Btnl9 And Ermap As Novel Inhibitors Of The Immune System For Immunotherapies

Abstract

Provided are methods of treating a tumor in a subject with a BTNL9-binding antibody. Also provided are methods of treating a tumor in a subject with an ERMAP-binding antibody. A fusion protein comprising a BTNL9 or ERMAP and related compositions and encoding nucleic acids are also provided.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of U.S. Provisional Application No. 62/084,124, filed Nov. 25, 2014, the contents of which are hereby incorporated by reference.

BACKGROUND OF THE INVENTION

[0003] The disclosures of all publications, patents, patent application publications and books referred to herein, are hereby incorporated by reference in their entirety into the subject application to more fully describe the art to which the subject invention pertains.

[0004] Cancers, autoimmune diseases, infectious diseases, and transplantation rejection are serious public health problems in the US and other countries. The immune system, particularly T cells, plays critical roles in these diseases.

[0005] The B7 ligand family and CD28 receptor family control T cell activation and function. Both CTLA-4 and PD-1 are members of the CD28 family, and an antibody against CTLA-4 (Yervoy from Bristol-Myers Squibb) and an antibody against PD-1 (Keytruda, Merck) were approved by the FDA as new drugs for melanoma in 2011 and 2014, respectively. PD-L1 is a member of the B7 family, and antibodies to PD-L1 are in clinical trials with cancer patients. In additional, CTLA-4-Ig (Orencia) was approved by the FDA as a new drug for adult rheumatoid arthritis.

[0006] The existing technologies work by blockade of the B7/CD28 family members. The butyrophilin family is related to the B7 family, but their expression and functions in the immune system are largely unknown.

[0007] The present invention provides addresses the need for improved therapies and therapeutics based on targeting BTNL9 or ERMAP.

SUMMARY OF THE INVENTION

[0008] A method of treating a tumor in a subject comprising administering to the subject an amount of a BTNL9-binding antibody, or BTNL9-binding fragment thereof, sufficient to inhibit a BTNL9 and treat the tumor.

[0009] A method of treating a tumor in a subject comprising administering to the subject an amount of a ERMAP-binding antibody, or ERMAP-binding fragment thereof, sufficient to inhibit a ERMAP and treat the tumor.

[0010] A method of treating a tumor in a subject comprising administering to the subject an amount of a BTNL2-binding antibody, or BTNL2-binding fragment thereof, sufficient to inhibit a BTNL2 and treat the tumor.

[0011] A method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble BTNL9 to treat the autoimmune disease.

[0012] A method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble ERMAP to treat the autoimmune disease.

[0013] A method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble BTNL2 to treat the autoimmune disease.

[0014] An isolated, recombinant fusion polypeptide comprising a BTNL9 fused to an immunoglobulin polypeptide.

[0015] An isolated, recombinant fusion polypeptide comprising an ERMAP fused to an immunoglobulin polypeptide.

[0016] An isolated, recombinant fusion polypeptide comprising a BTNL2 fused to an immunoglobulin polypeptide.

[0017] An isolated chimeric nucleic acid encoding an isolated recombinant fusion polypeptide as described herein.

[0018] A composition comprising an isolated recombinant fusion polypeptide as described herein and a carrier.

BRIEF DESCRIPTION OF THE DRAWINGS

[0019] FIG. 1A-1B. mRNA transcripts of BTN and BTNL family members in mouse tissues and antigen-presenting cells relative to the reference gene OAZ1. (A) Transcripts in naive mouse tissues. Transcripts of BTN family members (upper panel) show that BTN transcripts are most abundant in mammary tissue. Transcripts of BTNL family members with intracellular SPRY domains (center panel) and of those without intracellular SPRY domains (bottom panel) are highest in the small and large intestines. Transcripts are also generally detectable in other tissues, particularly the primary lymphoid organs; MOG transcripts are highest in brain tissue. SP, spleen; LN, pooled inguinal and brachial lymph nodes; TH, thymus; BM, bone marrow; MLN, mesenteric lymph node; BR, brain; LU, lung; HE, heart; LI, liver; ST, stomach; SM, small intestine; LG, large intestine; PA, pancreas; KI, kidney; BL, bladder; SV, seminal vesicles; PR, prostate; EP, epididymis; TE, testis; FT, Fallopian tube; VA, vagina; UT, uterus; OV, ovary; MA, mammary gland. (B) Transcripts in resting and activated antigen-presenting cells (APCs). Transcripts for most genes are detectable in all three professional APCs, are in most cases lower in macrophages compared to dendritic cells and B cells, and are generally not strongly up- or down-regulated upon activation by LPS or PMA+ionomycin.

[0020] FIG. 2A-2C. Most recombinant BTN and BTNL family members inhibit the function of anti-CD3-activated primary mouse T cells. T cells were isolated from mouse spleen and lymph nodes by magnetic cell separation and activated with 2.5 .mu.g/mL plate-bound anti-CD3 in the presence of 4 .mu.g/mL plate-bound BTN- and BTNL-human Ig recombinant fusion proteins. (A) All BTN and BTNL fusion proteins, with the exception of ERMAP and MOG, reduce the metabolic activity of CD4+ T cells by 3 days post-activation. Human Ig (hIg), B7x-hIg, and B7.2-hIg fusion proteins are included as baseline, negative, and positive controls for activation, respectively. (B) BTN and BTNL fusion proteins reduce proliferation of activated CD4+ and CD8+ T cells. CD90.2+ T-cells were pulsed with CFSE prior to activation for 4 days, and stained with fluorescence-labeled antibodies to CD4 and CD8. CD4+ T cells (upper panel) showed reduced proliferation in the presence of BTN, BTN2, BTNL1, BTNL2 (full- and partial length), BTNL4, BTNL6, BTNL9, and ERMAP, and CD8+ T cells (panel) showed reduced proliferation in the presence of BTN, BTN2, BTNL1, BTNL2 (full- and partial-length), BTNL4, BTNL6, and BTNL9. The division indices, i.e., average number of divisions (closed bars), and percent divided (open bars) of CD4+ and CD8+ T cells are shown to the right of the histograms. (C) Secretion of the cytokines IFN-.gamma., IL-2, TNF-.alpha., and IL-17A by CD4+ T cells 2 days post-activation is reduced in the presence of BTN and BTNL fusion proteins compared to baseline (hIg).

[0021] FIG. 3A-3B. CD4+ (A) and CD8+ (B) T cells activated for 3 days express a receptor for BTN and BTNL proteins. Total lymph node cells were activated by plate-bound 2.5 .mu.g/mL anti-CD3 and incubated with biotinylated BTN-, BTNL-hIg, or hIg-fusion protein, stained with fluorescence-labeled antibodies to CD4 and CD8, and evaluated for fusion protein binding with fluorescence-labeled streptavidin. Both activated T cell subsets express a receptor for all BTN and BTNL family members, with the exception of MOG, which is consistent with functional data.

[0022] FIG. 4. The IgV1 domain of human BTNL2 bound human CD8 T cells, human B cells, and human NK cells. BTNL2-IgV1-Ig protein (open histograms) and control Ig (shaded histograms) are shown in FACS assays.

[0023] FIG. 5. The IgV1 domain of human BTNL9 bound human CD8 T cells, human B cells, and human NK cells. BTNL9-IgV1-Ig protein (open histograms) and control Ig (shaded histograms) are shown in FACS assays.

DETAILED DESCRIPTION OF THE INVENTION

[0024] This invention provides the first disclosure that butyrophilin-like 9 (BTNL9) and erythroblast membrane-associated protein (ERMAP), two members of the butyrophilin family, inhibit T cell functions. A short form of BTNL2 was also strongly inhibitory. In particular, evidence that recombinant proteins of BTNL9 and ERMAP inhibit cell proliferation and cytokine production of T cells and that activated T cells have receptors for BTNL9 and ERMAP is provided. Since BTNL9 and ERMAP are expressed by immune cells or other cells. Applications of targeting these two molecules to enhance immunity (e.g. cancer immunotherapy) or decrease immunity (e.g. therapy of autoimmune diseases) are encompassed. Also, applications of using BTNL2, especially the short or "partial" form are encompassed.

[0025] BTNL9 is an inhibitor of the immune system. Therefore blockade of BTNL9-mediated immune suppression with blockers (e.g. monoclonal antibodies to BTNL9) can be used for treatment of human cancers and infectious diseases, while enhancement of BTNL9-mediated immune suppression with soluble proteins (e.g. BTNL9-Ig) can be used for treatment of autoimmune diseases and transplantation.

[0026] ERMAP is an inhibitor of the immune system. Therefore blockade of ERMAP-mediated immune suppression with blockers (e.g. monoclonal antibodies to ERMAP) can be used for treatment of human cancers and infectious diseases, and enhancement of ERMAP-mediated immune suppression with soluble proteins (e.g. ERMAP-Ig) can be used for treatment of autoimmune diseases and transplantation.

[0027] A method of treating a tumor is provided in a subject comprising administering to the subject an amount of a BTNL9-binding antibody, or BTNL9-binding fragment thereof, sufficient to inhibit a BTNL9 and treat the tumor.

[0028] Also provided is a method of treating a tumor in a subject comprising administering to the subject an amount of a ERMAP-binding antibody, or ERMAP-binding fragment thereof, sufficient to inhibit a ERMAP and treat the tumor.

[0029] Also provided is a method of treating a tumor in a subject comprising administering to the subject an amount of a BTNL2-binding antibody, or BTNL2-binding fragment thereof, sufficient to inhibit a BTNL2 and treat the tumor.

[0030] In an embodiment, the BTNL9 is a human BTNL9. In an embodiment, the ERMAP is a human ERMAP. In an embodiment, the BTNL2 is a human BTNL2.

[0031] In an embodiment of the methods, the tumor is a tumor of a breast, lung, thyroid, melanoma, pancreas, ovary, liver, bladder, colon, prostate, kidney, esophagus, or is a hematological tumor, or wherein the tumor is a lymphoid organ tumor.

[0032] In an embodiment of the methods, the antibody is administered as an adjunct to an additional anti-cancer therapy for the tumor.

[0033] In an embodiment of the methods, the amount of a BTNL9-binding antibody is administered. In an embodiment of the methods, the BTNL9-binding antibody binds an IgV1 domain of human BTNL9.

[0034] In an embodiment of the methods, the amount of a ERMAP-binding antibody is administered.

[0035] In an embodiment of the methods, the amount of a BTNL2-binding antibody is administered. In an embodiment of the methods, the BTNL2-binding antibody binds an IgV1 domain of human BTNL2.

[0036] In an embodiment of the methods, the fragment of the antibody is administered.

[0037] In an embodiment of the methods, the antibody is a monoclonal antibody.

[0038] In an embodiment of the invention, the BTNL9 is human BTNL9. In an embodiment human BTNL9 protein has the sequence:

TABLE-US-00001 (SEQ ID NO: 1) MVDLSVSPDSLKPVSLTSSLVFLMHLLLLQPGEPSSEVKVLGPEYPIL ALVGEEVEFPCHLWPQLDAQQMEIRWFRSQTFNVVHLYQEQQELPGRQ MPAFRNRTKLVKDDIAYGSVVLQLHSIIPSDKGTYGCRFHSDNFSGEA LWELEVAGLGSDPHLSLEGFKEGGIQLRLRSSGWYPKPKVQWRDHQGQ CLPPEFEAIVWDAQDLFSLETSVVVRAGALSNVSVSIQNLLLSQKKEL VVQIADVFVPGASAWKSAFVATLPLLLVLAALALGVLRKQRRSREKLR KQAEKRQEKLTAELEKLQTELDWRRAEGQAEWRAAQKYAVDVTLDPAS AHPSLEVSEDGKSVSSRGAPPGPAPGHPQRFSEQTCALSLERFSAGRH YWEVHVGRRSRWFLGACLAAVPRAGPARLSPAAGYWVLGLWNGCEYFV LAPHRVALTLRVPPRRLGVFLDYEAGELSFFNVSDGSHIFTFHDTFSG ALCAYFRPRAHDGGEHPDPLTICPLPVRGTGVPEENDSDTWLQPYEPA DPALDWW.

[0039] In an embodiment of the invention, the ERMAP is human ERMAP. In an embodiment human ERMAP protein has the sequence:

TABLE-US-00002 (SEQ ID NO: 2) MEMASSAGSWLSGCLIPLVFLRLSVHVSGHAGDAGKFHVALLGGTAEL LCPLSLWPGTVPKEVRWLRSPFPQRSQAVHIFRDGKDQDEDLMPEYKG RTVLVRDAQEGSVTLQILDVRLEDQGSYRCLIQVGNLSKEDTVILQVA APSVGSLSPSAVALAVILPVLVLLIMVCLCLIWKQRRAKEKLLYEHVT EVDNLLSDHAKEKGKLHKAVKKLRSELKLKRAAANSGWRRARLHFVAV TLDPDTAHPKLILSEDQRCVRLGDRRQPVPDNPQRFDFVVSILGSEYF TTGCHYWEVYVGDKTKWILGVCSESVSRKGKVTASPANGHWLLRQSRG NEYEALTSPQTSFRLKEPPRCVGIFLDYEAGVISFYNVTNKSHIFTFT HNFSGPLRPFFEPCLHDGGKNTAPLVICSELHKSEESIVPRPEGKGHA NGDVSLKVNSSLLPPKAPELKDIILSLPPDLGPALQELKAPSF.

[0040] In an embodiment of the invention, the BTNL2 is human BTNL2. In an embodiment human ERMAP protein has the sequence:

TABLE-US-00003 (SEQ ID NO: 3) MVDCPRYSLSGVAASFLFVLLTIKHPDDFRVVGPNLPILAKVGEDALL TCQLLPKRTTAHMEVRWYRSDPAMPVIMYRDGAVVTGLPMEGYGGRAE WMEDSTEEGSVALKIRQVQPSDDGQYWCRFQEGDYWRETSVLLQVAAL GSSPNIHVEGLGEGEVQLVCTSRGWFPEPEVHWEGIWGEKLMSFSENH VPGEDGLFYVEDTLMVRNDSVETISCFIYSHGLRETQEATIALSERLQ TELVSVSVIGHSQPSPVQVGENIELTCHLSPQTDAQNLEVRWLRSRYY PAVHVYANGTHVAGEQMVEYKGRTSLVTDAIHEGKLTLQIHNARTSDE GQYRCLFGKDGVYQEARVDVQVTAVGSTPRITREVLKDGGMQLRCTSD GWFPRPHVQWRDRDGKTMPSFSEAFQQGSQELFQVETLLLVTNGSMVN VTCSISLPLGQEKTARFPLSDSKI.

[0041] In an embodiment of the invention, the short form of BTNL2 is a short form of human BTNL2. In an embodiment the short form of human BTNL2 has the sequence:

TABLE-US-00004 (SEQ ID NO: 4) MVDCPRYSLSGVAASFLFVLLTIKHPDDFRVVGPNLPILAKVGEDALL TCQLLPKRTTAHMEVRWYRSDPAMPVIMYRDGAVVTGLPMEGYGGRAE WMEDSTEEGSVALKIRQVQPSDDGQYWCRFQEGDYWRETSVLLQVAAL GSSPNIHVEGLGEGEVQLVCTSRGWFPEPEVHWEGIWGEKLMSFSENH VPGEDGLFYVEDTLMVRNDSVETISCFIYSHGLRETQEATIALSERLQ TELVSVSVIGHSQPSPVQVG.

[0042] Protein sequence of the IgV1 domain of human BTNL2:

TABLE-US-00005 (SEQ ID NO: 5) KQSEDFRVIGPAHPILAGVGEDALLTCQLLPKRTTMHVEVRWYRSEPS TPVFVHRDGVEVTEMQMEEYRGWVEWIENGIAKGNVALKIHNIQPSDN GQYWCHFQDGNYCGETSLLLKVAGLGSAPSIHM.

[0043] Protein sequence of the IgV1 domain of human BTNL9:

TABLE-US-00006 (SEQ ID NO: 6) EVKVLGPEYPILALVGEEVEFPCHLWPQLDAQQMEIRWFRSQTFNVVH LYQEQQELPGRQMPAFRNRTKLVKDDIAYGSVVLQLHSIIPSDKGTYG CRFHSDNFSGEALWELEVAGLGSDPHLS.

[0044] In an embodiment of the methods, the tumor is a tumor of a breast, lung, thyroid, melanoma, pancreas, ovary, liver, bladder, colon, prostate, kidney, esophagus, or is a hematological tumor. In an embodiment of the methods, the tumor is a hematological tumor and is a leukemia or a lymphoma. In an embodiment of the methods, the tumor is a tumor of the breast and is a triple negative breast cancer. In an embodiment of the methods, the tumor is a tumor of a lymphoid organ.

[0045] Also provided is an isolated fusion protein comprising a soluble portion of an BTNL9 and an Fc portion of an immunoglobulin G.

[0046] Also provided is an isolated fusion protein comprising a soluble portion of an ERMAP and an Fc portion of an immunoglobulin G.

[0047] An isolated chimeric nucleic acid encoding an isolated fusion protein as described herein is provided.

[0048] A composition comprising the isolated fusion protein as described herein and a carrier is provided.

[0049] In an embodiment, the composition is a pharmaceutical composition, and the carrier is a pharmaceutical carrier.

[0050] Also provided is a method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble BTNL9 to treat the autoimmune disease.

[0051] Also provided is a method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble ERMAP to treat the autoimmune disease.

[0052] Also provided is a method of treating an autoimmune disease in a subject comprising administering to the subject an amount of an isolated, plasma-soluble BTNL2 to treat the autoimmune disease.

[0053] In an embodiment, the BTNL-2 comprises SEQ ID NO:4 but does not comprise SEQ ID NO:3.

[0054] In an embodiment, the plasma-soluble BTNL9, or the plasma-soluble ERMAP, respectively, comprises an BTNL9 fused to an immunoglobulin polypeptide, or an ERMAP fused to an immunoglobulin polypeptide, respectively.

[0055] In an embodiment, the plasma-soluble BTNL2 comprises an BTNL2 fused to an immunoglobulin polypeptide.

[0056] In an embodiment, the immunoglobulin polypeptide comprises an Fc portion of an immunoglobulin G.

[0057] Also provided is an isolated, recombinant fusion polypeptide comprising a BTNL9 fused to an immunoglobulin polypeptide. Also provided is an isolated, recombinant fusion polypeptide comprising an IgV1 domain of human BTNL9 fused to an immunoglobulin polypeptide. In an embodiment, the IgV1 domain of human BTNL9 comprises SEQ ID NO:6.

[0058] Also provided is an isolated, recombinant fusion polypeptide comprising an ERMAP fused to an immunoglobulin polypeptide.

[0059] Also provided is an isolated, recombinant fusion polypeptide comprising a BTNL2 fused to an immunoglobulin polypeptide. In an embodiment, the BTNL-2 comprises SEQ ID NO:4 but does not comprise SEQ ID NO:3. Also provided is an isolated, recombinant fusion polypeptide comprising an IgV1 domain of human BTNL2 fused to an immunoglobulin polypeptide. In an embodiment, the IgV1 domain of human BTNL2 comprises SEQ ID NO:5.

[0060] Also provided is an isolated chimeric nucleic acid encoding an isolated recombinant fusion polypeptide as described herein.

[0061] Also provided is a composition comprising the isolated recombinant fusion polypeptide as described herein and a carrier. In an embodiment, the compositions is a pharmaceutical composition, and comprises a pharmaceutical carrier.

[0062] The term "ERMAP-Ig" fusion protein as used herein means a fusion protein constructed of a portion of an immunoglobulin and an active portion of a ERMAP, or proteins having an identical sequence thereto. In a preferred embodiment, the active portion of a ERMAP is a soluble portion of ERMAP. In an embodiment, the ERMAP has the sequence of a human ERMAP. In an embodiment, the portion of an immunoglobulin is a portion of an IgG or an IgM. In an embodiment, it as a portion of an IgG. The IgG portion of the fusion protein can be, e.g., any of an IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgG4 or a portion thereof. In an embodiment, the portion is an Fc region. In an embodiment the fusion protein comprises a sequence identical to an Fc portion of a human IgG1, human IgG2, human IgG2a, human IgG2b, human IgG3 or human IgG4. In an embodiment the fusion protein comprises a sequence identical to an Fc portion of a human IgG1. The term "Fc region" herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine of the Fc region may be removed, for example, by recombinantly engineering the nucleic acid encoding the fusion protein.

[0063] The term "BTNL9-Ig" fusion protein as used herein means a fusion protein constructed of a portion of an immunoglobulin and an active portion of a BTNL9, or proteins having an identical sequence thereto. In an embodiment, the active portion of a BTNL9 is a soluble portion of BTNL9. In an embodiment, the BTNL9 has the sequence of a human BTNL9.

[0064] The term "BTNL2-Ig" fusion protein as used herein means a fusion protein constructed of a portion of an immunoglobulin and an active portion of a BTNL2, or proteins having an identical sequence thereto. In an embodiment, the active portion of a BTNL2 is a soluble portion of BTNL2. In an embodiment, the BTNL2 has the sequence of a human BTNL2. In an embodiment, the BTNL-2 comprises SEQ ID NO:4 but does not comprise SEQ ID NO:3.

[0065] In an embodiment, the portion of an immunoglobulin of the fusion proteins is a portion of an IgG or an IgM. In an embodiment, it as a portion of an IgG. The IgG portion of the fusion protein can be, e.g., any of an IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgG4 or a portion thereof. In an embodiment, the portion is an Fc region. In an embodiment the fusion protein comprises a sequence identical to an Fc portion of a human IgG1, human IgG2, human IgG2a, human IgG2b, human IgG3 or human IgG4. In an embodiment the fusion protein comprises a sequence identical to an Fc portion of a human IgG1. The term "Fc region" herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine of the Fc region may be removed, for example, by recombinantly engineering the nucleic acid encoding the fusion protein.

[0066] In an embodiment, the Fc portion of the Ig is used in the fusion proteins as described herein. The presence of the Fc domain markedly increases the plasma half-life of the attached protein, which prolongs therapeutic activity. In addition, the Fc domain also enables the fusion protein to interact with Fc-receptors. In an embodiment, the ERMAP-Ig comprises a ERMAP portion linked to an Fc domain. In an embodiment, the ERMAP portion is bound directly by a peptide bond to the Fc domain. In an embodiment, the ERMAP portion is linked to the Fc domain through a linker. In an embodiment, the BTNL9-Ig comprises a BTNL9 portion linked to an Fc domain. In an embodiment, the BTNL9 portion is bound directly by a peptide bond to the Fc domain. In an embodiment, the BTNL9 portion is linked to the Fc domain through a linker.

[0067] In an embodiment, the fusion protein (or fusion polypeptide) is linked via a peptide linker which permits flexibility. In an embodiment, the linker is rigid. In an embodiment the linker is cleavable. Non-limiting examples of flexible linkers within the scope of the invention are G.sub.n, and GGGGS, and (GGGGS).sub.n where n=2, 3, 4 or 5 (SEQ ID NO:7). Non-limiting examples of rigid linkers within the scope of the invention are (EAAAK).sub.n (SEQ ID NO:8), (XP).sub.n. Non-limiting examples of cleavable linkers within the scope of the invention include disulfide links and protease cleavable linkers. In a preferred embodiment, the linker is a peptide linker.

[0068] In an embodiment, the Fc domain has the same sequence or 95% or greater sequence similarity with a human IgG1 Fc domain. In an embodiment, the Fc domain has the same sequence or 95% or greater sequence similarity with a human IgG2 Fc domain. In an embodiment, the Fc domain has the same sequence or 95% or greater sequence similarity with a human IgG3 Fc domain. In an embodiment, the Fc domain has the same sequence or 95% or greater sequence similarity with a human IgG4 Fc domain. In an embodiment, the Fc domain is not mutated. In an embodiment, the Fc domain is mutated at the CH2-CH3 domain interface to increase the affinity of IgG for FcRn at acidic but not neutral pH (Dall'Acqua et al, 2006; Yeung et al, 2009).

[0069] In an embodiment, the fusion protein described herein is recombinantly produced. In an embodiment, the fusion protein is produced in a eukaryotic expression system. In an embodiment, the fusion protein produced in the eukaryotic expression system comprises glycosylation at a residue on the Fc portion corresponding to Asn297.

[0070] In an embodiment, the fusion protein is a homodimer. In an embodiment, the fusion protein is monomeric. In an embodiment, the fusion protein is polymeric.

[0071] In an embodiment, a BTNL9-Ig is prepared by fusing the coding region of the extracellular domain of a BTNL9 having the same sequence as a human extracellular domain of a BTNL9 to a polypeptide having the same sequence as a human IgG1 Fc. Such can be made in any way known in the art, including by transfecting an appropriate cell type with a recombinant nucleic acid encoding the fusion protein. The BTNL2-Ig fusion protein can be made in an analogous matter, as can the other fusion proteins mentioned herein.

[0072] In an embodiment, a ERMAP-Ig is prepared by fusing the coding region of the extracellular domain of a ERMAP having the same sequence as a human extracellular domain of a ERMAP to a polypeptide having the same sequence as a human IgG1 Fc. Such can be made in any way known in the art, including by transfecting an appropriate cell type with a recombinant nucleic acid encoding the fusion protein.

[0073] In an embodiment, of all aspects of the invention described herein reciting a subject, the subject is a human

[0074] Cancers, including tumors, treatable by the invention include of the nasopharynx, pharynx, lung, bone, brain, sialaden, stomach, esophagus, testes, ovary, uterus, endometrium, liver, small intestine, appendix, colon, rectum, gall bladder, pancreas, kidney, urinary bladder, breast, cervix, vagina, vulva, prostate, thyroid, skin, or is a glioma. In an embodiment, the cancer treated is a metastatic melanoma.

[0075] This invention also provides a composition comprising a fusion protein as described herein. In an embodiment, the composition is a pharmaceutical composition. In an embodiment the composition or pharmaceutical composition comprising one or more of the fusion proteins described herein is substantially pure with regard to the fusion protein. A composition or pharmaceutical composition comprising one or more of the fusion proteins described herein is "substantially pure" with regard to the antibody or fragment when at least about 60 to 75% of a sample of the composition or pharmaceutical composition exhibits a single species of the fusion protein. A substantially pure composition or pharmaceutical composition comprising one or more of the fusion proteins described herein can comprise, in the portion thereof which is the fusion protein, 60%, 70%, 80% or 90% of the fusion protein of the single species, more usually about 95%, and preferably over 99%. Fusion protein purity or homogeneity may be tested by a number of means well known in the art, such as polyacrylamide gel electrophoresis or HPLC.

[0076] The invention also encompasses compositions comprising the described fusion proteins and a carrier. The carrier may comprise one or more pharmaceutically-acceptable carrier components. Such pharmaceutically-acceptable carrier components are widely known in the art.

[0077] In an embodiment, the subject being treated for cancer via a method herein is also treated with a chemotherapeutic agents, such as a cytotoxic agent. In an embodiment, the cytotoxic agent is doxorubicin. In an embodiment, the cytotoxic agent is a maytansinoid. In an embodiment, the cytotoxic agent an alkylating agent, an anti-metabolite, a plant alkaloid or terpenoid, or a cytotoxic antibiotic. In embodiments, the cytotoxic agent is cyclophosphamide, bleomycin, etoposide, platinum agent (cisplatin), fluorouracil, vincristine, methotrexate, taxol, epirubicin, leucovorin (folinic acid), or irinotecan.

[0078] Administration as used herein, unless otherwise stated, can be auricular, buccal, conjunctival, cutaneous, subcutaneous, endocervical, endosinusial, endotracheal, enteral, epidural, via hemodialysis, interstitial, intrabdominal, intraamniotic, intra-arterial, intra-articular, intrabiliary, intrabronchial, intrabursal, intracardiac, intracartilaginous, intracaudal, intracavernous, intracavitary, intracerebral, intracisternal, intracorneal, intracoronary, intradermal, intradiscal, intraductal, intraepidermal, intraesophagus, intragastric, intravaginal, intragingival, intraileal, intraluminal, intralesional, intralymphatic, intramedullary, intrameningeal, intramuscular, intraocular, intraovarian, intraepicardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrasinal, intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intratubular, intratumor, intratympanic, intrauterine, intravascular, intravenous, intraventricular, intravesical, intravitreal, laryngeal, nasal, nasogastric, ophthalmic, oral, oropharyngeal, parenteral, percutaneous, periarticular, peridural, rectal, inhalationally, retrobulbar, subarachnoid, subconjuctival, sublingual, submucosal, topically, transdermal, transmucosal, transplacental, transtracheal, ureteral, uretheral, and vaginal.

[0079] In an embodiment, the fusion protein of the invention is administered systemically in the methods described herein. In an embodiment, the fusion protein of the invention is administered locally in the methods described herein. In an embodiment, the fusion protein of the invention is administered directly to the tumor in the methods described herein, for example by injection or cannulation.

[0080] In an embodiment, the antibody or antibody fragment of the invention is administered systemically in the methods described herein. In an embodiment, the antibody or antibody fragment of the invention is administered locally in the methods described herein. In an embodiment, the antibody or antibody fragment of the invention is administered directly to the tumor in the methods described herein, for example by injection or cannulation.

[0081] In an embodiment, "determining" as used herein means experimentally determining.

[0082] All combinations of the various elements described herein are within the scope of the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0083] This invention will be better understood from the Experimental Details, which follow. However, one skilled in the art will readily appreciate that the specific methods and results discussed are merely illustrative of the invention as described more fully in the claims that follow thereafter.

EXPERIMENTAL DETAILS

[0084] The existing technologies for T-cell based immunotherapies regarding B7/CD28 family members work by blockade of the B7/CD28 family members. In contrast, the expression patterns and functions of BTNL9 and ERMAP, two members of the butyrophilin family, are different from CTLA-4 and PD-1 and their ligands, therefore, the BTNL9 pathway and the ERMAP pathway regulate the immune system at the different times and locations.

Example 1

[0085] mRNA expression of BTNL9, ERMAP and other members of the butyrophilin family in tissues and antigen-presenting cells: The butyrophilin family is related to the B7 family, but their expression and functions in the immune system are largely unknown. Using Real-Time RT-PCR, it was determined that BTNL9 and ERMAP, together with other family members, were widely expressed in many tissues and antigen-presenting cells (FIG. 1).

Example 2

[0086] BTNL9, ERMAP and some other members of the butyrophilin family inhibit T cell function: It was examined whether BTNL9, ERMAP and other members of the butyrophilin family were able to regulate T-cell function using a system modified from previous studies (PNAS, 110: 9879-9884, 2013). In this system, purified T cells were activated with plate-bound mAb to CD3 and the activation of T cells was determined on days 3 and T cell-derived cytokines were determined on day 2. It was determined that BTNL9 inhibited both CD4 and CD8 T cell proliferation and cytokine production, while ERMAP inhibited CD4, but not CD8 T cell proliferation (FIG. 2).

Example 3

[0087] Activated T cells have receptors for BTNL9, ERMAP and some other members of the butyrophilin family: Because BTNL9 and ERMAP inhibited T cell function, it was examined if activated T cells express receptors for these molecules. It was determined that BTNL9-Ig, BTNL9, ERMAP-Ig and other Ig fusion proteins of some members of the butyrophilin family bound to activated CD4 and CD8 T cells which were activated for three days (FIG. 3).

Example 4

[0088] The IgV1 domain of Human BTNL2 is the functional domain. As shown in FIG. 4, the IgV1 domain of human BTNL2 bound human CD8 T cells, human B cells, and human NK cells. BTNL2-IgV1-Ig protein (open histograms) and control Ig (shaded histograms) are shown in FACS assays. Sequence shown is SEQ ID NO:5.

Example 5

[0089] The IgV1 domain of Human BTNL9 is the functional domain. As shown in FIG. 5, the IgV1 domain of human BTNL9 bound human CD8 T cells, human B cells, and human NK cells. BTNL9-IgV1-Ig protein (open histograms) and control Ig (shaded histograms) are shown in FACS assays. Sequence shown is SEQ ID NO:6.

Sequence CWU 1

1

81535PRTHomo sapiens 1Met Val Asp Leu Ser Val Ser Pro Asp Ser Leu Lys Pro Val Ser Leu 1 5 10 15 Thr Ser Ser Leu Val Phe Leu Met His Leu Leu Leu Leu Gln Pro Gly 20 25 30 Glu Pro Ser Ser Glu Val Lys Val Leu Gly Pro Glu Tyr Pro Ile Leu 35 40 45 Ala Leu Val Gly Glu Glu Val Glu Phe Pro Cys His Leu Trp Pro Gln 50 55 60 Leu Asp Ala Gln Gln Met Glu Ile Arg Trp Phe Arg Ser Gln Thr Phe 65 70 75 80 Asn Val Val His Leu Tyr Gln Glu Gln Gln Glu Leu Pro Gly Arg Gln 85 90 95 Met Pro Ala Phe Arg Asn Arg Thr Lys Leu Val Lys Asp Asp Ile Ala 100 105 110 Tyr Gly Ser Val Val Leu Gln Leu His Ser Ile Ile Pro Ser Asp Lys 115 120 125 Gly Thr Tyr Gly Cys Arg Phe His Ser Asp Asn Phe Ser Gly Glu Ala 130 135 140 Leu Trp Glu Leu Glu Val Ala Gly Leu Gly Ser Asp Pro His Leu Ser 145 150 155 160 Leu Glu Gly Phe Lys Glu Gly Gly Ile Gln Leu Arg Leu Arg Ser Ser 165 170 175 Gly Trp Tyr Pro Lys Pro Lys Val Gln Trp Arg Asp His Gln Gly Gln 180 185 190 Cys Leu Pro Pro Glu Phe Glu Ala Ile Val Trp Asp Ala Gln Asp Leu 195 200 205 Phe Ser Leu Glu Thr Ser Val Val Val Arg Ala Gly Ala Leu Ser Asn 210 215 220 Val Ser Val Ser Ile Gln Asn Leu Leu Leu Ser Gln Lys Lys Glu Leu 225 230 235 240 Val Val Gln Ile Ala Asp Val Phe Val Pro Gly Ala Ser Ala Trp Lys 245 250 255 Ser Ala Phe Val Ala Thr Leu Pro Leu Leu Leu Val Leu Ala Ala Leu 260 265 270 Ala Leu Gly Val Leu Arg Lys Gln Arg Arg Ser Arg Glu Lys Leu Arg 275 280 285 Lys Gln Ala Glu Lys Arg Gln Glu Lys Leu Thr Ala Glu Leu Glu Lys 290 295 300 Leu Gln Thr Glu Leu Asp Trp Arg Arg Ala Glu Gly Gln Ala Glu Trp 305 310 315 320 Arg Ala Ala Gln Lys Tyr Ala Val Asp Val Thr Leu Asp Pro Ala Ser 325 330 335 Ala His Pro Ser Leu Glu Val Ser Glu Asp Gly Lys Ser Val Ser Ser 340 345 350 Arg Gly Ala Pro Pro Gly Pro Ala Pro Gly His Pro Gln Arg Phe Ser 355 360 365 Glu Gln Thr Cys Ala Leu Ser Leu Glu Arg Phe Ser Ala Gly Arg His 370 375 380 Tyr Trp Glu Val His Val Gly Arg Arg Ser Arg Trp Phe Leu Gly Ala 385 390 395 400 Cys Leu Ala Ala Val Pro Arg Ala Gly Pro Ala Arg Leu Ser Pro Ala 405 410 415 Ala Gly Tyr Trp Val Leu Gly Leu Trp Asn Gly Cys Glu Tyr Phe Val 420 425 430 Leu Ala Pro His Arg Val Ala Leu Thr Leu Arg Val Pro Pro Arg Arg 435 440 445 Leu Gly Val Phe Leu Asp Tyr Glu Ala Gly Glu Leu Ser Phe Phe Asn 450 455 460 Val Ser Asp Gly Ser His Ile Phe Thr Phe His Asp Thr Phe Ser Gly 465 470 475 480 Ala Leu Cys Ala Tyr Phe Arg Pro Arg Ala His Asp Gly Gly Glu His 485 490 495 Pro Asp Pro Leu Thr Ile Cys Pro Leu Pro Val Arg Gly Thr Gly Val 500 505 510 Pro Glu Glu Asn Asp Ser Asp Thr Trp Leu Gln Pro Tyr Glu Pro Ala 515 520 525 Asp Pro Ala Leu Asp Trp Trp 530 535 2475PRTHomo sapiens 2Met Glu Met Ala Ser Ser Ala Gly Ser Trp Leu Ser Gly Cys Leu Ile 1 5 10 15 Pro Leu Val Phe Leu Arg Leu Ser Val His Val Ser Gly His Ala Gly 20 25 30 Asp Ala Gly Lys Phe His Val Ala Leu Leu Gly Gly Thr Ala Glu Leu 35 40 45 Leu Cys Pro Leu Ser Leu Trp Pro Gly Thr Val Pro Lys Glu Val Arg 50 55 60 Trp Leu Arg Ser Pro Phe Pro Gln Arg Ser Gln Ala Val His Ile Phe 65 70 75 80 Arg Asp Gly Lys Asp Gln Asp Glu Asp Leu Met Pro Glu Tyr Lys Gly 85 90 95 Arg Thr Val Leu Val Arg Asp Ala Gln Glu Gly Ser Val Thr Leu Gln 100 105 110 Ile Leu Asp Val Arg Leu Glu Asp Gln Gly Ser Tyr Arg Cys Leu Ile 115 120 125 Gln Val Gly Asn Leu Ser Lys Glu Asp Thr Val Ile Leu Gln Val Ala 130 135 140 Ala Pro Ser Val Gly Ser Leu Ser Pro Ser Ala Val Ala Leu Ala Val 145 150 155 160 Ile Leu Pro Val Leu Val Leu Leu Ile Met Val Cys Leu Cys Leu Ile 165 170 175 Trp Lys Gln Arg Arg Ala Lys Glu Lys Leu Leu Tyr Glu His Val Thr 180 185 190 Glu Val Asp Asn Leu Leu Ser Asp His Ala Lys Glu Lys Gly Lys Leu 195 200 205 His Lys Ala Val Lys Lys Leu Arg Ser Glu Leu Lys Leu Lys Arg Ala 210 215 220 Ala Ala Asn Ser Gly Trp Arg Arg Ala Arg Leu His Phe Val Ala Val 225 230 235 240 Thr Leu Asp Pro Asp Thr Ala His Pro Lys Leu Ile Leu Ser Glu Asp 245 250 255 Gln Arg Cys Val Arg Leu Gly Asp Arg Arg Gln Pro Val Pro Asp Asn 260 265 270 Pro Gln Arg Phe Asp Phe Val Val Ser Ile Leu Gly Ser Glu Tyr Phe 275 280 285 Thr Thr Gly Cys His Tyr Trp Glu Val Tyr Val Gly Asp Lys Thr Lys 290 295 300 Trp Ile Leu Gly Val Cys Ser Glu Ser Val Ser Arg Lys Gly Lys Val 305 310 315 320 Thr Ala Ser Pro Ala Asn Gly His Trp Leu Leu Arg Gln Ser Arg Gly 325 330 335 Asn Glu Tyr Glu Ala Leu Thr Ser Pro Gln Thr Ser Phe Arg Leu Lys 340 345 350 Glu Pro Pro Arg Cys Val Gly Ile Phe Leu Asp Tyr Glu Ala Gly Val 355 360 365 Ile Ser Phe Tyr Asn Val Thr Asn Lys Ser His Ile Phe Thr Phe Thr 370 375 380 His Asn Phe Ser Gly Pro Leu Arg Pro Phe Phe Glu Pro Cys Leu His 385 390 395 400 Asp Gly Gly Lys Asn Thr Ala Pro Leu Val Ile Cys Ser Glu Leu His 405 410 415 Lys Ser Glu Glu Ser Ile Val Pro Arg Pro Glu Gly Lys Gly His Ala 420 425 430 Asn Gly Asp Val Ser Leu Lys Val Asn Ser Ser Leu Leu Pro Pro Lys 435 440 445 Ala Pro Glu Leu Lys Asp Ile Ile Leu Ser Leu Pro Pro Asp Leu Gly 450 455 460 Pro Ala Leu Gln Glu Leu Lys Ala Pro Ser Phe 465 470 475 3456PRTHomo sapiens 3Met Val Asp Cys Pro Arg Tyr Ser Leu Ser Gly Val Ala Ala Ser Phe 1 5 10 15 Leu Phe Val Leu Leu Thr Ile Lys His Pro Asp Asp Phe Arg Val Val 20 25 30 Gly Pro Asn Leu Pro Ile Leu Ala Lys Val Gly Glu Asp Ala Leu Leu 35 40 45 Thr Cys Gln Leu Leu Pro Lys Arg Thr Thr Ala His Met Glu Val Arg 50 55 60 Trp Tyr Arg Ser Asp Pro Ala Met Pro Val Ile Met Tyr Arg Asp Gly 65 70 75 80 Ala Val Val Thr Gly Leu Pro Met Glu Gly Tyr Gly Gly Arg Ala Glu 85 90 95 Trp Met Glu Asp Ser Thr Glu Glu Gly Ser Val Ala Leu Lys Ile Arg 100 105 110 Gln Val Gln Pro Ser Asp Asp Gly Gln Tyr Trp Cys Arg Phe Gln Glu 115 120 125 Gly Asp Tyr Trp Arg Glu Thr Ser Val Leu Leu Gln Val Ala Ala Leu 130 135 140 Gly Ser Ser Pro Asn Ile His Val Glu Gly Leu Gly Glu Gly Glu Val 145 150 155 160 Gln Leu Val Cys Thr Ser Arg Gly Trp Phe Pro Glu Pro Glu Val His 165 170 175 Trp Glu Gly Ile Trp Gly Glu Lys Leu Met Ser Phe Ser Glu Asn His 180 185 190 Val Pro Gly Glu Asp Gly Leu Phe Tyr Val Glu Asp Thr Leu Met Val 195 200 205 Arg Asn Asp Ser Val Glu Thr Ile Ser Cys Phe Ile Tyr Ser His Gly 210 215 220 Leu Arg Glu Thr Gln Glu Ala Thr Ile Ala Leu Ser Glu Arg Leu Gln 225 230 235 240 Thr Glu Leu Val Ser Val Ser Val Ile Gly His Ser Gln Pro Ser Pro 245 250 255 Val Gln Val Gly Glu Asn Ile Glu Leu Thr Cys His Leu Ser Pro Gln 260 265 270 Thr Asp Ala Gln Asn Leu Glu Val Arg Trp Leu Arg Ser Arg Tyr Tyr 275 280 285 Pro Ala Val His Val Tyr Ala Asn Gly Thr His Val Ala Gly Glu Gln 290 295 300 Met Val Glu Tyr Lys Gly Arg Thr Ser Leu Val Thr Asp Ala Ile His 305 310 315 320 Glu Gly Lys Leu Thr Leu Gln Ile His Asn Ala Arg Thr Ser Asp Glu 325 330 335 Gly Gln Tyr Arg Cys Leu Phe Gly Lys Asp Gly Val Tyr Gln Glu Ala 340 345 350 Arg Val Asp Val Gln Val Thr Ala Val Gly Ser Thr Pro Arg Ile Thr 355 360 365 Arg Glu Val Leu Lys Asp Gly Gly Met Gln Leu Arg Cys Thr Ser Asp 370 375 380 Gly Trp Phe Pro Arg Pro His Val Gln Trp Arg Asp Arg Asp Gly Lys 385 390 395 400 Thr Met Pro Ser Phe Ser Glu Ala Phe Gln Gln Gly Ser Gln Glu Leu 405 410 415 Phe Gln Val Glu Thr Leu Leu Leu Val Thr Asn Gly Ser Met Val Asn 420 425 430 Val Thr Cys Ser Ile Ser Leu Pro Leu Gly Gln Glu Lys Thr Ala Arg 435 440 445 Phe Pro Leu Ser Asp Ser Lys Ile 450 455 4260PRTHomo sapiens 4Met Val Asp Cys Pro Arg Tyr Ser Leu Ser Gly Val Ala Ala Ser Phe 1 5 10 15 Leu Phe Val Leu Leu Thr Ile Lys His Pro Asp Asp Phe Arg Val Val 20 25 30 Gly Pro Asn Leu Pro Ile Leu Ala Lys Val Gly Glu Asp Ala Leu Leu 35 40 45 Thr Cys Gln Leu Leu Pro Lys Arg Thr Thr Ala His Met Glu Val Arg 50 55 60 Trp Tyr Arg Ser Asp Pro Ala Met Pro Val Ile Met Tyr Arg Asp Gly 65 70 75 80 Ala Val Val Thr Gly Leu Pro Met Glu Gly Tyr Gly Gly Arg Ala Glu 85 90 95 Trp Met Glu Asp Ser Thr Glu Glu Gly Ser Val Ala Leu Lys Ile Arg 100 105 110 Gln Val Gln Pro Ser Asp Asp Gly Gln Tyr Trp Cys Arg Phe Gln Glu 115 120 125 Gly Asp Tyr Trp Arg Glu Thr Ser Val Leu Leu Gln Val Ala Ala Leu 130 135 140 Gly Ser Ser Pro Asn Ile His Val Glu Gly Leu Gly Glu Gly Glu Val 145 150 155 160 Gln Leu Val Cys Thr Ser Arg Gly Trp Phe Pro Glu Pro Glu Val His 165 170 175 Trp Glu Gly Ile Trp Gly Glu Lys Leu Met Ser Phe Ser Glu Asn His 180 185 190 Val Pro Gly Glu Asp Gly Leu Phe Tyr Val Glu Asp Thr Leu Met Val 195 200 205 Arg Asn Asp Ser Val Glu Thr Ile Ser Cys Phe Ile Tyr Ser His Gly 210 215 220 Leu Arg Glu Thr Gln Glu Ala Thr Ile Ala Leu Ser Glu Arg Leu Gln 225 230 235 240 Thr Glu Leu Val Ser Val Ser Val Ile Gly His Ser Gln Pro Ser Pro 245 250 255 Val Gln Val Gly 260 5129PRTHomo sapiens 5Lys Gln Ser Glu Asp Phe Arg Val Ile Gly Pro Ala His Pro Ile Leu 1 5 10 15 Ala Gly Val Gly Glu Asp Ala Leu Leu Thr Cys Gln Leu Leu Pro Lys 20 25 30 Arg Thr Thr Met His Val Glu Val Arg Trp Tyr Arg Ser Glu Pro Ser 35 40 45 Thr Pro Val Phe Val His Arg Asp Gly Val Glu Val Thr Glu Met Gln 50 55 60 Met Glu Glu Tyr Arg Gly Trp Val Glu Trp Ile Glu Asn Gly Ile Ala 65 70 75 80 Lys Gly Asn Val Ala Leu Lys Ile His Asn Ile Gln Pro Ser Asp Asn 85 90 95 Gly Gln Tyr Trp Cys His Phe Gln Asp Gly Asn Tyr Cys Gly Glu Thr 100 105 110 Ser Leu Leu Leu Lys Val Ala Gly Leu Gly Ser Ala Pro Ser Ile His 115 120 125 Met 6124PRTHomo sapiens 6Glu Val Lys Val Leu Gly Pro Glu Tyr Pro Ile Leu Ala Leu Val Gly 1 5 10 15 Glu Glu Val Glu Phe Pro Cys His Leu Trp Pro Gln Leu Asp Ala Gln 20 25 30 Gln Met Glu Ile Arg Trp Phe Arg Ser Gln Thr Phe Asn Val Val His 35 40 45 Leu Tyr Gln Glu Gln Gln Glu Leu Pro Gly Arg Gln Met Pro Ala Phe 50 55 60 Arg Asn Arg Thr Lys Leu Val Lys Asp Asp Ile Ala Tyr Gly Ser Val 65 70 75 80 Val Leu Gln Leu His Ser Ile Ile Pro Ser Asp Lys Gly Thr Tyr Gly 85 90 95 Cys Arg Phe His Ser Asp Asn Phe Ser Gly Glu Ala Leu Trp Glu Leu 100 105 110 Glu Val Ala Gly Leu Gly Ser Asp Pro His Leu Ser 115 120 75PRTArtificial Sequenceartificial linker sequenceMISC_FEATURE(1)..(5)sequence can be single or repeat 2, 3, 4, or 5 times 7Gly Gly Gly Gly Ser 1 5 85PRTArtificial Sequenceartificial linker sequenceMISC_FEATURE(1)..(5)sequence can be single or repeat 2, 3, 4, or 5 times 8Glu Ala Ala Ala Lys 1 5

Claims

1. A method of treating a tumor in a subject comprising administering to the subject an amount of a BTNL9-binding antibody, or BTNL9-binding fragment thereof, sufficient to inhibit a BTNL9 and treat the tumor.

2. A method of treating a tumor in a subject comprising administering to the subject an amount of a ERMAP-binding antibody, or ERMAP-binding fragment thereof, sufficient to inhibit a ERMAP and treat the tumor.

3. A method of treating a tumor in a subject comprising administering to the subject an amount of a BTNL2-binding antibody, or BTNL2-binding fragment thereof, sufficient to inhibit a BTNL2 and treat the tumor.

4. The method of claim 1, wherein the BTNL9 is a human BTNL9.

5. The method of claim 2, wherein the ERMAP is a human ERMAP.

6. The method of claim 3, wherein the BTNL2 is a human BTNL2.

7. The method of claim 1, wherein the tumor is a tumor of a breast, lung, thyroid, melanoma, pancreas, ovary, liver, bladder, colon, prostate, kidney, esophagus, or is a hematological tumor, or wherein the tumor is a lymphoid organ tumor.

8. The method of claim 1, wherein the antibody is administered as an adjunct to an additional anti-cancer therapy for the tumor.

9. The method of claim 1, wherein the amount of a BTNL9-binding antibody is administered.

10. The method of claim 3, wherein the amount of a BTNL2-binding antibody is administered.

11. The method of claim 1, wherein the fragment of the antibody is administered.

12-15. (canceled)

16. The method of claim 15, wherein the BTNL-2 comprises SEQ ID NO:4 but does not comprise SEQ ID NO:3.

17. The method of claim 13, wherein the plasma-soluble BTNL9, or the plasma-soluble ERMAP, respectively, comprises an BTNL9 fused to an immunoglobulin polypeptide, or an ERMAP fused to an immunoglobulin polypeptide, respectively.

18. The method of claim 15, wherein the plasma-soluble BTNL2 comprises an BTNL2 fused to an immunoglobulin polypeptide.

19. The method of claim 17, wherein the immunoglobulin polypeptide comprises an Fc portion of an immunoglobulin G.

20-26. (canceled)