twenty-four or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or twenty-four or more antigenic fragments thereof. In certain embodiments, the compositions include the following twenty-four Bm antigens: BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), BMR1_01 G00985 (SEQ ID NO: 3), BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7), BMR1_04 G06070 (SEQ ID NO: 8), BMR1_04 G09385 (SEQ ID NO: 9), BMR1_03 G04485 (SEQ ID NO: 10), BMR1_03 G01645 (SEQ ID NO: 11), BMR1_03 G01960 (SEQ ID NO: 12), BMR1_04 G05080 (SEQ ID NO: 13), BMR1_03 G00820 (SEQ ID NO: 14), BMR1_01 G02100 (SEQ ID NO: 15), BMR1_04 G07360 (SEQ ID NO: 16), BMR1_02 G02185 (SEQ ID NO: 17), BMR1_04 G08260 (SEQ ID NO: 18), BMR1_03 G02345 (SEQ ID NO: 19), BMR1_02 G02960 (SEQ ID NO: 20), BMR1_01 G02545 (SEQ ID NO: 21), BMR1_03 G04110 (SEQ ID NO: 22), BMR1_02 G02560 (SEQ ID NO: 23), and BMR1_04 G05635 (SEQ ID NO: 24).

    [0122] In some embodiments, the antigenic fragment may comprise a portion of the Bm antigen of at least 10 (e.g., at least 10, 25, 50, 75, 100, 150, 200, 500, 600, 700, 800, 900, 1000, or more) amino acid residues of a Bm antigen described herein. In other embodiments, the antigenic fragment may comprise a portion of the Bm antigen of at least 100 (e.g., at least 100, 150, 200, 500, 600, 700, 800, 900, 1000, or more) amino acid residues of a Bm antigen described herein. In further embodiments, the antigenic fragment may comprise a portion of the Bm antigen of at least 500 (e.g., at least 500, 600, 700, 800, 900, 1000, or more) amino acid residues of a Bm antigen described herein. Antigenic fragments may be of any length sufficient to induce an immune response when administered to a subject.

    [0123] Exemplary Bm antigens useful in conjunction with the compositions, methods, and kits described herein include those for which the amino acid sequence displays at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to one of the foregoing sequences, and in which the sequence changes are conservative amino acid substitutions. Further Bm antigens useful in conjunction with the compositions, methods, and kits described herein include those for which the amino acid sequence has one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-50, 2-40, 3-30, 4-25, 5-20, or 10-15) conservative amino acid substitutions with respect to one or more of the foregoing sequences.

    [0124] In some embodiments, the Bm antigens used in the compositions of the invention may be purified from the parasite or artificially manufactured molecules, e.g., recombinant proteins, synthetic peptides, proteins or peptides encoded by DNA plasmids or produced by recombinant viruses or bacteria. In some embodiments, the Bm antigens, antigenic fragments thereof, or nucleic acid molecules encoding them (e.g., RNA or DNA molecules) are in purified or isolated form. In some embodiments, the Bm antigens or antigenic fragments thereof, are at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% free of other molecules with which they may be naturally associated in nature. In some embodiments, the compositions of the invention are pharmaceutical compositions and do not exist in nature even if specified in general terms, e.g., a composition comprising the antigen or antigenic fragment and a pharmaceutically acceptable carrier or diluent.

    [0125] A composition may comprise one or more Bm antigens or antigenic fragments together with other components such as an excipient, preservative, diluent, or carrier. Exemplary excipients include sugars and sugar alcohols, such as lactose, sucrose, glucose, mannitol, and sorbitol, as well as gelatin, cellulose, cellulose derivatives, polyvinylpyrrolidine, starch, and polyethylene glycol. Exemplary preservatives include thimerosal, 2-phenoxyethanol, and formaldehyde. Exemplary diluents include water, saline (e.g., phosphate-buffered saline), and sucrose solutions. Exemplary carriers include human serum albumin.

    [0126] A composition may further comprise one or more adjuvants, as readily understood by those skilled in the art. Suitable adjuvants for compositions of the present invention include adjuvants that are capable of enhancing the immune response to the Bm antigens of the present invention (e.g., Bm antigens described herein). Adjuvants are well known in the art (see, e.g., Vaccine Design—The Subunit and Adjuvant Approach, 1995, Pharmaceutical Biotechnology, Volume 6, Eds. Powell and Newman, Plenum Press, New York and London, hereby incorporated by reference).

    [0127] Exemplary adjuvants for use in the compositions of the present invention include aluminum salts (e.g., aluminum oxyhydroxide and aluminum hydroxyphosphate) and calcium salts (e.g., calcium phosphate). A well-known example of aluminum oxyhydroxide is Alhydrogel™, whereas a well-known example of aluminum hydroxyphosphate is AdjuPhos™.

    [0128] Another adjuvant for use in the compositions of the present invention is an emulsion. An emulsion can be a water-in-oil-in-water emulsion or a water-in-oil emulsion. The oil phase may consist of squalene and squalane mixed in one of several commonly used ratios. A well-known example of such adjuvant oil is Montanide™ ISA-720 (Seppic, Castres, France), which contains both squalene and squalane, with squalene predominating. In addition to the water phase and the oil phase, emulsions contain a surfactant emulsifier. Examples of surfactant emulsifiers include mono- and di-fatty acid (C.sub.12-C.sub.24) esters of sorbitan or mannide, such as sorbitan monostearate, sorbitan monooleate, mannide monostearate, and mannide monooleate. An exemplary water-in-oil emulsion used in the composition of the invention may consist one or more Bm antigens or antigenic fragments thereof dissolved in the water phase and emulsified in Montanide™ ISA-720 using mannide monooleate (Arlacel™ A) as surfactant emulsifier. Oil-in-water emulsion adjuvants include those disclosed in WO 95/17210 and EP 0399842, incorporated herein by reference. An exemplary oil-in-water emulsion is MF59 which uses squalene as adjuvant oil (Chiron Corp; e.g., see, U.S. Pat. Nos. 5,709,879 and 6,086,901).

    [0129] Small molecules can be used as adjuvants. Such adjuvants include 7-substituted-8-sulfo-guanosine derivatives and 7-substituted-8-oxo-guanosine derivatives (e.g., 7-allyl-8-oxoguanosine (loxoribine)), as described in U.S. Pat. Nos. 4,539,205; 4,643,992; 5,011,828; and 5,093,318; which are each incorporated herein by reference.

    [0130] Lipid products of bacterial origin also can be used as adjuvants, including a monophosphoryl lipid A (MPL) (Corixa Corp.; see, U.S. Pat. No. 4,987,237), and a 3-O-deacylated derivative of MPL (3D-MPL). Cell wall proteoglycans of bacterial origin that can be used as adjuvants include muramyl dipeptide analogues (e.g., as described in U.S. Pat. No. 4,767,842), such as N-acetyl-muramyl-L-threonyl-D-isoglutamine (Thr-MDP; U.S. Pat. No. 4,606,918), N-acetyl-nor-muramyl-L-alanyl-D-isoglutamine (nor-MDP; CGP 11637), and N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanyl-2-(1′,2′-dipalmitoyl-sn-glycero-3′-hydroxyphosphoryloxy)-ethylamide (MTP-PE; CGP 1983A).

    [0131] Other exemplary adjuvants include the saponin fractions derived from the bark of the South American tree Quillaja saponaria Molina (e.g., Quil-ATM). Derivatives of Quil-A and methods for its production are described in U.S. Pat. No. 5,057,540. In addition to QS21 (also known as QA21), other fractions such as QS17 (also known as QA17) are described.

    [0132] Other exemplary adjuvants include RC-529, CpG oligodeoxynucleotides (Pfizer), SBAS2 (SmithKline Beecham), GM-CSF, Complete Freund's Adjuvant (CFA), and Incomplete Freund's Adjuvant (IFA). Yet another class of exemplary adjuvants consists of glycolipid analogues such as N-glycosylamides, N-glycosylureas, and N-glycosylcarbamates, each of which is substituted in the sugar residue by an amino acid.

    [0133] Adjuvant mixtures that can be used in the invention include, e.g., combinations of 3D-MPL and QS21 (see, e.g., EP0671948 B1), oil-in-water emulsions including 3D-MPL and QS21 (see, e.g., WO 95/17210 and PCT/EP98/05714), 3D-MPL formulated with other carriers (see, e.g., EP 0689454 B1), QS21 formulated in cholesterol-containing liposomes (see, e.g., WO 96/33739), and immunostimulatory oligonucleotides (see, e.g., WO 96/02555). Alternative adjuvants include those described in, e.g., WO 99/52549, and non-particulate suspensions of polyoxyethylene ether (see, e.g., UK Patent Application No. 9807805.8).

    [0134] Adjuvants are utilized in various amounts, which can vary depending upon the type of adjuvant, the components of the composition (e.g., compositions including one or more Bm antigens or antigenic fragments thereof), and the subject to which the composition is administered. Typical amounts can vary from about 1 μg to about 50 mg per dosage. Those skilled in the art can readily determine appropriate concentrations and amounts to use.

    [0135] Babesia microti Nucleic Acid-Based Compositions

    [0136] Compositions of the invention, which can be used, e.g., in the methods and kits described herein, can include one or more nucleic acid molecules (e.g., DNA or RNA (e.g., mRNA)) encoding one or more Bm antigens (or one or more antigenic fragments thereof) and optionally a pharmaceutically acceptable adjuvant, carrier or diluent.

    [0137] The nucleic acid molecules can encode any one or more of the Bm antigens or antigenic fragments described above in the subsection entitled “Babesia microti antigen-based compositions” or elsewhere herein. Accordingly, the nucleic acid molecules can encode any one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, or 24 or more) of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof (e.g., a fragment of at least 10, 25, 50, 75, 100, 150, 200, 500, 600, 700, 800, 900, 1000, or more amino acids). The nucleic acid molecules can further encode polypeptides having at least 80%, 85%, 90%, 95%, 97%, 99%, or greater sequence identity to any one of SEQ ID NOs: 1-24, or antigenic fragments thereof, consistent with the description provided above.

    [0138] In some embodiments, the nucleic acid molecules (e.g., RNA or DNA molecules) are in purified or isolated form. In some embodiments, nucleic acid molecules (e.g., RNA or DNA molecules) are at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% free of other molecules with which they may be naturally associated in nature. In some embodiments, the compositions of the invention are pharmaceutical compositions and do not exist in nature even if specified in general terms, e.g., a composition comprising the nucleic acid (e.g., RNA or DNA) and a pharmaceutically acceptable carrier or diluent.

    [0139] The nucleic acid molecules can comprise DNA, RNA (e.g., mRNA), or modified forms thereof. In some embodiments, the nucleic acid molecules comprise DNA that is designed to encode and express an RNA molecule that, in turn, encodes a Bm antigen or antigenic fragment thereof as described herein. Such DNA molecules can optionally be present in the context of a vector (e.g., a plasmid or an expression vector) that is administered to a subject.

    [0140] In some embodiments, the nucleic acid molecules comprise an RNA molecule that is administered to a subject. In some embodiments, the RNA comprises one or more modified nucleosides (e.g., pseudouridines and/or 2′-O-methylated nucleosides). In some embodiments, the RNA molecule is non-replicating RNA. In some embodiments, the RNA is self-replicating. In some embodiments, the RNA is present in a lipid nanoparticle. In some embodiments, the RNA is present in an RNA virus (e.g., a retrovirus, lentivirus, alphavirus, or rhabdovirus). In some embodiments, the RNA is introduced into dendritic cells ex vivo, and the dendritic cells are then administered to a subject (e.g., a subject from whom the dendritic cells were obtained).

    [0141] Nucleic acid molecule sequences encoding each of SEQ ID NOs: 1-24 are provided below in Table 2 as SEQ ID NOs: 25-48, respectively. DNA coding sequences are shown. As is well known in the art, in the case of RNA molecules, each “T” is replaced with a “U” or an equivalent nucleotide (e.g., pseudouridine). Also as known in the art, nucleic acid sequences can be codon optimized for use in the species in which expression is desired.

    TABLE-US-00002 TABLE 2 Nucleic acid sequences encoding exemplary full-length B. microti antigens or their ectodomain SEQ ID NO. Nucleic Acid Sequence Gene ID 25 GATGTATATGAGATATCTTCTGGTAATCCACCCGACATAGAGCCAACATCTA BMR1_01G03280 CTTCTCTAGAAACAAATGTAGTTACCAACTATATTCCAGAACCCAATGCGGAT ectodomain TCAGAATCTGTACATGTTGAAATCCAGGAACATGATAACATCAATCCACAAG ACGCTTGCGATAGTGAGCCGCTCGAACAAATGGATTCTGATACCAGGGTGT TGCCCGAAAGTTTGGATGAGGGGGTACCACACCAATTCTCTAGATTAGGGC ACCACTCAGACATGGCATCTGATATAAATGATGAAGAACCATCATTTAAAAT CGGCGAGAATGACATAATTCAACCACCCTGGGAAGATACAGCTCCATACCAT TCAATAGATGATGAAGAGCTTGACAACTTAATGAGACTAACGGCGCAAGAA ACAAGTGACGATCATGAAGAAGGGAATGGCAAACTCAATACGAATAAAAGT GAGAAGACTGAAAGAAAATCGCATGATACTCAGACACCGCAAGAAATATAT GAAGAGCTTGACAACTTACTGAGACTAACGGCACAAGAAATATATGAAGAG CGTAAAGAAGGGCATGGCAAACCCAATACGAATAAAAGTGAGAAGGCTGA AAGAAAATCGCATGATACTCAGACAACGCAAGAAATATGTGAAGAGTGTGA AGAAGGGCATGACAAAATCAATAAGAATAAAAGTGGAAATGCTGGAATAA AATCGTATGATACTCAGACACCGCAGGAAACAAGTGACGCTCATGAAGAAG GGCATGACGAAATCAATACGAATAAAAGTGAGAAGGCTGAAAGAAAATCG CATGATACTCAGACAACGCAAGAAATATGTGAAGAGTGTGAAGAAGGGCAT GACAAAATCAATAAGAATAAAAGTGGAAATGCTGGAATAAAATCGTATGAT ACTCAGACACCGCAGGAAACAAGTGACGCTCATGAAGAAGAGCATGGCAAT CTCAATAAGAATAAAAGTGGGAAGGCTGGAATAAAATCGCATAATACTCAG ACACCGCTGAAAAAAAAAGACTTTTGTAAAGAAGGGTGTCATGGTTGCAAT AATAAGCCCGAGGATAATGAAAGAGACCCGTCGTCGCCTGATGATGATGGT GGCTGCGAATGCGGCATGACGAATCACTTTGTCTTTGACTACAAGACAACAC TCTTGTTAAAGAGCCTCAAGACTGAAACA 26 ATAATTACGGATGGATTGTCCGCTATTGGCTCTGTGGCGAGTGAAGTGGGA BMR1_04G05532 AATACAGTGAAGGATGTTTCCAGTGAAGCGCTCTTAGGAGAATTACAACAA ectodomain ATTGCAGACGGTGGCAAAATTATAAGAGACGGCACTGAAAGCAATTTTGTG AATTCAATAGCTAATACGGTTATGAAAAATGTAGTTGGCACTGCAGTTCTCA AAGCTTCATCTGGTATTACACATAATGGTGATTTCGCCTTTTACAACTTTATG TACCCGGAAAATGATGATTATCCATGGGCATGTATCTGTGATGAATCTGATT ATGAAGAGTATATTAAGGGCAAGAAGGACAAGGTTAGGTGCAGGAACTAT ATTGATTCATCCTTACAAAATGCAGTTTTATATTGTAACCCAGCAAATCATAA CTCTTCCATAAATGACAATGCCAATAACAATCCCCCAAAGCAGATTGATAAC CATGTGTCCATACCACAAACAGCGCCAGCAAATCATACCACAGTTTTATCTAC GGAAGTCGACACTAATCACAATGAACAAAAACAACCAAATTCGCCTTCAGTT CCAAGTGAATCTCAAAATTCAGTGTCCGCTCCGAAAGATGAATCTGTTAGTA GCACCGTAGAAGGAGCCAAATCAAGTTCA 27 ACACTTGTATCTAAAATTACAACTCCAAACAATTTACCCGGAATAGATACTTG BMR1_01G00985 CTCGAACTGGGAAGATGTATCAGTGTGTACTACTAAAAACACCAGAAATTGC ectodomain ATTTCAGGTGAAGGCAAACCAAAGGATTGTTTTGCGATTGGGAAATCCTTAT TTAAAACATTTCCAAATTGTTATGAAGGTGTTATAATTGATCAAGTCACGTTT TCTGGATTTGAGACACTAGAAATCCATTATTGCGATATAGATAAGATCTTAC ACAAAGCAAATGAAGTTGTTAAATCAATACACGAATTGAAAGAGAAAACAA ATCAATTAACCGAGAAAATTAAGGACATACCTGATTTGATTGATAAAGTTAT TAAGGTAAATTCGGAAATTTCAAAAATATTTCATCAAGATAAAATTAAACAT ATGGAAAAGGAGGCCAACGATTTCAAAAACGCCTTGAAAACCACTAGAAAC TACATAATCAGTTATGATTCCTTAGATAAAACCAAGCAAAGCAATCTACTAAC TTCACTTGGAAAATTAATGAATAAGATTAAAACAAAACTATCAGAAATGGAT AAGACATTACATTCTACACTTGACACAAACAATACCATTATAGATCTTGTCAA TAACAATAGTTCACACGTTAAACATCCTAATGATTTTAACAAAACAATGGAA CTCTACAATGAAACTATTACTAAAGCTGATGCGATTAAAAAAAATATTGAAA AACTTAAAGAACATAGGAAAATATCTACTCACAAAACAATATTTTCAAACAA TATAGATAAGTTGATTGACAATTTGACAGATTATTTTGAAAATATAAATCGCT CCATTGATGCTGTTAGAGACAAACTTTCCAAATACCAACTTGAAACGGGCAA GATGGTTTTATTGTTTAAGAACGTGAATGAAATTCAAAAGCATATTAAAAAT ACAGATATGCATATAAGAACGTGCAATTATGATTTCTCAGATATCGAACAAA AATACTCACTCATTACTGCAAAAATTACTGTCGAAGATGGACAATCAATCAC AACTTCAAATAAATCAACTGTGGATATACCAGAAGAGAAGGTAGATAGGGT GAATGTCAATGTTGACAAAGCAGAGAACTCTGATAATGAAACTTCTCAGGA AAATACATCTGTTAAACCGACTGATCACAAAGAAATTGAGGATTCAGCTTCC GAAGAAAATGCCATTGGAGAAAATGGTGATTATGATTCAGATGAAGATATT GATACAAATGACGTTAAAGAAGATCACGAAAATGCAATTGATTCCGAATAC ACCGTTTCATCGACTGGGGATGTATTAGAAGATGAGGTAGTTGAGGAAAAT GCCATTGGAGAAAATGGTGATTATGATTCAGATGAAGATATTGATACAAAT GACGTTAAAGAAGATCACGAAAATGCAATTGATTCCGAATACACCGTTTCAT CGACTGGGGATGTATTAGAAGATGAGGTAGTTGAGGAAAATGCCATTGGA GAAAATGGTGATTATGATTCAGATGAAGATATTGATACAAATGACGTTAAA GAAGATCACGAAAATGCAATTGATTCCGAATACACCGTTTCATCGACTGGGG ATGTATTAGAAGATGAGGTAGTTGAGGAAAATGCCATTGGAGAAAATGGTG ATTATGATTCAGATGAAGATATTGATACAAATGACGTTAAAGAAGATCACGA AAATGCAATTGATTCCGAATACACCGTTTCATCGACTGGGGATGTATTAGAA GATGAGGTAGTTGAGGAAAATGCCATTGGAGAAAATGGTGATTATGATTCA GATGAAGATATTGATACAAATGACGTTAAAGAAGATCACGAAAATGCAATT GATTCCGAATACCTCGTGTCATCGACTGGGGATGTATTAGAAGATGAGGTA GTTGAGGAAAATGCCATTGGAGAAAATGGTGATTATGATTCAGATGAAGAT ATTGATACAAATGACGTTAAAGAAGATCACGAGGATGCAATTGATTCCGAAT ACCTCGTGTCATTGACTGGGGATGTATTAGAAGATGAGGTAGTTGAGGAAA ATGCCATTGGAGAAAATGGTGATCATGATTCAGATGAAGATATTGATATAG AAGAAGTTAACGAAGAGGATCATGAAGATGCGATTGATTCTGATAACTCCA TATCAAATAGCGAAAATGTAGATACTACACCAACTGAGAATGTAGATACTAT ACCAACTAAGAATGCAAATACTACACCAACTAAGAATGCAAATGCTACACCG ACTAAGAATGTAGATACTATACCAACTAAGAATGTAGATACTATACCAACTA AGAATGCAAATACTACACCAACTAAGAATGTAGTTACGACACCAACTAAGAA TGCAAATACTACACCAACTAAGAATGTAGTTACGACACCAACTAAGAATGCA AATACTACACCAACTAAGAATGTAGTTACGACACCAACTAAGAATGCAAATA CTACACCAACTAAGAATGTAGTTACGACATCAACTGGAAATGTAAGAACCAA ACACACATCGACAAATTCTCATGTTTTAGCTCCGGATACAGACGAATACCCA GCACAAATTTCACAACATAAGACGATTGATAAATATTATCAATCGTTGGAAT TGGAGGATGAAGAAAATGCAGATATATCAAGCTCAGATAAACCAGTTTCTCC AATAAATTTGGAAGATAAAAATTCGACATATGATCATATGCATAAAACCGAT AATGTTAAAAGCGCCGGTATTGCTTCATATGAT 28 AACCGCTCTTGTCCAGGAAACAATGGCGTTGGCGGTGGATCTGGTGATAAT BMR1_02G01760 AACAGTGGCATAATTCCAAATGATCCACACCCCTGTTGTAACAATCTTAGAC ectodomain AAAAACCCCAATACCAAACCAAGCCAGAAAATGAACTAGTCAATGATGATA GAGATTTGAACTTTAATAAGATCAGAGGTGGCAAGCAAATCATCACCTTTAC TGTCCCTTCCATAGATGATCTCAAGAATAAAAGATTATCCGACTCTGAATTCA TTTTATCAGAAAAGGCAAATCCATTGATTTCCTCCGGCGACAGTAAAAACGT TATTGTATTCGAAGTAAAAAACGATAATGAGAAGTTAATGGGTAGTGTTGA AGTTGGTCAATGGGAAGTTACTATCACCACATCATGCATCAGACGTATCGTT ATTTTCGATTCCAATGAAGTTTCAGATAACATTCCCATGTATATATATATCGT GGACTACTTTGAAGGAGGTAATAGCACTGTTTCAAAATTCTTTTTCGCGAAT AATAGATGGAACGCTGATTTCACCAATCACACACCTAATGCTGCT 29 TTCCTACTGAACCGCAGCGAATTTAAGTGGTTTAAAGTGGGTCTAATAATTA BMR1_03G00365 CCACGATATTCCCTTTTAAGCATTCATTTGACTATAATTTGACACACATATTTC ectodomain TATTTTCAATATGCACGTTAATATTTTGTGTAAAACCTGTGGATGAGGAGAG CGGTGCCAAAAAGGAGGGATTTGACTTCAAGAAGATGGTGCCAGATAAGTT CAAAAAGTATACC 30 GATAGTGGTAATTCTTCCCCCCAAACGCCGCCAGAAACTAGCAGTCCTATAA BMR1_03G04695 ATGGTGTAATCGGCGATGAAAATAATGGATTGGAACATTTGAGTAGTTCTG ectodomain GTCTGGAAGTTGATGATAACCTGCCCGAATTACTCAAAACTTCTCCATTTTCA GGCCAAAATTCAGATGTTCAATCTGCTTCAACACCAGTTGAACCCACTACTCC TGTTCATTCCAATGACCAATCTAATCCTATCACCAATAAAGTTGATACCAATT CTAATGACCATACTGATATTAAGAATGAAGGTTCATCCCATCGTACTTCATCC AACAATTCTTCTGTCACAACCAATACTAATAATGAGATTAGAAATGGCGGAG GACCATTAGATCAGAATGAAGATAAGGCTGAAGATGAAGGTGAAACTGATG CCGAAGGGAGAGGATGGAATGAGAGAACGAAAAATAAACCTACATTTAAT GCCACAAATCGCGATTTTGTTGATGATAACCTGCCCGAATTACTCAAAACTTC TCCATTTTCAGGCCAAAATTCAGATGTTCAATCTGCTTCAACACCAGTTGAAC CCACTACTCCTGTTCATTCCAATGACCAATCTAATCCTATCACCAATAAAGTT GATACCAATTCTAATGACCATACTGATATTAAGAATGAAGGTTCATCCCATC GTACTTCATCCAACAATTCTTCTGTCACAACCAATACTAATAATGAGATTAGA AATGGCGGAGGACCATTAGATCAGAATGAAGATAAGGCTGAAGATGAAGG TGAAACTGATGCCGAAGGGAGAGGATGGAATGAGAGAACGAAAAATAAAC CTACATTTAATGCCACAAATCGCGCTTCTCCCGATGGAATTGGCAAAATGAA CATGGAAGAAAAACAGTTGGAGAATTTTATAAATGTATCATCCAACGCTCTT GAATTAGATATCTCAATTGGCCGGGATAATTTTGCTACTAAATTCCTAGCACA ACACGTTAATATTTTTGGAGACAGGATAAGCGGCTTGAGTGCGGCGTATGT AGAGGGCTATAATAACTTGGCTAAAATTATGTACAATAGTCACTCTGTTTTAT TTGATAGAAAATTCAATGGAGCAGTTATTTCCGACAATTTGATTGGAAATAT TGCAGATTTTGGTTCATACTTCTTAGAGATATCTCCAAATACAACTAGAACCA ATAGAAGTGATTATCTCAAATCAGTAGTACTTTCCAAAGTTCAGTATCTGTTG TCTGCTGATTTTTCCACCACTGACAACATCCAAAGGCTCACAAATTTGGCACT TGCCCTAGGCTATAACAATGTCAAAGAAAATAATCCTGGCAATTCCCAACAT TCAATAACAACCTCCCTCTCAACTGAGCTATTTTGGTCCTTTGGCAACAATAT ATTCTTATTTGGACACTTGGCAACTCTAATGCTTGCTTATCTAGAATCTAATG CATATTTCACATCTGGTGCGACCAGGCCTTTCTTTTCATGGCAAACTCTGGTT TCCACTGGTGGTAATGAGAAATTTGATAAACTGGACTCTATGTGTGGAGTGA TTCGTGGATCAAAGTATTCGCGAAAAAACAATGGATTTATCAAGCCACACTA CAAAAGATTACGAAGGAAGACTTTGCTCGAAGGTGAACCGCGTTTGTTGTG CAGCATGCTGGAAGAGGCTTTGGATACTGTGGATAAAGCTATTAAATTTAA GGGCGAAGAGCTAAATTCACAAGGCGCAAATATTGAAAATTCGGTCTCCAA TGACATCAACAGTAAACGTTTACAGGCGAAACTTTGCTCAAATTTGAATGAT TCACTTATAAACGTGAGCTGTGATTTTAGATCATCAAAATTGGACAAACACA ATAAGAAATTAAGAGAGGCATTTGACTTATTATTAGCTTGTGGCAATTTGAA CACTGGTAAAAAGGAGGCATTTCCGGAATACTTGAGATTGATCTCTAACCCC TTCGAATACGGCGATATTTTTTCCATGACTATGTGGTGGGATCCTAGGGAGT TTGATGGCAAACAGGGCTGGGTCGAAATATACAAAAAGTTGAGGAAGAATA TTATGAAACCTGAACTGAAAAATGTGGACATGCAACTCAAATACGACTCGGC AATTTCTTATTACAAACAACTGAAGGAATCGGAAACCTACCCCAAGAAAAAC ATTCCATGGGCTAGATTATACCTTTACATGTCAGTAATCATGTCAAGATCGAA CGCTATGAGTTGGGCTGAGGACGCCCTCCGCTCATTTAGCAATCTATATCGC ATGAAACCTTCATTAGTGATGAGAGGGGAAGGGTTAGAGACTTTGCTCAAT TACTGCGCTCCTGATCCAGTGGCTTTATCACACATATTCCTCTACCACTTTTTG ACAAAGAAAGATGCCGGTAAAGATTTGGAAAAAGACTTGAGAAGACTTGA GAAAGGGACACTCCTCTCGCGCATTGTGAATTCATCGTCAATATTCATACCTA ATAAACTAAAGAAATTCCTTAAAATGGGCGCTCGCGGATTTTTCAATAAAAA ACTTAACACTTTGAGAGCAAAGAGCACCCTTCTACGATTATTCCCCAAGAATT TACTACATTCTGCATTGGGCGCAATTGAATTCACCACACATTCACTAGCCACG TTACAAATATCTAAGAATATGGATATGTGGGAGAGTTTGGCCCAGACAAAG AATTTAGATGCAGGCGGTTTCCCTGGCGAAATCGATTCACTGTTTAATCATT GGTCGGAAAGTGGTGGATACAGTGGGTATATTACCGGCAAATTAGAGAATG GAGATGATCTAACGGGCGATGATATCAAAAAAATGAACATAAAGGCCCCTA TTAACAATGATTCACTCAATTGGCAAAAGTATATCAATAAAAAAATATCCGA GCATTTTGGCAAATTTCTAAACCTTCCATTTATCCAAGCCTCCGGTTCACAAA AGAACTACATATATCAGCTTGTACGGGATAGTAAGGCAAACCTTGACGATAA TTTGGAACAAACTGTGTTCTTTGGCAAAGTGCTTCCCCCAGGAAAAACTAAT AATGTTATTAAGAAATTGAAAAGAATTGCAGATTCATTTACCAGCATGCTGT TACGTTCTTCTGCTCGTCCTGTTGACCATGCTGTGTGGGTTGGAGTGAAAAT CAACGTACCTATTGTAATTCATATTACTAAAAAGTTGTACATGATACAGCGTG ATATGCCTAGGAAGGAAGCGTGGAATTTGGAAAGTGCTTTCTTGGACCTGTT GCAGGATCTGGTGATAATGGTTACAAACCCGGGGAAGAGATCTCCGATAGG GTTTGAAACGATTGGGGGCAATCCTGGATTACCAGAAATTAGCATTAGATAC CCTCATATGTCGATTGAGGAAAGGAAGATTGAATTTCAACATTCACAATGTG CAGATCATTGTATATCAATTTGGAGATCCCTAATCGCATTTACACTTAACACA CTAAACAATCCTGCGGCAATAAAGCAATTTGAGAAATCCTTATCCAGTAATA GTTCTCTAAACGACATGTCGAAGCCCGAATATATCAACAGCTTCAAGTATAT ACTAAAAGGCGACTCGGTGTTGCATATGTATGATAACATGTTGCCTAGGAA GGTGAAAAGGGAAATAAAGGCACTAAAGTATGGTAAG 31 CTGCCTGACATTCTATCGCAGAATAACACGTTTAAATCTTTTCTCGAAGTAAA BMR1_03G03430 TAACGTGGATCAGGAAGATTTGATTTGTAATAAGGCACTGTGTAAATCCACT ectodomain GACTCTATCAACAGAAATACAAGTTCTTATTGTTATAAATACAAACTGTGTAG TAAGTGTAGCGTATCCAACGTCCCAGATCACCCTGTTTGCTATCTTCTGGATA ATGACCACAATTACATCCACTTAATGGAGGGGCATTTAGGCTCTCAGCCCAT AGGATCAGCGAATAGTCACGATAATTCTTCTCATGACGAACATTCATCGCAT TCGAATAATGGAGATATGATGGATGAGCATGAAGAGGAAAATTTTCTGCAG GAATACGAATCAAAATCAATGAAATTCATACCTACTAGCAACATGAGTGACT TTGATCATGCTAGGCGATCTTGTGCCGTGGATTCCAAAGGCAATGTAATGAT AAGTGTTAGATTGATCATCCAATGGTATATGTCAAAGGATAAATCTAATAAT CAGCAGCACCATGGCAATGATGATGATTCTCAAAATTATGACGCAAATTATT TACAACTCACCCCTATGTATTCTGACGACTCTGTTAATTCTTCTATGTTAGAG ATGGACCACGACGATAGTGAATCATCAAATTCGCATAAATCTAGGATGGCA AATATGGCCAAAAACTTCCAAGTTTTGAAAAACATCCATAAAAGTGCAGTTA AGCGTTATAAATCTCCAAAGGCCAAGATATATCTCATATTTTCTAATCCAAAG ATCAACAGCTGTAGACACCCGGTAATATACAACGGTAAAATCTCCCCCTCAA GCATGTTTGTAGCGAAACTTGAGTCAACAATATCACAAATTGATTTGACACA AGATCTGATCAAATCGTCAATAGAAACTATTGTCTCTTGTGAAGCTTGTGAT AAGTTAAAATACAACAGTTGTATACAGGTTACCTGTGCCAAAAATACACCAG GCGCAGCGTCTTTAGCTATGGGTAGTGCTGTATACGTTCCAATGACGAATAC TACGATTGGAGTAAATGCCCACAACCCTAATGCGGTAGTAGCTGCCGGTATC CCTATGGGTAAAATACCTGTAATCCCCCATCCAGCAGCTATAAGTGGTGGAA ATGTTGGTCATTTGAATAATGGTTTGCACAAGGCGGTTAATAATGCTGTTAT GATGCCAAATGGAACATCATTGCCAGTGCAAAGCGGAGTTGTTATAAAATC ATTATACAATTGTCTCGCTTTTTTACTGACAATTTTGTATCTCAATTTC 32 AACCCTATATTTGCATCTGCAACCTCAGCTCCAAGTAGGAATAGAGCTCAAG BMR1_04G06070 ACATGACCAAAGCGGAATGTATAGAACTATTAAAAGGCATAGTGAAATCTC ectodomain AAGAGGAAACCAAAATTGTTATGAAGAAGTTGACCTCAGACCTGATAAACA ATCCACTGCGCCTAGAACAGGTATATACCAAGGCCAGTCAAATGCAGCCAG AGGATCCTATGGAGCAATGGGGAGTCACTGTAATTGATCTCGACCATTTGAT TGAAAAGTATCAACATGATCCTATTGTGAAGGATTACATTTTGAAGATCATG AATTCGCCCGGCATAAACGATACAACTCTGAGTGACGATGTACGCAATATTA CTATTAGTCAAATATTAGCTATCCATGAATATATGCTCAGCGAATTGGAGTCT GTAGTCCGCGAATTCAAATCGCTGCAGAACAGACAAACTATGGAGATTAAA ACGTTAACTATTGCTGCGCAAGCAATCGTAGCTGCCAAAGTAGAGGAAAAA TTCAATTTAACATCAGATCAGGTTGAATCGGCTGTGATAATAAACCATGCCG AATTAACTGCTAGCCATGCATTCACCAGATTGACTATGCAAATGCAGACGGA GATGAGTGAGTTGATTGGGTGCCAATTTCCCGGGTGG 33 AGTACGAATGGTAAGGGGGGTGTAGACTCTGTGTCGAAGAAATCTTTCGTC BMR1_04G09385 ATTGAACTTGAAGACTCCACTTTTGAAAGAAAGACCCAAGCTTCTTCTGGTG ectodomain GCACTTCTGGTGTATGGTTTGTCAAGTTTTATGCACCATGGTGTGGACATTGT AGATCAATGGAGAATGATTGGAATGAGTTGGCCAATATTTTGGGCAAACAA ATTAATGTAGCAAAAATTGATGCTACAAAACACAGTGTTACTGCGAAAAGAT TTGGAATTACAAGTTTCCCAACACTTTTGTTGCTGAAAGATGGAAATTTCTAT CAATATGAAAATAATAATAGAACTGCTGATGCTCTTAAACAATTCGCTTTGCA TGGCTATAAACAGGTTAAATCAAAGCCTGTTCCTAAAGAATGGAGTTATTTT GTTCGGTTCAAATTCTTTATCAAATCTGGATTTTATGAAGTTAAGCGCATATA CCAGTTAGCATATCCTGGGTTCATAACA 34 ATGGCGAAATTACATGAGAGTACATCAAGTGCAAGTGCTAGCTTCGACCCA BMR1_03G04485 GAGAAGTCGGATTATGACGATACTTATGTTTTAACAGAGACAACGCCAACTT full sequence ACATAAGGCATGGATTTGTTCGAAAGGTCTTTGCTATTTTGTTTGCTCAATTA CTAGTGACCCTTGGATTTTCATTGATCTGTTATTTLTACCGAGAATCTGTTCAT TCGTTCATATCCAAAAATATCTGGATTTTTCCAACCTTAGCTATATTATCATTT ATAACATCCCTTATACTCATATTTTCACCTTCACTCTCTAGGAGATATCCGTTG AATTACGCTATTCTCGTTATAGAAACATTGTATTTCTCATTCATTGTCGGATTG TCATGCGCGTTTACTAAAAGTCCGACTGCCATAGTATTATCCGTCTCTATTAC TCTTGGGATCATTCTTCTTGTTGTTCTGTTCACACTCCAGACCAAAATTGACTT CACTAGATATATTATCTATTTTATTTTATTTAGTTTTGTAACGTTGGTTTTTGG CTTTATTGGTATCTTTGTCCCATTTGACACCCCTCTCAGGATGTTTTATTACGG TTTAGGCGTACTTGGTTATTCACTTTGGATGGTACTTGATCTTCAACTGATTA TCGGTGGCAAGACTTACGAATGGACGGTTGATGATTACGTCCCTGCATCACT CTCGCTTTATACAGATGTTATTGGAATTTTCTTAAACGTTCATGGCATGTTTTC TGATAGA 35 ATGACTATAACCTTGAATATAAAGGTTAATTCTGAGACAAATTTTACGGTAG BMR1_03G01645 AGGCTGAACCTTCCTTTACTGTAAAGGAATTAAAGATATTATGTGAATCACA full sequence ATCGAATATTGAGGCGCAAAATCAACGCTTAATTTGTAAGGGCAAGCTTTTG AAGGATACAGACATTCTTTCCGATGTAGGGGCAGTTGATGGTGCCACGGTTT ACCTGGTTCGTAGCCAAGTCAACAAAACGCAATCAGCTGCTCCTAAGCAAAA TACTGTCCCTCAACCCACATTACAGACTACAAATCAACCTGCTGGTCAAACAC AAACGTCTGGACTTGGATTTCAACAACAAGGGTTTCAGCAAGGATTTTCTGG ATCTCAGCAACCAGGATTTCAAGCTAACCCCTTCCAAAGCATGCTTGCTGGT GGATTTCCCAATTTAGATCCCACTCAAATGATGGAGATTTTGAATAGTCCAAT GGCACAGGAAGCTATGCAGAGATTGAGCCAGAATCCAGAAGTTTTGAGGAA TATTTTGCAGAACTCATCTTTGATGACTCCCATGCTTGAACAAAATCCTATGC TATCAGAAATGCTATCAAACCCGGAATTGATGAGAAGTATGTTGAGACCCG AGGTTTTACAAGCCGGATTGCAAATGCACCAAGCCATGCAGCAGCAGCAAC AACAACAACCTGGGACTCAAACCAATCCAATAGGTTCACAAAATCCAGACTT CAGCAACATGATGAGGCAAATGATGAATGTATTTCAGCAAAATCCATCTGTT GCACAGCCCACAGCACCACAGGTATATACCGACCCGAGGCCTCCGGCTGAA AGATTTGCAACTCAGCTACAGGCATTGGCTGAGATGGGATTCATTGACACTG AAAAAAATATCACTGCACTTATTGCGACCAATGGCGATCTCAACGCTACAGT GACCAGGCTGCTTGAATCCAATTTT 36 ATGGAAGAGGCTGAACGCAAATTCAAGGAATTTAATTGGGCCGATAGTCAG BMR1_03G01960 GGATGGCGAATTTACTGGGACAACTTGTATCCAACTCCCCCCTTATCCAAAG full sequence TAGATAAGTTCAAACGTTCCTGGTTCAAGAGAAACGTAGATTCTAATATATC TTCAACACCACTGTCTGAGGTTGGCAAACAAACTCAACAAACTACTCCAAAT CAATATCGTTCGCAGGGCAACTTTGCTATTTTAACTCTCGAGGCCGGTGTTA GATTGTTGTACCTGCTTATTACAGCACCTTTATTTGTTCTACCACTAATAGGA GTCAGGCTCAGTAGATATATATACTACCACTACTACATAGACATAGCGCTGT TATTGATCTTTCTATTGTCTGGCATCGTAAGGGAAAGAGGTCTACCGAAAAT GGATTCCGTGTGGCTGGCATCTGCTTTCTATTCAGACATGACACAATATATA ATGTATACATGCATCCTGATGATGTCCGCACCAAGGCCTGTTTATCTGATTTT ACCCTTTCTAACTTGTCTAATCGGGCTAAATTCTCTGGCCGAAACAAATTTAT CCAAACTGCCCAAATGGCTTGAAAACATAGTGGGTGAAATATTTAAGTATAC TAAGGATAATATATATTGGTTAATGCAAACTCGTGGCGATGTTGAGTGTTAT TTGTTGTTTTATATTGTATTCGGGTTCCTAACGAAATCAAGCGCAGTTATCAC TTTAATGGCCTACATAAACTTTATGAAGTTAAGGATTGGCATTGGTGATCCCT TTATAATGTCTGCCTTCTCTAAATTGCATGGCTGCATAGTGAAGTTGTTATCG TATGATATGTTTGGACATATACCTTTGAGGATGTACTTGAAGATATCAGAGC TCTTAACTAAATATATGTCGGGGCCAAGGCCGCAATAT 37 ATGGAGGACCAAAATACTACAAATGAAAGCATTTCAAATTTGCACATGGAA BMR1_04G05080 AACTACTTCCCCAAAGATATTTTTTCCAACCTAGATAATCAAAAAAACTTAAA full sequence AACATATCATTCAAAGACGTTTGAGAAATATTTCGAGTCTGCTGCCAAAGAT AATGTTGAAAGGTGTCGTACATCTGCTGTCAAAGAATGGCTGAATAGAAATC TTCCTAGTGAATATAATGAAAGGCACAAGCCCACACTTAAACTAAACCTGAG CCCCAACCACTTAAACTCAACTTACAACAAACAAATTTCCGATTACACAAGGA ATGCCTATAATCGCATAGAACAGTTGAAGAAGCTTCAAGAAGCTTACTCTCT GTTAAGACAGAAACTACAAGAGAAACGCAGGGCCTCTTGTCATAAGGAGGT CTCTGAGGCAGAGGCTTCAGTTCTTAATACTCAAAGATTGATAATTAGCTTA AAGAGAGAAGACAAAGATAAAATCGCCACAGAGTTACTTATACAAAATGCA GAAGAGTTAGGTATACCATCCAAAGATTTGGAAACTCTAAACGGCTACACAT TTAATGAGGCTCTCAAAACTATAATCGATATAATTTCTAACATGATAAACAAC AAATTTATGACAAATTTGAAAGATAGGTGTGAATCAGCTAGGCGTAAGGCA GGATCTGAATATCGCGATGAAATGGAAAGGATAAAAGTCGATTTGGACATG TACAAAACAAAGTCTGAAAAACTCGAATCCACAATATCTACATTGAATTCTTA TGCAGATTCTAGCAAACAAAATGCTGAAAAGGTCATGGAATTGCAGCATTC GCTAGAAGATTCCAATAGACAAATTAATCAGCTAAAAGATTCACTGTACAAT ATGGCCAAAGTTAATGAGGAGTTGGAAAAAAACGAAGTTAAGCTAGAGGA AAGTTTGGTAGAGTTGGAGAGGTATAAGTTGGAATCTCAAAAGCTTGAGTC AGTCATTAAAGATTTAGAGCTTTTGAACCAACAAAAACATGATAATGAAGAA CTCATTAAAATGCTGCATGATGAGCTAGATCAGTGTAAGTCTAAGTCACTGC AATTGAATAAAAAGATTGAAGATCTTGAAAATTCAAACAAGGAAAACCTAAT ACCCTTGAACAATGAATTGGCCCAGGCTTATCAGAAACTATCCGAATTGGAG CATTCATATGAACAGATTGAACACCAGAAGAATGAAGCAGATAAAAAACTT GAATCTTCAATTGATGAGATTGAGAAACAGAAACAACACTCCGAAGAATTG GAGCAATCCATCACTAAATTGAAACAAACAATTGAAGAAATGGAAAAGGAA ACCACGGACCAAGTAAATGATCTGCAAACGTGCCTAATAGATGCGAATTGTA AAATCGAATCGCTAGAGGGAACGATTTCTCAGTTGAAACAGCTAAACTTAGA GTTGGAAGGGGGAGAACATAGGATACAGGAATTACAATCCAAGCTTACCGA ATCAAATTCCACAATTGAAAAACTTGAAAAGACGATCACAGAATTGGAAAAT GTGTCTAGTAAATATATCGACTACGATGAGAAGATGGATAATTTGCAAAAGC AGCTGAAAGAATACACTGAAAAGATCACAGTGCTGGAAAACAACGCCTCTG AATTCAACTATAAAGACCAAGCTGAAACGTTGAAGACGGAACTGGATAAGT CGCAACTAAAGATAATGGAACTGGAGACTATGATTAAGGAGAACAGCGAAA AAACGGAAAGAGTGCCATCTCCCAGAAAGGACAACGAGGAGGTTGATAGA TTAACTTCGGAAATATTACAGTTGAAGAATCAGGTGGACATTCTGCAAAATG AAAAGACTGACCTTGAACAAAAATTATCACAACAATCTCCCAGAAAGGACAA CGAGGAGGTTGATAGATTAACTTCGGAAATATTACAGTTGAAGAATCAGGT GGACATTCTGCAAAATGAAAAGACTGACCTTGAACAAAAATTATCACAACAA TCTCCCAGAAAGGACAACGAGGAAGTTGATAGATTAACTTCGGAAATATTAC AGTTGAAGAATCAGGTGGACATTCTGCAAAATGAAAAGACTGACCTTGAAC AAAAATTATCACAACAATCTCCCAGAAAGGACAACGAGGAAGTTGATAGAT TAACTTCGGAAATATTACAGTTGAAGAATCAGGTGGACATTCTGCAAAATGA AAAGACTGACCTTGAACAAAAATTATCACAACAATCTCCCAGAAAGGACAAC GAGGAGGTTGATAGATTAACTTCGGAAATATTACAGTTGAAGAATCAGGTG GACATTCTGCAAAATGAAAAGACTGACCTTGAACAAAAATTATCACAAATTA GTGCGGTTATTGAAGAAAATAGACAATACAAGGAAAAAATCGAATTACTCG AGAGAAAATTGAATGAAATAAATAAAGAGAAACCAAAAGACTCATTTACAA ACGTAGAAGTTATCAATGCAGCTTTTGAAGATGTAAGATCCCTCCCAATCAA TTCCTCCAATGCCTCTGATTGGACCGCAATGCCCAGTGAGCTGGAGCTGGAA GAGCATAAATTGTACAAAAAAAAATCTAGTAGAAAGGGGAAAAAGAAATCA TCAAGGGAGAAGGAGACTAGCAGAAGTGACATATCATCCAGGTCAACTAGC AGGCACTCAAAGAAGGATAAGCCTACAATCGTTGACAACAACTCTACTGATG TAGAAGCATCAGATTCAAATGAACAGATGATTAGTGATCCAGTTGAATCGAT TACAGACCAGTTGAACTTGGTTAAACAATCAACAATACAACTAACTGAGCTG GGGTACGACAGCATATCCAACGTTGGGTCGTACTTGTCTGACTACATATTTG GCGGTAAT 38 ATGGATGATCTACCCGGTTCATTACATCAATCAACACCAAATGAGAAGCCAA BMR1_03G00820 TGGCTCCACCCACAGCACCAAAATCTGCTCCACACGTTCCTATTTCTGTCGAA full sequence TCAAAGGAGTTATCAAAAGAAATTCCAAAAGAATCTCCTATGACTAAGGAG GACAAGAAAGACGTGGCCAAGAGTAAAGTTTCTGCTGCTGTAACTGAGAAA AAAGTTGTTAAGGAACCAGTGGTACCTAAAATAACAATTGAGATGAAAAAA TTTTCTATGTCCAGAGAATCCACCCAATACTCAATCTTCATTATATTTAACCTT ATCTTTGCACTATTATACATTTTCAAAGTAAGATTGATGGCCATATTTTGCAA TCTAATTATTGTGGCTATTTCAATTGGAGCTATCCTTTCCTCCATAGACCCTAA TAGACGCAAAGTTGATGAGACAAATGTTAATATTATATTTATAAGCCCGACA ACAGTTTCAGATTTAGCAATCGTAATCACGGCAAAACTGAACCAGTACATTT CATACTTTAGAAGAATTTTGTTATGGCAGGATTTTATTCTTTCAACCAGATTT ACTTTGTGTGTCTACATTATGGGAATCCTTTTCAAAATTATACCGCTGGTTGC TTTAATATATATAATGTGTTGGGCATTTTATCTTTATATGTTCATATGCAAGG AGATGGCAGATCAGTTGTTGGACATTATAATGGTTTATTTGAAAAGGGTGTC GGGAAATTTCGACGAGTTCTGCTGCAATATCCCCAAGATGAAGGATGTGGG TAAGGAACTG 39 ATGTTTAACGAAAACGAGATCCCTCAGTTCCAGCGCCCTTTCACCCCCTCCAA BMR1_01G02100 AGACACGGAGATTGACACTAGCATCAATTTCCCTGCCAATTCAATTCTAAATA full sequence ACATATCATTTCAAAAATTATTGGATTGGTTAGATGAATGCACAGAAGGCTT ACCAATCGTGGAAAGTTTTGCGAGACGATTCAACGTTACTAGGGGTCATATA GCGGCAATATTTTCTGCTATATTAATTCTGTACTTATTTTCGGCTGGAAAATT AACATCTTTTGCAATACAATAGGGTTGGTATATCCAGCATTTAAATCTCATAA GGTTTTACTGATTCATAGAGCCATGACTGAAAGTCCAAAAGCAACAGCTACT ATCACAACTGGGGGAAAGGACAAAGAAAATGACACAAACCCTCAAATGCCT ACTTGTCTCAATGGAATCCAAGGGGAAATTATGTTTTGGCTTAGGTACTGGA TTGTTTACTCGCTTTATTTATTTATTTCGATATTAATTTTCCCCTTGATATCATG GTTGCCACTAATATCTATAGTTCGGGTTGGTTTTATCCTATATCTCTACCATCC ATACACTAGAGGGGCCAACGCGATCTACTACCTCGTAATATCACCTCTGCTA AGCAAAAACCAGAAGATCATAGAACAGGCCATTGACACCCTCGAAAGGGTT GCGATGGGTGAAATCAGGAAATTTACAGAAAAACAAATGAAGCTACAC 40 ATGGATAGCATTGAAGAGTGTAATAAATTGGTAGATGCGGTGACCAAATTG BMR1_04G07360 GCTACATCATTTGATTACCAAGCGCAAGAATATCTATACAATCTTACCCGCGA full sequence TGAAAATGCAATTAACATTGCATTATCCTTTCTATTATCCAATAAATACGTGT TGAACTATTCAAGCAGTTACAGCAACGAAATTTTACAATATATAACCGATGTT CGTGGCCCAAGCAGTTGTTGCATCTGGGACAATATCCAATTGGACGAAAATA CCAGTAGGCATCTGGGCCACATAACGTCTATTTTGTTTAAGGAGCATATGCA TATAGTTTTGTGTTTCGACGATGCAAAGCTTATTGCACTTATTGATACATTGG TTACTATTTGGCAATTACAATCAACTATATCACTAGTAACTCAAAGCTGTTTC GCTGATAATGCTACTGATGATCATATAACGAGAGTTATTTCTATGATTATATT CAAAAAGCCGCAATTATTAACCCACTTCATCAACATACTAAACACTCCCGAG GTAACTGATAAGTATGTGGCTGATTTCTCAATGAAATTTAAATTAAGAAATT GCGATAGAGATGATATTTATGTCACAAGGTATAGGCATATGATTGTCTATGT GTTAGCTCAGGTGCTTGAAAATTTTGTGTCTTCGAACGTTACTGATAAGATAT CTTTTTTGCCAATTAACTTCTCAGATTTAGTCATATTGTCGGGAAATTGTAAC AATAGGTATCTTTACTCATGCGCGTTATTAATTGTAAATTCATTGAACTTTCA CATTTCAGAAGAGTTATTACATCAATTATTGGTAATTGTTGAGAAATCTGGAT TTGACTGCGATATACTTGAGATGTGGAATTTTGCAATCAGTCATTTACAATAT TTGAAAGTTGACAATTTCATGATGAAATCTTTCAGGTTAGTCCAAAACGGGC TCATTGAATCCACAAATCCAATAAACATTGTGCCTGTATTAAACGGTATAACA GTATACTATGTTAATAATTCATCAGTTCCAACTGAGATCGTAAGAGCTTACA GGCAATTGCTTTCCGTCGAATCCCTTGACATTCCCATTGAAATCGGTGAGTTA TTTGCAGCTGTCCGAAATGCTTGCACTGAAAGTCTGATTACGTATGATAAAA TTGCTTCATTTGTGTCGCAATTTGACATAAGAATGATTTCAAGTAGGTTAATT GAACTTTGCAATCCTAACATGCAATGTTGGTCTTGGTTGAATGTAAATTGTTG TGTAGAAGAATTTCCCACGGATTATGACATATACATTGATTCTATGAAAAAT GTATTTAAAGACCTATTTTTCATCTTACCAGAACAACTTTTAGACACTATAAT AAGTGAGATATTACAATCTGTAAATGAAAATACGGAAACGTCACTAATAGCA TTAATTTGCTATGATTGCCTAGAAGTAAGGCATGCATTGGCAATTCTAGACA AGATTAAATCATTGTTTACGCTGAAATTGTCTTCAAACTACGATTTCATCCTT ACTTGTATGTTTTGCCAATCCATATTTGTGCCTCATTTGCACAGGATAAAAGA TATATTATTCCAGAGATTTGTTATCAACATGAAAATATCGAATATTTGGTCTA AAAGACTGGCTAAATGTATAGCAATATTGAACGATAGGGATTCCGTTCATAG CGTGATTGAATGTGTTCAGTATATATTGGAAACTTCTAAATCCAAGGGTGTA TTGTCACAAAATCACTATCCTTCAATTCTATCATTGTCAAATCTTTTAGATACG TTGCCACAAATGCCTTGTGATGTTAATTTGACTGGATCCAATTATATTTTTTG CAAATGCTTGCTCGCGTTAAGTATTTCTACTAGACAATTATCACAAATATTGC CAAAAATCGCTCTCAATTTGGATATACTATATGAAGGTGATGTGGACAACGA TTTGTTTTGTTTATCAACATCTGATAGCCAAGCTGCAGTTAACTTATTGATGC AACTATATATACAAAATAAGGTAACCGATATACCTAATGAATTGCACGAGAT ACGTCACCCCGGGCTTTTAATGTTATGTAATTTGAATGAAAATTTTGAACAAT TCAGGATGAATAAGTGTGTAGAGTTCATGTCAACGGTCAGCCAAGAATCTT GCTGTCACTGTGCATGGGCCTGTATTGCCTATGCATCTTATGTTATCGAACAG AAATTCAACTTTGAATTTTCAATGGAGATGGTTAAAGTGATATATAGGGTGT TACCATGTTTAATTAAAATGGTTAAGTCCAAGAATGAGCCTTTACATTGGGA TTTAGATGCAATTTTGGAGTACAGAATTAATGAAATACATCCTTCAATTAAGT GTCTAAAGGGGACAATTTGGAATATTTACTATGATGGTACAGTTAGTGTCGG CTCTGACGCTTGTAAATATTTATCACTAGTAAATAAATATGCTCCAAGAATCA TTAAAAGTTTATCAAATCCAAATGTATTGCAGTTGGTGTATCAAATTTCAATG TGTACTGGATCCATTGAAGTTATAGCAAATAAAATCGCAATTTCACTA 41 ATGAGCTGCATATTGAAGTGTAACAACGAGGATGAATTGGTTGTGGATGGG BMR1_02G02185 GAAAAACCCAAGGTAGTAGAGTACGTCGAGAAACCATCGGTAATTTACAAG full sequence CCTACAACCGTTGTCCCCCCTAACAGCCTGATCGAAATTACTGCTCCCAAGGA TTTGCCTCAAAATCCAACATTCTTCCCAACCATAGATACATTTTTCGACAATG ACGTCAAGCAGATCGTTTTGTTGATGGAATTGCCCGGATTTGTGGCGGGAG ATATTGATTTGGAGGTTGGTGAAGGTGAAGTTTGTGTTTGCGGCCCTAGGTC TAAGGAGGAACTATATGAGAAGTATGGTCAAAATTTAGATATACACATTCGA GAACGAAAGGTTGGCTACTTTTACCGTCGCTTCAAGCTCCCGCACAATGCTC TTGACAACACCGTAAAGGCATCGTATCAAAATGGGATTCTCGAAGTGCGCAT TACCTGTACAGAATTTTCACCGAAAACGCGTGTGGAGATCACATCT 42 ATGAACGGGTCGGTGGAAGAACTATTAAATCGCTTATCGGCGATTAACGAC BMR1_04G08260 AGGTGTTATCTGCTGGTGGATGAAATAAAAAATTACATGTCAAACAAACTTT full sequence ACCATGAGTTAACCCTAGCACTCATCGAGTTGTTTACCATGTCTGAAATATCT TGCAATGACAGACTTCTTTTATTTGAAATGATAGTGCATCCTATCAAAAATGA TCTGAACATTTTGAAGTTCTCACATATTTTACGCCTGTCCTCTGAACATCTAG AGCCTTTAGCGTCACTAGATCAATTGTCTAAATATGACAATTACTTATCAACA GACACTCAAGCCAGTTTTATGATTAAAATTGCAAAGTCATACCACCATACTCG GAATCAATCCTACGACCAAAGTTTGAAGCTTTTGGAGGAAGTAAAACCTGA GATAGAATCCGGATTTGGCTTGGACATAACGGTAATCTCAGCCTATTACAAA GTGTCAGCGAATCTAAATAAGGCAACGCACAAATACAATTCATGGTACCAG GACTCACTCATGTACCTAAACTACACTCCTCTGGATAGTATCAGTCCAACTGA ACGTGATGAATTGGCACTGGATATAGCAATTGCCTCGATTGCAGCGCCAGAT AACTACAATTTTGGTGCTGTTCTAATTCAGCCACTAATCAACACGTGTTTGAA ACAGCACTCCACATTTGGCTGGGTATACGCAATTTTAATGGCTCTTAATGAT GGCGACTTCACACAATACGATGAGATAATTTCAAAATATAAAGTCCAAATCA GCCATTCCGAATTAAATCATCACAAAGAGCAACTCCAAAGAAAAATTACACT TATGGCATTCCTGAAACTCGTGTTTAGAAAGGCTAAGAAACAACGCATATTT ACATTCGAAGAAATATCCCAGAATTGTCGGATTCCAATAGATGAGGTGGAAT ACTTGCTTCTAAAAGCTATGTGCAACAATGTGGTTAAGGGTAAAATTAACCA GGTTGAGCAGATCGTCAGTTTTACGTGGGTTCAACCTAGGATAATCGACTCA ACAAAATTGACAGTTCTCCTAGATGGGGTTAACGAGTGGAATCAACAACTAA AAGCGCTCATAAACAAGCTCAAAGAAATTACTCCAGAGTTACTAGTGTCG 43 ATGTCGGGACTGTTTGGTCAATCGCAGCAGATTGGTGGTGGATTATTTGGCC BMR1_03G02345 AATCTAACCAACAGTCTGGGGGAGGGTTATTCGGGTCAACATCGCAACAGC full sequence CAACCCAAACATGCAGCGGATTATTTGGATCATCTCCAGCTCCCGCAAATAG TAGTATTTTCGGTTCAAATACACAGTCTGCTGCTTCTAGCGGTGGGATTTTTG GTTCATCCACAGCTCCAGTTAATAGCGGAGGCGGCATTTTTGGTCAATCTAA CACTAATGTTAGCAGTGGTAGTGGATTATTTGGCGGTGGAAATACTACAGG TCAATCTGGTGGGGGTATTTTTGGTTCATCAACTACTTCGGCTCCAGCATCAG GAGGAGGGTTATTCGGCCAAACTGGAACCACTACCAGTGGAGGAGGTGGA TTATTTACCTCATCTTTTGCGCCAGCGCCAAGTAGTGGCGGACTATTCGGGC AGCCATCCACACCTGCGACAAGCGGCACTGGATTGTTTTCTAGTACCAGTAC GACTCAACCAAGTTCGGGAGCGGGGCTATTTGGTTCAAGTACTACTCCAGCT AGTGGAAGCGGTGGTCTTTTTGGTCAACCTAGCACTTCAACGACAACTAGTG GTGGTATTTTTGGTTCATCAACTACTTCGGCTCCAGCATCAGGAGGAGGGTT ATTTGGCCAAACAGGAACCCCTGCCAGTGGAAGCAGTGGCATATTTGGCTC AACAAATACCACAACCTCGGCCTCAGGGACTGGATTGTTCGGTTCGACGTCA ACAACTCCACAGCCTGGCAGTGGTAGTGGATTATTTGGCGGTGGAAATACT ACAGGTCAATCTGGTGGGGGTATTTTTGGTTCATCAACTACTTCGGCTCCAG CATCAGGAGGAGGGTTATTCGGCCAAACTGGAACCACTACCAGTGGAGGTA ATTTATTTGGCACCACCAGCACTACAACACCGGCGCCTGCTAGCGGTGGACT ATTCGGTTCTTCTTCTACTACGTCCACAACAACTCCAACACAAGCAACAACTA CAGTAGGAGGGGGAGGGTTGTTTGGCACTGCAAGTGCAACTACTGCGCCG GCGTCTGGTGGATTATTTGGCACCACCAGCACCACAACACCGGCGCCTGCTA GCGGTGGACTATTCGGTTCTTCTTCTACTACGTCCACAACTCCAACACAAGCA ACAACTACAGTAGGAGGGGGAGGGTTGTTTGGCACTGCAAGTGCAACTACT GCGCCGGTGTCTGGTGGATTATTTGGCACCACCAGCACTACAACACCGGCG CCTGCTAACAACACTACTCCTGCTAACACTACAACTGTGCCAACTGCCATACT AACAACTAGTACACCGTCCCCTGCTACCGATGGTTTATTTGGATCGACCGAC GTTACAACTACTACTGATTCCACAACTAAACTCGTTGGCACTAGCCCTTTTTGA AAAACAAACAGATTCGGGGCCGGATGTCACTGCCGCTACTAACGATACGCC TAACGCATTTATCACAGATAAGCCAACAGCTGGAGGAGATCTCAAAGGCCA AGTTGAGTTGTCATTTTCGTCTGTGGAGCATGAATGTGTGCAAGACCTACTT TCTAACTGGGAGAAGAGAATGGAAGTCAAGATACAAAGGTTTACTGAGTTT GCTCAAGACATACAGAGGATCGATCGCGATCTAATACTACAAACAGAAAAG TTACAAGTATTGTTGGATGAGCAAGCAACTGTTCAGGAAAGACAGAACCAA GTACAGGAGATGATCCAAGTTATTGAGAAGGAACAGCAACAGGTGCTAGAA TCGTTGGATATAATGGATACTGCGCTTGAGACGTTGTTGGGTCCAGATAAGA AAATAACCAGTAAGAGTGGCGAAACTATAGATTTCGTCTCCAACAAATTGCG GGATTTAGAAGCTCAATTAAAGGCCGCACATGATGTGGTGGATTCTGTGGTT AAAGCTTCGCAACCGGAGCCCCTGGCTAATGTTGCTAAAGTGTTTGCATTCC ACCAGGACACTATTGAAAATATCCAGCTTCAGACATCGGAGATTGAGAAGA AACTTGACGCTATTAAGCAGCAGCAAGCAGTC 44 ATGGCATCGCTGCTGAAAGTATCTCCACAGGACAATATCGAGTTTCCACTGG BMR1_02G02960 TGTTATACACGCCTTTAAATGCGAACTTATTACTGGAAAACTTATCTGGTGTG full sequence CATGTGGCCTTTAAAATCAAGACCACAGCACCCAAGGGCTATCTAGTCCGCC CCTCAACAGGGACAATTAAACCTGGAGAGGCACTTACTGTTCAAATCATTTT GCAGCCTCTTAGTGAGGTTCCAAATGTGGTCAACGACAGGTTCCTCGTCCAG TGCACTGCCATCGCAAATGACGAATTGGTTTCCAAGGATTTCTGGACTACAC TCGACAAAGCCAGTATCCAAGATCACCGCCTCAATGTAACCTTTAAGAAGGA TATTGGTCTCAACATACAGACAAGCCAATCCAACATTGGCGTCCCGCCTCAC ATCGCCGCCAGAATTCTCACCCCCTTGGGACCCAATGCAGGCGTTGCAGAAC TAAGACAAAAGTACGAGGAGCTAGTATCATACTGTCTAACGGCAGAAAAGC AAAAGGCTGCTCTGGTTAAAGACAACGAGAAACTTCGTCAGCGCCTCCACCT GGGCCCCAACGACCCTGCCTCTGGAAACAAATGGCCCCTAGAAGGTTGGCA TCTACCCGTAATGGTGATCATATTAGTAATTATTCTCAAGGCCATTGGCTACT GG 45 ATGTCACAAGGACCTGCTATTGGGATTGATTTGGGAACCACTTACTCATGTG BMR1_01G02545 TTGGCGTTTGGAAAAATGAAACTGTAGAAATTATAGCCAATGATCAAGGCA full sequence ACCGTACAACTCCGTCTTATGTTGCCTTCACTGATGTGGAGCGATTGGTTGG CGACGCCGCCAAAAATCAGGATGCTAGAAATCCTGAAAACACAGTTTTTGAT GCAAAAAGATTGATAGGGAGAAAGATCAATGATCCCTGCATCCAAAGTGAT ATAAAACACTGGCCATTTACTGTTGCGGCTGGACCAAATGATAAGCCTGTGA TCAAGGTACAATTTCAGGGTGAAACTAAATCCTTCCACCCAGAGGAAATATC TTCTATGGTCCTCACCAAGATGAAGGAAATTGCAGAATCTTATTTGGGAAAG ACAATCTCTAATGCTGTTATCACTGTACCCGCTTATTTTAACGATTCTCAACG ACAGGCCACTAAGGACGCTGGTACCATTGCTGGGTTAAATGTTATGCGTATT ATCAATGAACCAACTGCTGCCGCAATTGCCTATGGTATGGACAAGAAAGGT ACTTCTGAAAAAAATGTGTTGATTTTCGATTTGGGCGGTGGCACTTTCGATG TATCAATTTTAACTATCGAAGATGGCATTTTTGAGGTAAAGGCCACACAAGG TGATACCCACTTGGGCGGTGAGGACTTTGACAACAGATTGGTCAACTTTTGC GTAGATGACTTTAAGCGAAAGAATGGCGGGAAGAATATTTCAACCAATAGA CGTGCATTACGTAGACTTAGAACACAATGTGAACGTGCTAAACGCACCTTAT CCCACTCAACACAGGCCACGATTGTTGTAGAAGCTATATTTGATGGCATCGA CTACAGTTGCAACATCACTAGGGCCAGATTCGAGGAGCTCTGTGCCGAAAT GTTCAAAAACACTTTAATCCCAGTCGAAAAAGCCTTAGCTGATGCAGATATG GATAAGAAGCAGATCCATGAAGTGGTACTTGTAGGAGGTTCGACCCGTATC CCTAAGATACAGCAATTGATTAAGGACTTTTTCAATGGCAAAGAACCCTGCA AATCAATTAACCCAGACGAAGCAGTAGCCTATGGCGCGGCTGTTCAGGCTG CAATTTTAACTGGCGAACAATCTAGCAAGGTCCAAGATTTACTGTTACTAGA TGTCACTCCACTGTCACTAGGACTTGAGACCGCCGGCGGTGTTATGACCGTG CTGATACCCAGAAATACTACTATCCCCGCCAAGAAGGAACAAGAATTTACAA CCAATGAAAATAACCAAACGGGGGTTATGATCCAGGTCTTCGAAGGAGAAC GTTCTATGACATGTGACAACAATTTACTAGGCAAATTCCATTTGACTGGCATA CCTCCTGCTCCAAGAGGAGTTCCCCAGATTAAAGTCACCTTTGACATCGATG CCAATGGTATATTGACAGTGTCTGCTGCCGACAAGTCGACAGGAAAGACTG AACATGTAACTATAACAAATGACAAGGGAAGACTTTCACAGCAAGACATTG ATAGGATGGTTGCTGAAGCAGAGAAGTTTAGGGAAGATGATGAAAAGAAG AAGAGGTGTGTTGAATCCAAGAATGAACTTGAGAACTATTGTTATTCAATGA AAAATGCCTTGGAAGAGGAGGGTGTTAAATCTAAATTGTCATCTTCTGAGCT TTCCGAGGCTCAGAAACTGTTACAAAACACTTTCAGTTGGATAGAATCTAAT CAATTGGCTGAGAAAGAGGAATTTGAAGCTAAACTCAAGGAGGTACAAGCT GTATGTACACCTTTGACTGCTAAATTGTACCAAGCTGGTGGTGGCGTCCCAG GTGGCGCTGCCCCAGGTGGATTTAATGCTGGCGGTGCAGCTCCCTCAGGAC CAACTGTGGAAGAAGTTGAT 46 ATGAAACTTATCGCCTGTACCCTTAAGAATGTTGAGACCTGTGTCGAAGTAG BMR1_03G04110 ACCCATCTGACACCGTAGATGCTTTAACCAATAAAATTGGATCGAGCCTGAA full sequence CAATGCTAGTGCCAGCAAAATGCGATTAATCCATGCTGGTAAAATTTTGAAA ATGGAACAAAAGATATCGGACTATTCTGACATCAAGGATGGGGATAAAATT ATTGTTCTGTTCAGCAAACAATCCGAAGCAAGTACAATAGCTAATCCTACAC CTGCCCCTACCTCTACTCCCATTGCAGATGCCAACACCAGTCCCCCGAAACCC ATCCCTACCACGGACCCTAATGCCTTACTGATGGGCGAGGAGCTAGAAAAA GCCATAAACGGTATAGTAGAAATGGGTTTTGATGTTGAATCAGTCAAAGCG GCCATGAGCGCAGCATTCAACAACCCTAACAGGGCTATAGAACTCCTCACGC GTCATGAGGTAGACGTTTCAGACCATGATACGCATCAATCTGTTCAAACGAC GGGCGTGTTGGATGAGCTGCGACAGCACCCTATGTTTGAGCAGATGCGGGC GATTGTGCGCAGTAATCCACAGACGCTGCCACAAATTTTATCGCTCATAGGG CAGTCAGACCCATCATTACTCCAAGCCATCACTGAAAATCAGGAAGAATTTA TCCAGCTACTGAGTGAACCGGTACTAGGCACAAGTGGTGATTTTATTGACGC CCAGTCTATAACTTTGACACCAGAGGAAATGGAGTCTATAAACAGACTGGA GGGGCTCGGGTTTTCGAGACCGGCAGC1GICGAGGCATTTTTGGCATGCGA TAAAAATGAGGAGATGGCAGCAAATTATTTGCTTGAAAACATAGCAGATTAT GTCTCTGACAATGATAAT 47 ATGGATGGACAAACGGAACAACAATTGATTGAGGAAAATATCGAAAGATTT BMR1_02G02560 AGGATACTTTATCAATTGCATTTGGACAACAATGAACCCTCTCCCACCGCTCA full sequence ATTTAATTATGCATGTGCTCTAGTCTGCTCCAATCAGCGATCACATAATGACA CAGCTATATATCTCTTAGATGAATTGGTTCGCATTAGATATGAAAGTGAAGA ATGTTTTATCAACTCGCTCTTGCACATATGAAACGCAGAAGTTTCGTCAAGT CAAAGGAATACTTGGATCGTATCATTGCACTGGAGGGTTCTAATCAACGTGT AATGGCACTTAAATCAGTAGTCGTTTCATTACTAGCTCAGGATACTTTCATGG GCGGATTACTTGGCGCCACTGCAGCGTTCGCAATAATTCTCTTCTTCACAATG AAAAGGAATACC 48 ATGAACCAGAATTCACTCTGTTGTTCCAAGGGTTTTACCATGTTTGCAGGTG BMR1_04G05635 GTGCCTTTTTCCTAGTTTCATTTAAGCCGGAAACTGTTATAACGAATCCACTG full sequence CTGTTACGGCCGCTGTTGAATGTGAGCTGGGGGTATATATTTGGATCCCATC TATGGGCAGCAATATCTACCTATAACAAAAAATATTGGGAAAACCGGATAAT TCCTGATTATGCTAATTCACCATCACGGGAGATTATGCAAAGCAGACTTAAG ATTAACGAATCGCGCATTTATCGTATATACCTAGAGAATCTAATCCAAACAA ACGTTTTAGCGAATGGGATATTATTAGTTACAACAAGCGCATTGGCGCCTTC AAACAAATTTCTAAGGATTTGCTCGGGAGCAGCACTGCTTTTATCCATAGGA AATGTAGTATTTGCGTTGCCTGAAGACGAAAAGGATGATGATGTGGGAGTT GTAAAGACATCTTCTACTTCATGTTTCCTGTCTGAAGTACTCTCTTTTTGCACA TTCGGCGCCATAGTCCCTTATGTATTTGCA

    [0142] Anti-Babesia microti Antibody-Based Compositions

    [0143] Also provided herein are compositions that include one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, or 24 or more) antibodies or antigen-binding fragments thereof which specifically bind to one or more Bm antigens or antigenic fragments thereof. These compositions optionally include a pharmaceutically acceptable excipient, preservative, carrier or diluent.

    [0144] In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 1, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 2, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 3, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 4, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 5, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 6, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 7, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 8, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of SEQ ID NO: 9, or one or more antigenic fragments thereof.

    [0145] In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof, and to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof. In some embodiments, the one or more antibodies bind to one or more Bm antigens that each comprise an amino acid sequence having at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 95%, 97%, 99%, or greater) to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0146] Antibodies and antigen-binding fragments, variants, or derivatives thereof include, but are not limited to, monoclonal, genetically engineered, and otherwise modified forms of antibodies, including, for example, chimeric antibodies, humanized antibodies, primatized antibodies, heteroconjugate antibodies (e.g., bi-, tri-, and quadri-specific antibodies), and antigen-binding fragments of antibodies including Fab, Fab′, F(ab′)2, Fv, scFv, tandem scFv, diabody, triabody, tetrabody, small modular immunopharmaceutical (SMIP), nanobody or other single domain antibody. Also included are affibodies, isolated CDRs, and a combination of two or more isolated CDRs.

    [0147] Antibodies can be generated using any of the numerous methods for making antibodies known in the art. Monoclonal antibodies that specifically bind to one or more Bm antigens can be prepared using standard hybridoma technology (see, e.g., Kohler et al., Nature 256:495-7, 1975; Kohler et al., Eur. J. Immunol. 6:511-9, 1976; Kohler et al., Eur. J. Immunol. 6:292-5, 1976; Hammerling et al., Monoclonal Antibodies and T Cell Hybridomas, Elsevier, N Y, 1981). Antigen-binding fragments can be produced by recombinant DNA techniques, enzymatic or chemical cleavage of intact antibodies, or, in certain cases, by chemical peptide synthesis procedures known in the art. Antigen-binding fragments also can be identified by screening a phage display library (Vaughan et al., Nat. Biotechnol. 14:309-14, 1996) using a polypeptide of the invention described above. See, for example, the references cited herein above, as well as Zhiqiang An (Editor), Therapeutic Monoclonal Antibodies: From Bench to Clinic. 1st Edition. Wiley 2009, and also Greenfield (Ed.), Antibodies: A Laboratory Manual. (Second edition) Cold Spring Harbor Laboratory Press 2013, for methods of making recombinant antibodies, including antibody engineering, use of degenerate oligonucleotides, 5′-RACE, phage display, and mutagenesis; antibody testing and characterization; antibody pharmacokinetics and pharmacodynamics; antibody purification and storage; and screening and labeling techniques.

    [0148] Useful antibodies can be identified using one of several screening assays, including ELISA, immunoprecipitation or Western blot analysis. In one example, antibodies are assayed by ELISA to determine whether they are specific for the immunizing antigen (e.g., one or more Bm polypeptides as described herein). Using standard techniques, some wells of a plate are coated with the immunogen whereas other wells are coated with irrelevant Bm antigens. An aliquot of the antibody sample is added to each well of the plate. The unbound material is washed, and the bound antibody detected by use of a second antibody specific for the immunoglobulin of the species in which the antibody was generated. Antigen-binding fragments can be screened for utility in the same manner as intact antibodies.

    [0149] In some embodiments, the anti-Bm antibodies used in the therapeutic or prophylactic methods of the invention can be purified from cells known to produce monoclonal antibodies, e.g., hybridomas as understood by those of skill in the art. In other embodiments, the anti-Bm antibodies used in the therapeutic or prophylactic methods of the invention can be artificially manufactured molecules, e.g., recombinant antibodies. In some embodiments, a therapeutic or prophylactic antibody may comprise monoclonal antibodies along with other components such as excipient, diluent, or carrier (e.g., human serum albumin). A therapeutic or prophylactic antibody may further comprise preservatives or combinations thereof, as will be readily understood by those of skill in the art.

    [0150] Pharmaceutical Formulations

    [0151] The Bm antigens, the Bm nucleic acid molecules, and the Bm-specific antibodies described herein can be formulated into pharmaceutical compositions that are biologically compatible and suited for administration to a subject

    [0152] Compositions may be administered to a subject in a variety of forms which depend on the route of administration, as will be understood by those skilled in the art. The compositions described herein may be administered by the oral route or a parenteral route, including, for example, the intravenous, intraperitoneal, subcutaneous, intradermal, intramuscular, and topical routes.

    [0153] A composition described herein may be orally administered, for example, with an inert diluent or an edible carrier. The composition may be enclosed in hard- or soft-shell gelatin capsules, or compressed into tablets, or incorporated directly with the food of the diet. A composition described herein may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers. Compositions suitable for buccal or sublingual absorption include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerin. Nucleic acid-based compositions can optionally be formulated in a drug delivery vehicle such as, e.g., a lipid nanoparticle or a dendritic cell, in the case of an RNA vaccine. DNA vaccines, e.g., plasmid vaccines, can be formulated in saline or sucrose solutions, as is known in the art.

    [0154] A composition described herein may also be administered parenterally. Solutions of a composition described herein can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2012, 22nd ed.) and in The United States Pharmacopeia: The National Formulary (USP 41 NF 36), published in 2018. The pharmaceutical forms suitable for injectable use include dispersions or aqueous solutions, and powders for the extemporaneous preparation of injectable dispersions or aqueous solutions. In all cases, the forms must be sterile and must be fluid to the extent that they may be easily administered via syringe. Compositions for injectable use may be administered by continuous infusion, single or multiple boluses, or through the use of microneedles. Compositions suitable for topical administration include lotions, creams, ointments, gels, foam, transdermal patches, pastes, and tinctures. Nucleic acid-based compositions, such as DNA vaccines, can be administered by injection (e.g., intramuscular or intradermal injection). DNA vaccine administration can optionally include the use of a gene gun and, optionally, gold or tungsten microparticles onto which the DNA has been absorbed.

    [0155] The compositions described herein may be administered to a subject, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the composition, the route of administration, and standard pharmaceutical practice.

    [0156] Methods

    [0157] The invention provides methods for determining whether a subject has natural protective immunity against Bm; immunizing a subject against Bm; determining whether a prophylactic regimen has conferred protective immunity against Bm; identifying a subject who is likely to benefit from an anti-Bm antibody based therapy; treating a subject having babesiosis with an anti-Bm antibody based therapy; and optimizing administration of an anti-Bm antibody regimen. The methods described herein are based on the findings that a) resolution of B. microti infection in cd4-deficient mice requires antibody-producing B cells and antibody class switching, b) such resolution is concomitant with the accumulation of B. microti-specific IgG antibodies in blood, and c) these IgG antibodies target a restricted set of B. microti polypeptides.

    [0158] Methods for Monitoring Immune Protection

    [0159] The invention provides methods for detecting particular combinations of a subset of Bm-specific antibodies that can serve as indicators of protection against Bm, including protection acquired naturally during the course of an infection with Bm, protection induced by a Bm antigen-containing composition, a Bm antigen-encoding composition, or an anti-Bm antibody composition so as to achieve prophylaxis against Bm, and protection induced by an anti-Bm antibody composition so as to treat a subject experiencing babesiosis.

    [0160] The invention provides methods for determining whether a subject has protective immunity against Bm. In certain instances, the methods include the use of immunoassays. In one example, the methods include applying a body fluid sample from a subject to a solid support containing one or more Bm antigens or antigenic fragments thereof (e.g., as described herein); applying an antibody detection agent to the solid support; and determining whether the fluid sample contains IgG antibodies that are specifically reactive to the one or more Bm antigens or antigenic fragments thereof. In certain embodiments, the methods further include identifying the subject as likely to have protective immunity against Bm if the sample from this subject is determined to have a sufficient titer of one or more IgG antibodies that bind to one or more Bm antigens or antigenic fragments thereof, that is, a titer in the fluid sample that is above a cutoff titer.

    [0161] The invention also provides methods for identifying a patient experiencing babesiosis as likely to benefit from an anti-Bm antibody-based therapy. In certain instances, the methods include the use of immunoassays. In one example, the methods include applying a body fluid sample from a subject to a solid support containing one or more Bm antigens or antigenic fragments thereof (e.g., as described herein); applying an antibody detection agent to the solid support; and determining whether the fluid sample contains antibodies that are specifically reactive to the one or more Bm antigens or antigenic fragments thereof. In certain embodiments, the methods further include identifying the subject as likely to benefit from an anti-Bm antibody based therapy if the sample from the subject is determined to have an insufficient titer of one or more IgG antibodies specific for one or more Bm antigens or antigenic fragments thereof, that is, a titer in the fluid sample that is below a cutoff titer. In some embodiments, the methods further include administering to the subject an effective amount of a composition comprising one or more anti-Bm antibodies if the subject has been identified as likely to benefit from administration of the one or more anti-Bm antibodies.

    [0162] The invention further provides methods for optimizing the administration of an anti-Bm antibody-based therapy to a subject experiencing babesiosis. In certain instances, the methods include the use of immunoassays. In one example, the methods include applying a body fluid sample from a subject to a solid support containing one or more Bm antigens or antigenic fragments thereof (e.g., as described herein); applying an antibody detection agent to the solid support; and determining whether the fluid sample contains antibodies that are specifically reactive to the one or more Bm antigens or antigenic fragments thereof. In certain embodiments, the methods further include administering to the subject an effective amount of a composition comprising one or more anti-Bm antibodies if the sample from the subject is determined to have an insufficient titer of one or more IgG antibodies specific for one or more Bm antigens or antigenic fragments thereof, that is, a titer in the fluid sample that is below a cutoff titer.

    [0163] Any of the compositions described herein may be used in conjunction with any of the above-described methods. In certain embodiments, the one or more Bm antigens or antigenic fragments thereof (e.g., as described herein) are provided as an array affixed to a solid phase. In other embodiments, antibody reactivity is determined by ELISA, western blot analysis, or rapid diagnostic tests such as a lateral flow assay, or an assay employing a microfluidic device.

    [0164] In some embodiments, the body fluid is a blood sample or a cell-free fraction thereof (e.g., a serum or plasma sample). In certain embodiments, the subject from whom body fluid is acquired was previously immunized with a Bm antigen-containing composition, a Bm antigen-encoding composition, or an anti-Bm antibody composition (e.g., a composition described herein). In certain embodiments, the detection of certain Bm-specific antibodies in body fluids of previously immunized individuals is used to ascertain immune protection against Bm. In other embodiments, the subject from whom body fluid is acquired was previously treated with an anti-Bm antibody composition (e.g., a composition described herein). In some embodiments, the detection of certain Bm-specific antibodies in body fluids of individuals previously treated with therapeutic antibodies is used to ascertain immune protection against babesiosis.

    [0165] Methods for Conferring Prophylaxis

    [0166] The invention provides methods for immunizing or conferring protective immunity against Bm in a subject who does not experience babesiosis. In some embodiments, the methods include administering to the subject a composition including one or more Bm antigens or antigenic fragments thereof (e.g., as described herein). In some embodiments, the methods include administering to the subject a composition including one or more nucleic acid molecules (e.g., DNA or RNA) encoding one or more Bm antigens or antigenic fragments (e.g., as described herein). In other embodiments, the methods include administering to the subject a composition including one or more antibodies or antigen-binding fragments thereof that specifically bind to one or more Bm antigens or antigenic fragments thereof (e.g., as described herein).

    [0167] In some instances, the subject has been determined to be at risk of Bm infection or risk of experiencing babesiosis. In certain instances, the subject has been determined to have a titer of IgG antibodies specific for one or more Bm antigens (e.g., as described herein) that is below a cutoff titer in accordance with the diagnostic methods described herein. In some embodiments, administration of a composition as described herein for prophylaxis against Bm reduces duration and/or severity of a future episode of babesiosis.

    [0168] Methods for Treating Babesiosis

    [0169] The invention provides methods for treating a subject who experiences babesiosis (e.g., mild or severe, including persistent or relapsing babesiosis). In some embodiments, the methods include administering to the subject an effective amount of a composition comprising one or more anti-Bm antibodies or antigen-binding fragments thereof that are specific for one or more Bm antigens (e.g., as described herein).

    [0170] The invention also provides methods for supplementing an immune response against Bm in a subject who experiences babesiosis (e.g., mild or severe, including persistent or relapsing babesiosis). In some embodiments, the methods include administering to the subject an effective amount of a composition comprising one or more anti-Bm antibodies or antigen-binding fragments thereof that are specific for one or more Bm antigens (e.g., as described herein). In certain embodiments, prior to administration, the subject has been determined to have insufficient titers of IgG antibodies specific for one or more Bm antigens, that is, titers that are below cutoff titers for these Bm antigens.

    [0171] In some embodiments of any of the above-described methods, treatment reduces duration and/or severity of symptoms of babesiosis. In other embodiments, treatment reduces duration of infection with the etiologic agent of babesiosis. In some embodiments, the etiologic agent of babesiosis is Babesia microti.

    [0172] Selection of Subjects

    [0173] Subjects who may benefit from the methods described herein are subjects who experience or are at risk of experiencing babesiosis (e.g., mild or severe babesiosis, including persistent or relapsing babesiosis). Patients at risk of experiencing mild babesiosis include otherwise healthy individuals of age 40 years and above. Patients at risk of experiencing severe babesiosis include those who lack a spleen; those who suffer from anatomical or functional hyposplenism; those who are or were recently treated with rituximab or any other antibody that depletes mature B cells, including those diagnosed with a B cell lymphoma or an autoimmune disease; those for whom the CD4 T cell compartment has been depleted, including those experiencing HIV/AIDS or treated with an immunosuppressive therapy for stem cell or solid organ transplantation; and those who are immunosuppressed by treatment of comorbidity, including those who suffer from a malignancy and those who are treated from a chronic inflammatory disorder. Patients at risk of developing mild or severe babesiosis are individuals who are exposed to ticks and those who are transfused with blood products, particularly packed red blood cells.

    [0174] Subjects who may be treated using the methods described herein include those who received a diagnosis of babesiosis that was made following the standard and routine diagnostic procedures known in the art. An exemplary diagnostic procedure is the microscopic evaluation of thick or thin blood smears after exposure to the Giemsa stain or the Wright stain. Another exemplary diagnostic procedure is the amplification of Babesia genomic DNA by use of the polymerase chain reaction and of Babesia specific primers. In certain embodiments, a subject has already undergone treatment for babesiosis but such treatment has not been sufficient to achieve cure. In other embodiments, the subject has yet to be treated for babesiosis.

    [0175] Doses and Dosages

    [0176] A composition of the invention is administered to a subject (e.g., a mammal, such as a human or mouse) in an effective amount, which is an amount that produces a desirable effect in the treated subject. Effective and optimal doses and dosages for the compositions of the invention can be determined using methods known in the art. Single or multiple administrations of the compositions of the invention can be carried out to attain the desirable effect. Doses and dosages can be selected by the treating physician. It is anticipated that optimal doses and dosages will vary with the age, weight, and health of the subject. It is also anticipated that optimal doses and dosages will vary with the mode of administration, that is, a lower amount of the composition will be needed to attain the desirable effect when administered by the intravenous route as compared with other parental routes such as the intradermal, subcutaneous and intramuscular routes.

    [0177] Guidance to identify effective and optimized doses and dosages of the composition is provided in the exemplified approach described herein. For example, in the context of prophylaxis, subjects are immunized with varying doses at varying intervals, and are evaluated for the titers of antibodies that are specific for one or more Bm antigens. Immunized subjects also are evaluated for their protection from babesiosis caused by a subsequent challenge with a Babesia species. Such results can be used to optimize doses and dosages required for effective immunization of subjects, e.g., humans, mice and other mammals. Results can be extrapolated by persons having skill in the art.

    [0178] In the context of prophylaxis, compositions of the invention are administered to a subject (e.g., a human or mouse) in an amount sufficient to delay, reduce, or preferably prevent the onset of the disorder (e.g., babesiosis). By way of example, a composition comprising one or more Bm antigens or antigenic fragments thereof, or one or more nucleic acid molecules encoding one or more Bm antigens or antigenic fragments thereof, may be administered to a subject at a dose in a range of 1 μg/kg to 1,000 μg/kg (e.g., 1 to 1,000 μg/kg, 5 to 1,000 μg/kg, 10 to 1,000 μg/kg, 1 to 750 μg/kg, 5 to 750 μg/kg, 10 to 750 μg/kg, 1 to 500 μg/kg, 5 to 500 μg/kg, 10 to 500 μg/kg, 1 to 100 μg/kg, 5 to 100 μg/kg, 10 to 100 μg/kg, 1 to 50 μg/kg, 5 to 50 μg/kg, or 10 to 50 μg/kg).

    [0179] In therapeutic applications, compositions of the invention are administered to a subject (e.g., a human or mouse) already suffering from babesiosis in an amount sufficient to achieve cure, reduce the severity of one or more symptoms associated with babesiosis, or prevent or reduce the complications of babesiosis. By way of example, a composition comprising one or more anti-Bm antibodies or antigen-binding fragments thereof may be administered to a subject at a dose in a range of 50 mg/kg to 2,000 mg/kg (e.g., 50 to 2,000 mg/kg, 100 to 1,500 mg/kg, 200 to 1,000 mg/kg, 300 to 750 mg/kg, or 400 to 500 mg/kg).

    [0180] Combination Treatments

    [0181] A pharmaceutical composition including one or more anti-Bm antibodies or antigen-binding fragments thereof that specifically bind one or more Bm antigens as described herein, can be administered alone or in combination with one or more therapeutic agents. For example, a pharmaceutical composition including one or more anti-Bm antibodies or antigen-binding fragments thereof that specifically bind one or more Bm antigens as described herein can be administered in combination with standard-of-care antibiotics used in the treatment of babesiosis. Antibiotic regimens used to treat babesiosis include two-drug regimens (e.g., atovaquone and azithromycin; atovaquone and clindamycin; or clindamycin and quinine), three-drug regimens (e.g., atovaquone, azithromycin, and clindamycin; or atovaquone, proguanil, and azithromycin), four-drug regimens (e.g., atovaquone, azithromycin, clindamycin, and quinine; or atovaquone, proguanil, azithromycin, and clindamycin). In combination treatments, one or more of the therapeutic agents may be administered at a lower dose or dosage than the standard dose and dosage used when administered alone. If combined, therapeutic agents should be administered at doses and dosages that provide a therapeutic effect. Doses and dosages may be determined empirically from combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005).

    [0182] Evaluation of Efficacy

    [0183] The efficacy of the methods and compositions of the invention in preventing or treating babesiosis can be readily ascertained by those of skill in the art by means of (a) evaluating clinical manifestations associated with Babesia infection, including elevated temperature, chills, sweats, anorexia, headache, or (b) detecting abnormal laboratory parameters associated with Babesia infection, including low red blood counts, low platelet counts, elevated liver enzymes, or (c) monitoring the presence of Babesia parasites by microscopic evaluation of peripheral blood smears or PCR-based amplification of Babesia DNA. Thus, according to the methods of the present invention, a subject shows little to no clinical manifestations and/or few to no laboratory parameter abnormalities when titers of IgG antibodies specific for one or more Bm antigens are above cutoff titers for these Bm antigens.

    Kits

    [0184] The invention also provides kits or articles of manufacture containing materials useful for the prevention of babesiosis, treatment of babesiosis, and/or monitoring of individuals undergoing prophylactic or therapeutic administration of the compositions of the invention.

    [0185] Any of the compositions described herein can be provided in a kit for use in accordance with any of the prophylactic and therapeutic methods described herein. By way of example, a kit of the invention may include one or more Bm antigens or antigenic fragments thereof, as described herein. By way of another example, a kit of the invention may include one or more nucleic acid molecules (e.g., DNA or RNA) that encode one or more Bm antigens or an antigenic fragment thereof. By way of another example, a kit of the invention may include one or more antibodies or antigen-binding fragments thereof that are specific for one or more Bm antigens, as described herein. These kits can include a package insert that instructs a user of the kit, such as a physician, to perform the methods described herein. These kits may optionally include a syringe and a needle or another device for administering the composition.

    [0186] Any of the composition comprising of one or more Bm antigens can be provided in a kit for use in accordance with the methods described herein to monitor individuals undergoing prophylactic or therapeutic administration of any one of the compositions of the invention. In one example, such a kit includes (a) one or more Bm antigens or antigenic fragments thereof, as described herein, optionally immobilized on one or more solid supports; (b) an antibody detection reagent; and (c) a package insert comprising instructions for using the one or more Bm antigens and the antibody detection reagent in accordance with any of the methods described herein.

    EXAMPLES

    [0187] The following examples illustrate certain aspects of the invention and are not to be considered as limiting the scope thereof.

    Example 1: Host IgG Antibodies are Required for Resolution of Bm Infection in Cd4-Deficient Mice

    [0188] Mice expressing the cd4 gene (“WT mice”) or lacking the cd4 gene (“cd4-deficient mice” or “cd4−/−mice”) were infected with Bm and monitored over time for Bm parasitemia, i.e., the frequency of red blood cells infected with Bm (FIG. 1). Cd4-deficient mice experienced higher peak parasitemia than WT mice but mice from both strains resolved the infection. Depletion of B cells, antibody-producing cells, by administration of 18612, a monoclonal antibody directed against mouse CD20, did not alter the inherent resistance of WT mice but prevented the resolution of Bm infection in cd4-deficient mice (FIG. 1). Administration of 2B8, an irrelevant monoclonal antibody that recognizes human CD20 but not mouse CD20, failed to alter the course of parasitemia in WT mice and in cd4-deficient mice (FIG. 1). The importance of B cells to the resolution of Bm parasitemia in cd4-deficient mice was confirmed by the persistence of Bm parasitemia in cd4-deficient mice that lacked the igh6 gene (FIG. 2).

    [0189] In cd4-deficient mice, resolution of Bm parasitemia was concomitant with the accumulation in blood of Bm-specific IgG antibodies, but not of Bm-specific IgM antibodies (FIG. 2). To test the hypothesis that IgG antibodies are critical for resolution of parasitemia, cd4-deficient mice that lack the aicda gene were generated. Lack of AICDA, the enzyme required for antibody class switching, including to the IgG class, prevented full resolution of Bm parasitemia in cd4-deficient mice (FIG. 3). Overall, these results strongly suggest that IgG antibodies are required for full resolution of Bm parasitemia in cd4-deficient mice.

    [0190] To assess whether IgG antibodies protect from persistent Bm parasitemia by engaging Fc receptors, cd4-deficient mice that lack fcer1 g, the gene which encodes the chain common to all activating Fcγ receptors, or fcgr2b, the inhibitory Fcγ receptor, were generated. Lack of either receptor did not modify the resolution of Bm parasitemia in cd4-deficient mice (FIG. 4), indicating that the Fc portion of IgG antibodies is dispensable for resolution of Bm parasitemia. To test whether IgG antibodies protect from persistent Bm parasitemia by activating the complement system, cd4-deficient mice that lack c3, the gene that encodes the complement component C3, were generated. Lack of c3 delayed but did not prevent full resolution of Bm parasitemia in cd4-deficient mice (FIG. 5), indicating that the complement component C3 and the downstream effectors of the complement system which include C5 and the terminal membrane attack complex are not required for resolution of Bm parasitemia in cd4-deficient mice. Moreover, Bm parasitemia in C57BL/6 mice that lacked c3 was as low as that in wild-type C57BL/6 mice, indicating that neither the complement component C3 nor its downstream effectors are critical for host resistance to Bm. Taken together, these observations indicate that antibody-mediated neutralization of Bm antigens can protect a host from persistent Bm infection even when the Fc portion of the antibodies is removed or when the antibodies are designed to leave the complement system intact.

    [0191] To characterize the IgG repertoire produced by cd4-deficient mice following Bm infection, peripheral blood was collected at times of peak parasitemia, partial resolution, and full resolution (FIG. 6). Peripheral blood was also collected from wild-type mice infected with Bm (FIG. 6). For each blood sample, plasma was separated and probed for IgG reactivity to Bm antigens using microarray chips, as described in Example 2, below.

    Example 2: A Few Bm Proteins are Targeted by Host IgG Antibodies in Cd4-Deficient Mice

    [0192] Proteome Microarray Chip Fabrication and Design

    [0193] A large number of Bm genes (˜91% coverage of the LabS1 strain genome) were cloned into the expression vector pXT7. Included were the genes corresponding to SEQ ID NOs: 1-24. Custom polymerase chain reaction (PCR) primers comprising 20-bp gene-specific sequences tagged with 33-bp adapter sequences were used to amplify Bm genomic DNA. These adapter sequences, which flank the target amplicons, were homologous to adapter sequences found at the ends of the linearized T7 expression vector pXT7. Such homology allowed amplicons to be cloned by in vivo homologous recombination in competent DH5α cells. The clones were verified by amplifying the inserted genes using sequence-specific PCR primers and sequencing the resulting amplicons. For protein microarray chip fabrication, Bm proteins were expressed in an E. coli-based cell-free in vitro transcription and translation (IVTT) system (RTS 100 E. Coli HY Kit from BiotechRabbit, Berlin, Germany) according to manufacturer's instructions. Expressed Bm proteins were printed onto nitrocellulose-coated glass AVID slides (Grace Bio-Labs, Inc., Bend, Oreg.) using an OmniGrid Accent microarray printer (DigiLab, Inc., Marlborough, Mass.). Fabricated proteins microarray chips were QC'ed using monoclonal anti-polyhistidine (clone His-1; Sigma-Aldrich, St. Louis, Mo.) and anti-hemagglutinin (clone 3F10; Roche, Indianapolis, Ind.) antibodies.

    [0194] Each chip was spotted with 4,333 peptide fragments representing proteins from 3,184 unique Bm genes as well as 176 IgG positive control spots and 61 spotted IVTT reactions without Bm ORFs (IVTT controls). The IgG positive control spots served as an assay control, while the IVTT control spots served as a sample-level normalization factor. For each chip, 3 replicates were printed on 3 nitrocellulose “pads.”

    [0195] Plasma Probing

    [0196] Plasma samples were diluted 1:100 in an E. coli lysate solution (3 mg/mL) in protein arraying buffer (Maine Manufacturing, Sanford, Me., USA) and incubated at room temperature for 30 min. Chips were rehydrated in blocking buffer for 30 min. Blocking buffer was removed, and chips were exposed to diluted plasma samples using sealed, fitted slide chambers to avoid cross-contamination between pads. Chips were incubated overnight at 4° C. with agitation. Chips were washed five times with TBS-0.05% Tween 20, and exposed at room temperature to biotin-conjugated goat anti-mouse IgM and Cy3-conjugated goat anti-mouse IgG (Jackson ImmunoResearch, West Grove, Pa., USA) diluted 1:500 and 1:200, respectively, in blocking buffer. Chips were washed three times with TBS-0.05% Tween 20 and exposed at room temperature to streptavidin-conjugated SureLight P-3 (Columbia Biosciences, Frederick, Md., USA) in the dark. Chips were washed three times with TBS-0.05% Tween 20, three times with TBS, and once with water. Chips were air-dried by centrifugation at 1,000×g for 4 min and scanned on a GenePix 4300A High-Resolution microarray scanner (Molecular Devices). Spot and background intensities were measured using an annotated grid file (.GAL).

    [0197] Protein Microarray Data Analysis

    [0198] Raw spot and local background fluorescence intensities, spot annotations and sample phenotypes were imported and merged in the R statistical environment, where all subsequent procedures were performed (www.r-project.org). Foreground spot intensities were adjusted for local background by subtraction, and negative values were converted to 1. All foreground values were transformed using the base 2 logarithm (log 2). The dataset was normalized to remove systematic effects by subtracting the median signal intensity of the IVTT controls for each sample. Given that the IVTT control spots carry the chip, sample and batch-level systematic effects, but also antibody background activity to the IVTT system, this procedure normalizes the data and provides a relative measure of specific antibody binding to non-specific antibody binding (a.k.a. background). For the normalized data, a value of 0.0 indicates that the intensity does not differ from background whereas a value of 1.0 (log 2) denotes an intensity that is twice that of background. Immunoreactive antigens were defined as those for which a mean immunoreactivity of at least 1.0 log 2 (i.e., at least twice that of background) was detected at time of peak infection, time of partial resolution, or time of full resolution of infection.

    [0199] Results

    [0200] In cd4-deficient mice, as parasitemia resolves, the number of IgG reactive Bm antigens markedly increased (FIG. 7A). Sixteen were IgG reactive at time of peak infection, 35 at time of partial resolution, and 52 at time of full resolution (FIG. 7B).

    [0201] In WT mice, at time of full resolution, the IgG antibody repertoire was restricted to a set of 60 Bm antigens (FIG. 8, top panel, left of the vertical dashed line). Of these 60 antigens, only 41 were IgG reactive when plasma was obtained from cd4-deficient mice (FIG. 8, middle panel, left of the vertical dashed line and above the horizontal dashed line). Eleven additional antigens which were not reactive to plasma obtained from WT mice were reactive, although marginally, to plasma obtained from cd4-deficient mice (FIG. 8, middle panel, right of the vertical dashed line). Thus, cd4 deficiency results in the loss of IgG reactivity to 19 Bm antigens but in the gain of IgG reactivity to 11 Bm antigens. Such shift in the repertoire of cognate Bm antigens can be attributed to the fact that absence of CD4 curtails the T cell response to low-affinity antigens while promoting the T cell response to high-affinity antigens.

    Example 3: Characterization of Bm Antigens Associated with Protective Immunity

    [0202] Sixteen distinct antigens were identified at time of peak infection as exhibiting an IgG reactivity that was above background by greater than 2.0-fold (>1.0 log 2) (FIG. 9). Neutralization of these Bm antigens by IgG antibodies may help curtail the rise in Bm parasitemia. Their gene ID and their name (if unnamed, a brief description of their characteristics) are provided in FIG. 10.

    [0203] Sixteen antigens were identified as exhibiting a significant gain in IgG reactivity of greater than 2.0-fold (>1.0 log 2) from time of peak infection to time of partial resolution (FIGS. 11+12). Neutralization of these Bm antigens may help achieve partial resolution of Bm parasitemia. Their gene ID and their name (if unnamed, a brief description of their characteristics) are provided in FIG. 13. Of these 16 antigens, only seven were not identified as immunoreactive at time of peak infection (see hatched bars in FIG. 11; numbers highlighted by a hexagon in FIG. 12; antigen names written in black ink in FIG. 13).

    [0204] Ten antigens were identified as exhibiting a significant gain in IgG reactivity of greater than 2.0-fold (>1.0 log 2) from time of partial resolution to time of full resolution (FIGS. 14+15). Neutralization of these Bm antigens may help achieve full resolution of Bm parasitemia. Their gene ID and their name (if unnamed, a brief description of their characteristics) are provided in FIG. 16. Of these 10 antigens, only one was not identified as immunoreactive at time of peak infection or as gaining reactivity from time of peak infection to time of partial resolution (see dotted bar in FIG. 14; number highlighted by a rectangle in FIG. 15; antigen name written in black ink in FIG. 16).

    [0205] To characterize the 24 distinct antigens identified as exhibiting IgG reactivity at time of peak infection and/or gaining IgG reactivity from one time point to the next, we analyzed their amino acid sequence for the presence of a signal peptide. We also tested for an inverse relationship between IgG reactivity and Bm parasitemia at times of peak infection and/or partial resolution. Of the 24 antigens, nine exhibit a motif that can act as a signal peptide (FIG. 19). Of the eight antigens that contain a signal peptide and were immunoreactive at time of peak infection and/or had gained reactivity by the time of partial resolution, three displayed an immunoreactivity that was inversely correlated with Bm parasitemia at one of these two time points (FIGS. 17+18). These three antigens, namely BMR1_01 G03280 (SEQ ID NO: 1), BMR1_04 G05532 (SEQ ID NO: 2), and BMR1_01 G00985 (SEQ ID NO: 3), are referred to as group #1 antigens (FIGS. 19+20).

    [0206] Of the eight antigens that contain a signal peptide and were immunoreactive at time of peak infection and/or had gained significant immunoreactivity by the time of partial resolution, five displayed an immunoreactivity that was not inversely correlated with Bm parasitemia at one of these two time points. These five antigens, namely BMR1_02 G01760 (SEQ ID NO: 4), BMR1_03 G00365 (SEQ ID NO: 5), BMR1_03 G04695 (SEQ ID NO: 6), BMR1_03 G03430 (SEQ ID NO: 7) and BMR1_04 G06070 (SEQ ID NO: 8), are referred to as group #2 antigens. This group of antigens also includes BMR1_04 G09385 (SEQ ID NO: 9), an antigen which contains a signal peptide but for which a significant gain in IgG reactivity was only detected at time of full resolution, thereby precluding the testing of a relationship between IgG reactivity and Bm parasitemia at this time point.

    [0207] A third group of antigens comprise the 15 antigens that do not contain a signal peptide but were identified as immunoreactive at time of peak infection or as gaining immunoreactivity from one time point to the next. The gene ID of each of these 15 antigens is provided in FIG. 20. One of these 15 antigens, namely BMR1_04 G07360 (SEQ ID NO: 16), displayed an immunoreactivity that was inversely correlated with Bm parasitemia at time of partial resolution of infection (FIG. 18).

    [0208] Any of the 24 distinct antigens, which are listed in FIG. 20, may be used individually or in combination as a Bm antigen vaccine to confer protection from babesiosis, or may be targeted by a therapeutic antibody composition to resolve or help resolve babesiosis.

    Example 4: RNA Vaccines

    [0209] RNA molecules encoding one or more of the Bm antigens described herein (or one or more antigenic fragments thereof) are generated using standard methods (e.g., in vitro transcription of DNA templates) and formulated for administration to subjects. Optionally, modified nucleosides are used in the synthesis of the RNA molecules to reduce immunogenicity and/or to increase stability. RNA molecules are administered to subjects, e.g., by injection of lipid nanoparticles comprising the RNA.

    Example 5: DNA Vaccines

    [0210] DNA vaccines can be prepared using expression vectors that are engineered to include sequences encoding one or more of the Bm antigens described herein (or one or more antigenic fragments thereof). In addition to the antigen coding sequence(s), the vectors can include promoter sequences (for example, a viral promoter, e.g., a Rous Sarcoma Virus (RSV) promoter, a cytomegalovirus (CMV) immediate early promoter, or an SV40 promoter), a polyadenylation/transcription termination signal, and optionally other standard vector sequences, as are known in the art. The vectors can be mono- or poly-cistronic, as is known in the art. DNA vaccines can be administered to subjects by injection (e.g., intramuscular or intradermal injection), gene gun, or mucosal delivery.

    Other Embodiments

    [0211] Various modifications and variations of the described invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the art are intended to be within the scope of the invention.

    [0212] Some embodiments are within the scope of the following numbered paragraphs.

    [0213] 1. A composition comprising (a) one or more Babesia microti (Bm) antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof; and (b) a pharmaceutically acceptable carrier or diluent.

    [0214] 2. The composition of paragraph 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    [0215] 3. The composition of paragraph 2, further comprising one or more Bm antigens each comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    [0216] 4. The composition of paragraph 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    [0217] 5. The composition of any one of paragraphs 1-4, wherein the composition comprises two or more Bm antigens or antigenic fragments thereof.

    [0218] 6. The composition of paragraph 5, wherein the composition comprises three or more Bm antigens or antigenic fragments thereof.

    [0219] 7. The composition of paragraph 6, wherein the composition comprises four, five, six, seven, eight, or more Bm antigens or antigenic fragments thereof.

    [0220] 8. The composition of paragraph 7, wherein the composition comprises nine or more Bm antigens or antigenic fragments thereof.

    [0221] 9. The composition of paragraph 8, wherein the composition comprises twenty-four Bm antigens or antigenic fragments thereof.

    [0222] 10. The composition of paragraph 1, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0223] 11. The composition of paragraph 2, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    [0224] 12. The composition of paragraph 3, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    [0225] 13. The composition of paragraph 4, wherein the one or more Bm antigens each comprise an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    [0226] 14. The composition of paragraph 1 or 10, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-24.

    [0227] 15. The composition of paragraph 2 or 11, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-3.

    [0228] 16. The composition of paragraph 3 or 12, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 4-9.

    [0229] 17. The composition of paragraph 4 or 13, wherein the one or more Bm antigens each comprise the amino acid sequence of any one of SEQ ID NOs: 1-9.

    [0230] 18. A composition comprising (a) one or more nucleic acid molecules encoding one or more Bm antigens, or one or more antigenic fragments thereof, and optionally (b) a pharmaceutically acceptable carrier or diluent, wherein the one or more Bm antigens, or the one or more antigenic fragments thereof, are as specified in any one of paragraphs 1-17.

    [0231] 19. The composition of paragraph 18, wherein the one or more nucleic acid molecules comprise an RNA molecule or a DNA molecule, which optionally comprises a sequence of any one or more of SEQ ID NOs: 25-48, a fragment thereof, or a codon-optimized version thereof.

    [0232] 20. The composition of any one of paragraphs 1-19, further comprising an adjuvant.

    [0233] 21. The composition of any one of paragraphs 1-20, wherein the composition is formulated for administration by the oral route or a parenteral route, such as a parenteral route selected from the group consisting of the intravenous, intraperitoneal, subcutaneous, intramuscular, and topical routes.

    [0234] 22. A composition comprising one or more antibodies that each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof.

    [0235] 23. The composition of paragraph 22, wherein the one or more antibodies each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-3, or one or more antigenic fragments thereof.

    [0236] 24. The composition of paragraph 23, further comprising one or more antibodies that each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-9, or one or more antigenic fragments thereof.

    [0237] 25. The composition of paragraph 22, wherein the one or more antibodies each specifically bind to one or more Bm antigens that each comprise an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-9, or one or more antigenic fragments thereof.

    [0238] 26. The composition of any one of paragraphs 22-25, further comprising a pharmaceutically acceptable carrier or diluent.

    [0239] 27. A method of immunizing or conferring protective immunity against babesiosis to a subject, the method comprising administering the composition of any one of paragraphs 1-21 to the subject.

    [0240] 28. A method of immunizing or conferring protective immunity against babesiosis to a subject, the method comprising administering the composition of any one of paragraphs 22-26 to the subject.

    [0241] 29. The method of paragraph 27 or 28, wherein the subject does not experience babesiosis.

    [0242] 30. The method of paragraph 29, wherein treatment reduces the parasite burden upon a subsequent challenge.

    [0243] 31. The method of paragraph 29 or 30, wherein treatment reduces the severity of babesiosis upon a subsequent challenge.

    [0244] 32. The method of paragraph 28, wherein the subject experiences babesiosis.

    [0245] 33. The method of paragraph 32, wherein treatment reduces the parasite burden.

    [0246] 34. The method of paragraph 32 or 33, wherein treatment reduces the severity of babesiosis.

    [0247] 35. The method of any one of paragraphs 27-34, wherein the subject experiences mild or severe babesiosis, including persistent or relapsing babesiosis.

    [0248] 36. A method of supplementing an immune response in a subject experiencing babesiosis, the method comprising administering to the subject the composition of any one of paragraphs 22-26.

    [0249] 37. A method of treating babesiosis in a subject in need thereof, the method comprising administering to the subject the composition of any one of paragraphs 22-26.

    [0250] 38. The method of any one of paragraphs 27-37, wherein the etiology of babesiosis is Babesia microti.

    [0251] 39. A method for determining whether a subject, prior to administration of a composition of any one of paragraphs 1-21 and paragraphs 22-26, has protective immunity against Bm-induced babesiosis, the method comprising:

    [0252] a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof;

    [0253] b. applying an antibody detection agent to the solid support of (a); and

    [0254] c. identifying the subject as likely to have protective immunity against Bm-induced babesiosis if the fluid sample from the subject is determined to have a sufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof of, that is, a titer in the fluid sample above a cutoff titer for the one or more Bm antigens.

    [0255] 40. A method for determining whether a subject, following administration of a composition of any one of paragraphs 1-21 and paragraphs 22-26, has acquired protective immunity against Bm-induced babesiosis, the method comprising:

    [0256] a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof;

    [0257] b. applying an antibody detection agent to the solid support of (a); and

    [0258] c. identifying the subject as likely to have protective immunity against Bm-induced babesiosis if the fluid sample from the subject is determined to have a sufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof of, that is, a titer in the fluid sample above a cutoff titer for the one or more Bm antigens.

    [0259] 41. A method for identifying a patient who experiences babesiosis and is likely to benefit from an anti-Bm antibody-based therapy, the method comprising:

    [0260] a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof;

    [0261] b. applying an antibody detection agent to the solid support of (a); and

    [0262] c. identifying the patient as likely to benefit from an anti-Bm antibody based therapy if the fluid sample from the subject is determined to have an insufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof, that is, a titer in the fluid sample below a cutoff titer for the one or more Bm antigens.

    [0263] 42. A method of optimizing the administration or therapeutic efficacy of an anti-Bm antibody-based therapy to a subject experiencing babesiosis, the method comprising:

    [0264] a. applying a body fluid sample from the subject to a solid support, wherein the solid support comprise one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof;

    [0265] b. applying an antibody detection agent to the solid support of (a); and

    [0266] c. administering to the subject the composition of any one of paragraphs 22-26 if the sample from the subject is determined to have an insufficient titer of one or more IgG antibodies that specifically react with one or more Bm antigens of (a) or antigenic fragments thereof, that is, a titer below a cutoff titer for the one or more Bm antigens.

    [0267] 43. The method of paragraph 41 or 42, further comprising administering to the subject the composition of any one of paragraphs 22-26, wherein the subject has been determined as likely to benefit from an anti-Bm antibody based therapy or from a modified dose or dosage of anti-Bm antibody based therapy.

    [0268] 44. The method of any one of paragraphs 27-43, wherein the subject is a human.

    [0269] 45. A kit comprising (a) one or more Bm antigens comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-24, or one or more antigenic fragments thereof, wherein one or more Bm antigens are immobilized on one or more solid supports; (b) an antibody detection reagent; and (c) a package insert comprising instructions for using the one or more Bm antigens and the antibody detection reagent in accordance with the methods of any one of paragraphs 40-43.

    [0270] 46. A kit comprising (a) the composition of any one of paragraphs 1-21 and (b) a package insert comprising instructions for using the composition in accordance with the methods of any one of paragraphs 27, 29, and 44.

    [0271] 47. A kit comprising (a) the composition of any one of paragraphs 22-26 and (b) a package insert comprising instructions for using the composition in accordance with the methods of any one of paragraphs 28, 32, and 44.

    [0272] Other embodiments are within the scope of the claims.