Anti-BTLA antibodies

Abstract

The present invention generally relates to antibodies or antigen-binding fragments that bind to human B and T lymphocyte attenuator (BTLA) and their uses. More specifically, the present invention relates to agonistic antibodies that bind to human BTLA and modulate its activity, and their use in treating inflammatory, autoimmune and proliferative diseases and disorders.

Description

[0001] Cross-references

[0002] This application claims the benefit of UK application No. 1820554.2, filed on December 17, 2018, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention generally relates to antibodies or antigen-binding fragments that bind to human B and T lymphocyte attenuator (BTLA) and their uses. More specifically, the present invention relates to agonistic antibodies that bind to human BTLA and modulate its activity, and their use in treating inflammatory, autoimmune and proliferative diseases and disorders. Background Art

[0004] The immune system must strike a balance between the destruction of pathogens or dangerous mutant cells and the tolerance of healthy self-tissues and harmless commensals. To promote this balance, the activity of immune cells is influenced by the integration of signals from a variety of stimulatory and inhibitory receptors that adapt the cells to their environment. These surface-expressed receptors present attractive targets for therapeutic modulation of immune responses. Many human diseases are caused by abnormal or unwanted activation of the immune system, including autoimmune diseases, transplant rejection, and graft-versus-host disease. Agonists that can induce signaling through inhibitory receptors can suppress these unwanted immune responses.

[0005] B and T lymphocyte attenuator (BTLA; also known as CD272) is an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS and PD-1 (Watanabe et al., Nat Immunol. 4:670-679, 2003). It is widely expressed on myeloid and lymphoid cells throughout the immune system (Han et al., J Immunol. 172:5931-9, 2004). After engagement of its ligand, herpes virus entry mediator (HVEM), BTLA recruits the phosphatases SHP-1 and SHP-2 to its cytoplasmic domain (Sedy et al., Nat Immunol. 6:90-8, 2005), thereby inhibiting the signaling cascade that activates the receptor. Mice lacking the complete BTLA gene exhibited hyperproliferative B and T cell responses in vitro, with higher titers to DNP-KLH after immunization and increased sensitivity to EAE (Watanabe et al., Nat. Immunol, 4:670-679, 2003). BTLA knockout mice spontaneously produce autoantibodies, autoimmune hepatitis-like diseases, and inflammatory cell infiltration of multiple organs, if not observed until old age (Oya et al., Arthritis Rheum 58:2498-2510, 2008). This evidence suggests that BTLA inhibitory receptors play a crucial role in maintaining immune homeostasis and suppressing autoimmunity. In addition, HVEM-BTLA signaling is involved in the regulation of mucosal inflammation and infection immunity (Shui et al., J Leukoc Biol. 89:517-523, 2011).

[0006] There is a great need for therapeutic agents that can modulate BTLA function to suppress autoreactive lymphocytes in the setting of autoimmune diseases.

[0007] It has been previously shown that monoclonal antibodies that bind to mouse BTLA can act as agonists, inducing signaling through the receptor to inhibit immune cell responses. In the presence of agonist anti-BTLA antibodies (mAbs), anti-CD3 and anti-CD28 activated T cells showed reduced IL-2 production and proliferation (Kreig et al., J. Immunol., 175, 6420-6472, 2005).

[0008] In addition, anti-mouse BTLA agonist antibodies have been shown to improve disease in a mouse model of graft-versus-host disease (Sakoda et al., Blood. 117:2506-2514; Albring et al., J Exp Med. 207:2551-9, 2010). Agonist antibodies targeting the human BTLA receptor have been shown to inhibit T cell responses in vitro (see Otsuki et al., Biochem Biophys Res Commun 344:1121-7, 2006; and WO 2011 / 014438), but have not yet been translated into clinical application.

[0009] There is a need in the art to discover new and useful agents, such as antibodies or antigen-binding antibody fragments, that can modulate BTLA. Summary of the Invention

[0010] The present invention relates to anti-human BTLA agonist antibodies or antibody fragments thereof having one or more desirable properties, including high binding affinity, high agonist potency, high agonist efficacy, good pharmacokinetics, and low antigenicity in human subjects. The present invention also relates to the use of the antibodies or antibody fragments of the present invention in treating diseases.

[0011] According to a first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds BTLA is provided, wherein the antibody has a heavy chain and / or light chain having at least one complementarity determining region (CDR) present in an antibody selected from the group consisting of 11.5.1, 2.8.6, 12F11, 14D4, 15B6, 15C6, 16E1, 16F10, 16H2, 1H6, 21C7, 24H7, 26B1, 26F3, 27G9, 3A9, 3E8, 4B1, 4D3, 4D5, 4E8, 4H4, 6G8, 7A1, 8B4, 8C4, 6.2 and 831, as identified in Table 1 and described herein.

[0012] According to a second aspect of the present invention, an isolated nucleic acid is provided, comprising a nucleotide sequence encoding a heavy chain polypeptide or a light chain polypeptide of the isolated antibody or antigen-binding fragment thereof according to the first aspect of the present invention.

[0013] According to a third aspect of the present invention, there is provided a vector comprising the nucleic acid of the second aspect of the present invention.

[0014] According to a fourth aspect of the present invention, a host cell comprising the nucleic acid sequence according to the second aspect of the present invention or the vector according to the third aspect of the present invention is provided.

[0015] According to a fifth aspect of the present invention, a method for producing an antibody or antigen-binding fragment thereof according to the first aspect of the present invention is provided, comprising the steps of culturing the host cell of the fourth aspect of the present invention under conditions for producing the antibody or antigen-binding fragment thereof, and optionally isolating and / or purifying the antibody or antigen-binding fragment thereof.

[0016] According to the sixth aspect of the present invention, a pharmaceutical composition is provided, comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the antibody or antigen-binding fragment thereof of the first aspect of the present invention, or the antibody or antigen-binding fragment thereof produced by the fifth aspect of the present invention.

[0017] According to a seventh aspect of the present invention, there is provided a method for preparing a pharmaceutical composition, the method comprising formulating the antibody or antigen-binding fragment thereof according to the first aspect of the present invention, or the antibody or antigen-binding fragment thereof produced according to the fifth aspect, into a composition comprising at least one additional component. In a particular embodiment, the at least one additional component is a pharmaceutically acceptable excipient.

[0018] According to an eighth aspect of the present invention, there is provided a method for treating a BTLA-related disease in a patient, comprising administering to the patient a therapeutically effective amount of the antibody or antigen-binding fragment thereof according to the first aspect of the present invention or the pharmaceutical composition according to the sixth aspect of the present invention. DETAILED DESCRIPTION

[0019] The present inventors have identified particularly potent agonist antibodies to BTLA that are expected to be more effective than current antibodies in suppressing T cell responses and therefore particularly useful in treating immune-mediated disorders.

[0020] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a molecule" optionally includes a combination of two or more such molecules, and so forth.

[0021] It should be understood that wherever aspects are described herein with the language "comprising," other similar aspects described with the language "consisting of" and / or "consisting essentially of" are also provided.

[0022] It should be understood that, unless the context clearly provides otherwise, one, some or all of the properties of the various embodiments described herein may be applied to any aspect. In addition, the various embodiments may be combined to form other embodiments of the present invention. These and other aspects of the present invention will become apparent to those skilled in the art. These and other embodiments of the present invention are further described by the following detailed description.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure relates. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd edition, 1999, Academic Press; and Oxford Dictionary of Biochemistry and Molecular Biology, Revised Edition, 2000, Oxford University Press, provide a general dictionary of many of the terms used in the present disclosure for one of ordinary skill in the art.

[0024] As used herein, the term "about" refers to the typical error range of the corresponding value that is readily known to those skilled in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments involving the value or parameter itself.

[0025] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Likewise, nucleotides may be referred to by their commonly accepted single-letter codes.

[0026] Unless otherwise indicated, amino acid numbering in the variable domains, CDRs, and framework regions (FRs) of antibodies follows the Kabat definition as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991).

[0027] The terms "B and T lymphocyte attenuator" and "BTLA" are used interchangeably and refer to a protein or gene (or other nucleic acid encoding all or part of BTLA), unless the context indicates otherwise. The human BTLA sequence includes all human isoforms and variant forms. A representative example of full-length human BTLA is disclosed in Genbank under accession number AJ717664.1. Another representative polypeptide sequence of human BTLA is disclosed in SEQ ID NO: 23, which differs from the sequence in AJ717664.1 only in two naturally occurring variant single nucleotide polymorphisms. Despite allelic variation, the human BTLA polypeptide sequence typically has at least 90% sequence identity (e.g., at least 95%, 96%, 97%, 98%, 99%, or 100%) to the human BTLA in SEQ ID NO: 23.

[0028] A representative example of full-length cynomolgus monkey (cyno) BTLA is disclosed in Genbank under the accession number XP_005548224. A reference polypeptide sequence of cynomolgus monkey BTLA is disclosed in SEQ ID NO: 24. The cynomolgus monkey BTLA polypeptide sequence typically has at least 90% sequence identity (e.g., at least 95%, 96%, 97%, 98%, 99%, or 100%) to the cynomolgus monkey BTLA disclosed in SEQ ID NO: 24.

[0029] The term sequence identity is well known in the art. For the purposes of the present invention, when determining whether a target sequence meets a defined limit (e.g., 90% identity), it is considered to meet the defined limit if it is identified as meeting the defined limit using the BLAST (Basic Local Alignment Search Tool) algorithm (see Altschul et al. J Mol Biol 215:403-410, 1990) or the Smith-Waterman algorithm (see Smith and Waterman. J Mol. Biol. 147:195-197, 1981).

[0030] Antibodies and Antigen-Binding Fragments of Antibodies

[0031] Antibodies are immunoglobulin molecules that are capable of specifically binding to targets such as proteins, polypeptides, peptides, carbohydrates, polynucleotides, lipids, or combinations thereof, through at least one antigen recognition site located in the variable domain of the immunoglobulin molecule. In particular, as used herein, the term "antibody" includes intact polyclonal antibodies, intact monoclonal antibodies, multispecific antibodies such as bispecific antibodies generated from at least two intact antibodies, chimeric antibodies, humanized antibodies, human antibodies, and any other modified immunoglobulin molecules containing an antigen recognition site, as long as the antibody exhibits the desired biological activity.

[0032] The term "antibody" as used herein refers to an immunoglobulin molecule that specifically binds an antigen and includes an FcR binding site that may or may not be functional.

[0033] As used herein, a BTLA agonist antibody (or antibody fragment) refers to an antibody (or antibody fragment) that can bind to BTLA and enhance its co-inhibitory signal on T and / or B cells.

[0034] Antigen binding site refers to the part of a molecule that binds to all or part of the target antigen. In an antibody molecule, it can be referred to as the antigen binding site of the antibody, and includes the part of the antibody that specifically binds to all or part of the target antigen. When the antigen is large, the antibody may only bind to a specific part of the antigen, which is called an epitope. The antigen binding site of an antibody can be provided by one or more antibody variable domains. Preferably, the antigen binding site of an antibody comprises an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH).

[0035] The present invention also includes antibody fragments that contain an antigen-binding site. Thus, when referring to an antibody, the term "antigen-binding fragment thereof" refers to antibody fragments, such as Fab, Fab', F(ab')2, diabodies, Fv fragments, and single-chain Fv (scFv) mutants, that have an antigen recognition site and, therefore, have the ability to bind to an antigen. Antigen-binding immunoglobulin (antibody) fragments are well known in the art. Such fragments do not necessarily have a functional Fc receptor binding site.

[0036] As used herein, the terms "antibody fragment molecule of the invention," "antibody fragment," and "antigen-binding fragment thereof" are used interchangeably herein.

[0037] As used herein, the term "BTLA-binding molecule" refers to antibodies and binding fragments thereof that are capable of binding to BTLA.

[0038] There are five major immunoglobulin classes (i.e., isotypes): IgA, IgD, IgE, IgG, and IgM, and several of them can be further divided into subclasses (subtypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the different classes of immunoglobulins are referred to as α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known. Unless otherwise specified by the contextual limitations, the antibodies of the present invention may be from one of these classes or subclasses of antibodies. The heavy chain constant domains corresponding to the different antibody classes are generally represented by the corresponding lowercase Greek letters α, δ, ε, γ, and μ, respectively. The light chains of antibodies from any vertebrate species can be classified into one of two distinct types, referred to as κ (kappa) and λ (lambda), based on the amino acid sequence of their constant domains.

[0039] "Native antibodies" are typically heterotetrameric, Y-shaped glycoproteins of approximately 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, with the number of disulfide bond variations among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bond. Each heavy chain has a variable domain (VH) at one end followed by a number of constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at the other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Specific amino acid residues are believed to form the interface between the light and heavy chain variable domains. Each heavy chain contains a variable domain (VH) and a constant region, which in the case of IgG, IgA, and IgD antibodies consists of three domains, referred to as C, D, and E. H 1. C H 2 and C H 3 (IgM and IgE have a fourth domain - C H 4). In the IgG, IgA and IgD classes, C H 1 and C H The two domains are separated by a flexible hinge region, which is a proline and cysteine-rich segment of variable length (about 10 to about 60 amino acids in various IgG subclasses). The variable domains in both the light and heavy chains are connected to the constant domains by a "J" region of about 12 or more amino acids, and the heavy chain also has a "D" region of about 10 additional amino acids. Each class of antibodies further comprises interchain and intrachain disulfide bonds formed by paired cysteine ​​residues. The heavy chain variable region (VH) and the light chain variable region (VL) can each be further subdivided into hypervariable regions, referred to as CDRs, interspersed with more conserved regions, referred to as framework regions (FRs). Each VH and VL comprises three CDRs and four FRs, arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains comprise binding domains that interact with antigens. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.

[0040] The antibodies or antigen-binding fragments thereof of the present invention can be from any animal species, including mouse, rat, human or any other source (including chimeric or humanized antibodies). In some embodiments, the antibody or antigen-binding fragment is monoclonal, such as a monoclonal antibody. In some embodiments, the antibody or antigen-binding fragment is a human or humanized antibody or antigen-binding fragment thereof. Non-human antibodies or antigen-binding fragments thereof can be humanized by recombinant methods that reduce their immunogenicity in humans.

[0041] The antibodies or antigen-binding fragments thereof of the present invention can be identified using known methods. For example, the antigen-binding portion may have been selected using phage display or other antigen-binding selection or panning methods. Such antigen-binding portions can then be incorporated into an antibody framework (e.g., fused to the constant region and hinge region of, for example, an IgG1 or IgG4 molecule).

[0042] As used herein, the term "monoclonal antibody" ("mAb") refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible mutations, such as naturally occurring mutations, that may be present in minor amounts. Thus, the modifier "monoclonal" indicates the character of the antibody or fragment thereof as not being a mixture of discrete antibodies or antigen-binding fragments. mAbs are highly specific, being directed against a single antigenic site / epitope.

[0043] mAbs can be produced by hybridoma, recombination, transgenic or other techniques known to those skilled in the art. For example, monoclonal antibodies or antigen-binding fragments thereof according to the present invention can be prepared by the hybridoma method first described by Kohler and Milstein (Nature 256:495, 1975), or can be prepared by recombinant DNA methods as described in U.S. Patents 4,816,567 and 6,331,415. For example, "monoclonal antibodies" can also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 1991; 352:624-628 and Marks et al., J. Mol. Biol. 1991; 222:581-597.

[0044] The term monoclonal may also be applied to the antigen-binding fragments of the antibodies of the invention. It simply means that the molecule is produced or exists in a single clone.

[0045] "HuMAb" refers to an antibody having a variable region in which the framework and CDR regions are derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. Human antibodies may include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced in vitro by random or site-specific mutagenesis or in vivo by somatic mutation). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been transplanted onto human framework sequences.

[0046] Human antibodies can be prepared by administering immunogens / antigens to transgenic animals that have been modified to produce complete human antibodies or complete antibodies with human variable regions in response to antigenic attack, but whose endogenous loci have been disabled, such as immunized xenogeneic mice (xenomice) (see, for example, U.S. Patents 6,075,181 and 6,150,584 about XENOMOUSE (trademark) technology). See also, for example, Li et al. (Proc. Natl. Acad. Sci. USA 103:3557-3562, 2006), which relate to human antibodies produced by human B cell hybridoma technology. Such animals generally contain all or part of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or are present outside the chromosomes or randomly integrated into the chromosomes of the animal. In such transgenic mice, the endogenous immunoglobulin loci are usually inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23: 1117-1125 (2005). See also, for example, the description of the XENOMOUSE TM U.S. Patents 6,075,181 and 6,150,584; describing HUMAB TM US Patent 5,770,429; describes the K-MMOUSE TM US Patent 7,041,870, and describes the VELOCIMOUSE TM The technology is disclosed in US Patent Application Publication No. US2007 / 0061900. Human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

[0047] Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor, J. Immunol, 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol, 147: 86 (1991)). Human antibodies generated by human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Other methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26:265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20:927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27:185-91 (2005).

[0048] The terms "human" antibody and "fully human" antibody are used synonymously. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues.

[0049] As used herein, " humanized antibody " refers to the antibody that some, most or all of the amino acids outside the CDR of wherein non-human antibody are replaced by the corresponding amino acids derived from human immunoglobulin.In some embodiments, humanized antibody is human immunoglobulin (acceptor antibody), wherein the residue from the CDR of acceptor is replaced by the residue from the CDR of non-human species (donor antibody) such as mouse, rat or rabbit with required specificity, affinity and ability.Humanized antibody can be included in the CDR or framework sequence of acceptor antibody and all do not find but be included to further improve and optimize the residue of antibody performance.In one embodiment of the humanized form of antibody, some, most or all of the amino acids outside the CDR are replaced by the amino acids from human immunoglobulin, and some, most or all of the amino acids in one or more CDR regions are unchanged.Amino acid whose small addition, deletion, insertion, replacement or modification are allowed, provided that they can not eliminate the ability of antibody to be combined with specific antigen. "Humanized" antibody retains the antigenic specificity similar to the original antibody. Typically, a humanized antibody will comprise substantially all of at least one, typically two variable domains, wherein all or substantially all of the hypervariable loops correspond to the hypervariable loops of a non-human immunoglobulin, and all or substantially all of the FRs are FRs of human immunoglobulin sequences. Humanized antibodies optionally also comprise at least a portion of an immunoglobulin constant region (Fc), generally at least a portion of an immunoglobulin constant region of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1: 105-115 (1998); Harris, Biochem. Soc. Transactions 23: 1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5: 428-433 (1994); and U.S. Patents 6,982,321 and 7,087,409.

[0050] As used herein, an "engineered antibody" refers to an antibody, which may be a humanized antibody, in which specific residues have been replaced with other residues to reduce adverse effects or properties. Such replacements may be within the CD domain. For example, as described herein (see Example 21), the CDRH2 of the humanized antibody 3E8 was modified with an N57Q substitution to remove deamidation potential and a K63S substitution to reduce predicted immunogenicity.

[0051] "Chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as a variable region derived from a mouse antibody and a constant region derived from a human antibody, or vice versa. The term also includes antibodies comprising a variable region from an individual (such as a first mouse) of a species and a constant region from another individual (such as a second mouse) of the same species. The term "antigen (Ag)" refers to a molecular entity for immunizing immunocompetent vertebrates to produce antibodies (Ab) or screening expression libraries (such as phage, yeast or ribosome display libraries, etc.) identifying Ag. In this article, Ag is more broadly named, typically intended to include a target molecule specifically recognized by Ab, and is therefore included in an immune process for producing Ab or a portion or mimic of the molecule used in the library screening for selecting Ab.

[0052] "Bispecific" or "bifunctional" antibodies are artificial hybrid antibodies with two different heavy / light chain pairs and two different binding sites. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537-539 (1983)). Methods for preparing bispecific antibodies are within the capabilities of those skilled in the art. For example, bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or connection of Fab' fragments. See, for example, Songsivilai et al., (1990) Clin. Exp. Immunol. 79:315-321, Kostelny et al., (1992) J Immunol. 148:1547-1553. Additionally, bispecific antibodies can be formed as "diabodies" (Holliger et al., (1993) PNAS USA 90:6444-6448) or "Janusins" (Traunecker et al., (1991) EMBO J. 10:3655-3659 and Traunecker et al., (1992) Int. J. Cancer Suppl. 7:51-52).

[0053] Generally, the term "epitope" refers to the region or portion of an antigen to which an antibody specifically binds, i.e., the region or portion that is in physical contact with the antibody. Thus, the term "epitope" refers to a portion of a molecule that can be recognized and bound by an antibody at one or more antigen-binding regions of an antibody. Generally, an epitope is defined in the context of molecular interactions between an antibody or its antigen-binding portion (Ab) and its corresponding antigen. An epitope is typically composed of surface groupings of molecules such as amino acids or sugar side chains and has specific three-dimensional structural characteristics as well as specific charge characteristics. In some embodiments, an epitope can be a protein epitope. A protein epitope can be linear or conformational. In a linear epitope, all interaction points between a protein and an interacting molecule (such as an antibody) occur linearly along the primary amino acid sequence of the protein. A "non-linear epitope" or "conformational epitope" comprises a non-continuous polypeptide (or amino acid) within an antigenic protein to which an antibody specific for the epitope binds. As used herein, the term "antigenic epitope" is defined as a portion of an antigen to which an antibody can specifically bind, as determined by any method known in the art, such as by conventional immunoassays.

[0054] An antibody that "specifically binds" to an epitope is a term well known in the art, and methods for determining such specific binding are also well known in the art. A molecule is said to exhibit "specific binding" if it reacts or associates with a particular cell, protein, or substance more frequently, more rapidly, for a longer duration, and / or with a higher affinity than with other cells, proteins, or substances.

[0055] A variety of assay formats can be used to select antibodies or peptides that specifically bind to a molecule of interest. For example, solid phase ELISA immunoassays, immunoprecipitation, Biacore TM (GE Healthcare, Piscataway, NJ), KinExA, fluorescence-activated cell sorting (FACS), Octet TM (ForteBio, Inc., Menlo Park, CA) and Western blot analysis are among the many assays that can be used to identify antibodies that specifically react with an antigen or receptor or its ligand binding portion that specifically binds to a cognate ligand or binding partner. Typically, a specific or selective reaction will be at least twice the background signal or noise, more typically more than 10 times the background, even more typically more than 50 times the background, more typically more than 100 times the background, more typically more than 500 times the background, more typically more than 1000 times the background, and even more typically more than 10,000 times the background. In addition, when the equilibrium dissociation constant (K D An antibody is said to “specifically bind” an antigen when its KD (Kd, used interchangeably herein) is <7 nM.

[0056] According to a first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to BTLA is provided, wherein the antibody has a heavy chain and / or light chain having at least one CDR present in an antibody selected from the group consisting of 11.5.1, 2.8.6, 12F11, 14D4, 15B6, 15C6, 16E1, 16F10, 16H2, 1H6, 21C7, 24H7, 26B1, 26F3, 27G9, 3A9, 3E8, 4B1, 4D3, 4D5, 4E8, 4H4, 6G8, 7A1, 8B4, 8C4, 6.2, and 831, as disclosed in Table 1 and described herein. In one embodiment, the antibody or antigen-binding fragment thereof competes for binding to BTLA with its natural ligand, HVEM. In another embodiment, the antibody or antigen-binding fragment thereof does not interfere with the binding of HVEM.

[0057] In additional embodiments, the isolated antibody that binds human BTLA is selected from 11.5.1 and 2.8.6, wherein the antibody specifically binds BTLA and induces signaling through the receptor.

[0058] As disclosed in Table 1 and described herein, an antibody selected from 11.5.1, 2.8.6, 12F11, 14D4, 15B6, 15C6, 16E1, 16F10, 16H2, 1H6, 21C7, 24H7, 26B1, 26F3, 27G9, 3A9, 3E8, 4B1, 4D3, 4D5, 4E8, 4H4, 6G8, 7A1, 8B4, 8C4, 6.2, and 831 refers to any antibody (murine, humanized, or humanized / engineered) disclosed in Table 1. One or more, e.g., VH CDRs 1, 2, and 3, or VL CDRs 1, 2, and 3, or any antibody or antigen-binding fragment thereof.

[0059] According to variations of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising at least one VH CDR having an amino acid sequence as shown in any one of SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, and having 0 to 3 amino acid modifications, such as 0, 1, 2, or 3 amino acid modifications. In certain embodiments, the amino acid modifications include, but are not limited to, amino acid substitutions, additions, deletions, or chemical modifications, and do not abolish the antibody binding affinity or T cell inhibitory effect of the modified amino acid sequence compared to the unmodified amino acid sequence.

[0060] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VH CDR having an amino acid sequence as shown in any one of SEQ ID NO: 193, SEQ ID NO: 194 or SEQ ID NO: 195, with 0 to 3 amino acid modifications.

[0061] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO:7, CDRH2 has the amino acid sequence shown in SEQ ID NO:8, and CDRH3 has the amino acid sequence shown in SEQ ID NO:9.

[0062] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO: 193, CDRH2 has the amino acid sequence shown in SEQ ID NO: 194, and CDRH3 has the amino acid sequence shown in SEQ ID NO: 195.

[0063] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 12, with 0 to 3 amino acid modifications.

[0064] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 12, with 0 to 3 amino acid modifications.

[0065] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO: 10, CDRL2 has the amino acid sequence shown in SEQ ID NO: 11, and CDRL3 has the amino acid sequence shown in SEQ ID NO: 12.

[0066] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO: 196, CDRL2 has the amino acid sequence shown in SEQ ID NO: 197, and CDRL3 has the amino acid sequence shown in SEQ ID NO: 12.

[0067] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO:7, CDRH2 has the amino acid sequence shown in SEQ ID NO:8, and CDRH3 has the amino acid sequence shown in SEQ ID NO:9, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO:10, CDRL2 has the amino acid sequence shown in SEQ ID NO:11, and CDRL3 has the amino acid sequence shown in SEQ ID NO:12.

[0068] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence set forth in SEQ ID NO: 193, CDRH2 has the amino acid sequence set forth in SEQ ID NO: 194, and CDRH3 has the amino acid sequence set forth in SEQ ID NO: 195, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence set forth in SEQ ID NO: 196, CDRL2 has the amino acid sequence set forth in SEQ ID NO: 197, and CDRL3 has the amino acid sequence set forth in SEQ ID NO: 12.

[0069] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3, with 0 to 3 amino acid modifications.

[0070] According to another variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 199, SEQ ID NO: 200 or SEQ ID NO: 201, with 0 to 3 amino acid modifications.

[0071] According to a variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO:1, CDRH2 has the amino acid sequence shown in SEQ ID NO:2, and CDRH3 has the amino acid sequence shown in SEQ ID NO:3.

[0072] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO: 199, CDRH2 has the amino acid sequence shown in SEQ ID NO: 200, and CDRH3 has the amino acid sequence shown in SEQ ID NO: 201.

[0073] According to a variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising at least one VL CDR having an amino acid sequence as shown in SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6.

[0074] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 202, SEQ ID NO: 203 or SEQ ID NO: 6.

[0075] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO:4, CDRL2 has the amino acid sequence shown in SEQ ID NO:5, and CDRL3 has the amino acid sequence shown in SEQ ID NO:6.

[0076] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO: 202, CDRL2 has the amino acid sequence shown in SEQ ID NO: 203, and CDRL3 has the amino acid sequence shown in SEQ ID NO: 6.

[0077] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO:1, CDRH2 has the amino acid sequence shown in SEQ ID NO:2, and CDRH3 has the amino acid sequence shown in SEQ ID NO:3, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO:4, CDRL2 has the amino acid sequence shown in SEQ ID NO:5, and CDRL3 has the amino acid sequence shown in SEQ ID NO:6.

[0078] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence shown in SEQ ID NO: 199, CDRH2 has the amino acid sequence shown in SEQ ID NO: 200, and CDRH3 has the amino acid sequence shown in SEQ ID NO: 201, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO: 202, CDRL2 has the amino acid sequence shown in SEQ ID NO: 203, and CDRL3 has the amino acid sequence shown in SEQ ID NO: 6.

[0079] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising at least one VH CDR having an amino acid sequence as follows: (1) SEQ ID NO: 31, 32, or 33, with 0 to 3 amino acid modifications; (2) SEQ ID NO: 37, 38, or 39, with 0 to 3 amino acid modifications; (3) SEQ ID NO: 43, 44, or 45, with 0 to 3 amino acid modifications; (4) SEQ ID NO: 43, 56, or 57, with 0 to 3 amino acid modifications; (5) SEQ ID NO: 61, 62, or 63, with 0 to 3 amino acid modifications; (6) SEQ ID NO: 31, 32, or 69, with 0 to 3 amino acid modifications; (7) SEQ ID NO: 73, 74, or 75, with 0 to 3 amino acid modifications; (8) SEQ ID NO: 79, 80, or 81, with 0 to 3 amino acid modifications; (9) SEQ ID NO: NO: 85, 86 or 87, with 0 to 3 amino acid modifications; (10) SEQ ID NO: 61, 92 or 93, with 0 to 3 amino acid modifications; (11) SEQ ID NO: 97, 98 or 99, with 0 to 3 amino acid modifications; (12) SEQ ID NO: 103, 104 or 105, with 0 to 3 amino acid modifications; (13) SEQ ID NO: 109, 110 or 111, with 0 to 3 amino acid modifications; (14) SEQ ID NO: 85, 110 or 117, with 0 to 3 amino acid modifications; (15) SEQ ID NO: 121, 122 or 123, with 0 to 3 amino acid modifications; (16) SEQ ID NO: 127, 128 or 129, with 0 to 3 amino acid modifications; (17) SEQ ID NO: 133, 134 or 135, with 0 to 3 amino acid modifications; (18) SEQ ID NO: 139, 140 or 141, with 0 to 3 amino acid modifications; (19) SEQ ID NO: 145, 146 or 147, with 0 to 3 amino acid modifications; (20) SEQ ID NO: 31, 32 or 33, with 0 to 3 amino acid modifications; (21) SEQ ID NO: 31, 32 or 159, with 0 to 3 amino acid modifications; (22) SEQ ID NO: 169, 170 or 171, with 0 to 3 amino acid modifications; (23) SEQ ID NO: 61, 62 or 63, with 0 to 3 amino acid modifications; (24) SEQ ID NO: 31, 182 or 183, with 0 to 3 amino acid modifications; (25) SEQ ID NO: 187, 188 or 189, with 0 to 3 amino acid modifications;(26) SEQ ID NO: 193, 194 or 195, with 0 to 3 amino acid modifications; (27) SEQ ID NO: 199, 200 or 201, with 0 to 3 amino acid modifications; (28) SEQ ID NO: 205, 206 or 207, with 0 to 3 amino acid modifications; (29) SEQ ID NO: 211, 212 or 213, with 0 to 3 amino acid modifications; (30) SEQ ID NO: 127, 386 or 129, with 0 to 3 amino acid modifications; (33) SEQ ID NO: 205, 206 or 207, with 0 to 3 amino acid modifications; (34) SEQ ID NO: 127, 388 or 129, with 0 to 3 amino acid modifications; or (35) SEQ ID NO: 205, 387 or 207, with 0 to 3 amino acid modifications.

[0080] According to a variation of the first aspect of the present invention, there is provided an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1, CDRH2, and CDRH3 have the following amino acid sequences: (1) SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, respectively; (2) SEQ ID NO:37, SEQ ID NO:38, and SEQ ID NO:39, respectively; (3) SEQ ID NO:43, SEQ ID NO:44, and SEQ ID NO:45, respectively; (4) SEQ ID NO:43, SEQ ID NO:56, and SEQ ID NO:57, respectively; (5) SEQ ID NO:61, SEQ ID NO:62, and SEQ ID NO:63, respectively; (6) SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:69, respectively; (7) SEQ ID NO:73, SEQ ID NO:74, and SEQ ID NO:75, respectively. NO:75; (8) are SEQ ID NO:79, SEQ ID NO:80 and SEQ ID NO:81 respectively; (9) are SEQ ID NO:85, SEQ ID NO:86 and SEQ ID NO:87 respectively; (10) are SEQ ID NO:61, SEQ ID NO:92 and SEQ ID NO:93 respectively; (11) are SEQ ID NO:97, SEQ ID NO:98 and SEQ ID NO:99 respectively; (12) are SEQ ID NO:103, SEQ ID NO:104 and SEQ ID NO:105 respectively; (13) are SEQ ID NO:109, SEQ ID NO:110 and SEQ ID NO:111 respectively; (14) are SEQ ID NO:85, SEQ ID NO:110 and SEQ ID NO:117 respectively; (15) are SEQ ID NO:121, SEQ ID NO:122 and SEQ ID NO:123 respectively; (16) are SEQ ID NO:127, SEQ ID NO:128 and SEQ ID NO:129; (17) are SEQ ID NO:133, SEQ ID NO:134 and SEQ ID NO:135, respectively; (18) are SEQ ID NO:139, SEQ ID NO:140 and SEQ ID NO:141, respectively;(19) are SEQ ID NO:145, SEQ ID NO:146 and SEQ ID NO:147, respectively; (20) are SEQ ID NO:31, SEQ ID NO:32 and SEQ ID NO:33, respectively; (21) are SEQ ID NO:31, SEQ ID NO:32 and SEQ ID NO:159, respectively; (22) are SEQ ID NO:169, SEQ ID NO:170 and SEQ ID NO:171, respectively; (23) are SEQ ID NO:61, SEQ ID NO:62 and SEQ ID NO:63, respectively; (24) are SEQ ID NO:31, SEQ ID NO:182 and SEQ ID NO:183, respectively; (25) are SEQ ID NO:187, SEQ ID NO:188 and SEQ ID NO:189, respectively; (26) are SEQ ID NO:193, SEQ ID NO:194 and SEQ ID NO:195, respectively; (27) are SEQ ID NO:199, SEQ ID NO:200 and SEQ ID NO: 201; (28) SEQ ID NO: 205, SEQ ID NO: 206 and SEQ ID NO: 207, respectively; (29) SEQ ID NO: 211, 212 and 213, respectively; (30) SEQ ID NO: 127, 386 and 129, respectively; (31) SEQ ID NO: 205, 206 and 207, respectively; (32) SEQ ID NO: 127, 388 and 129, respectively; or (33) SEQ ID NO: 205, 387 and 207, respectively, wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO: ;

[0081] According to a variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, which comprises at least one VL CDR having an amino acid sequence as follows: (1) SEQ ID NO: 34, 35 or 36; (2) SEQ ID NO: 40, 41 or 42; (3) SEQ ID NO: 46, 47 or 48; (4) SEQ ID NO: 58, 59 or 60; (5) SEQ ID NO: 64, 65 or 66; (6) SEQ ID NO: 34, 35 or 72; (7) SEQ ID NO: 76, 47 or 78; (8) SEQ ID NO: 82, 83 or 84; (9) SEQ ID NO: 88, 89 or 90; (10) SEQ ID NO: 94, 95 or 96; (11) SEQ ID NO: 100, 101 or 102; (12) SEQ ID NO: 64, 107 or 108; (13) SEQ ID NO: NO:88, 89 or 114; (14) SEQ ID NO:124, 125 or 126; (15) SEQ ID NO:34, 35 or 36; (16) SEQ ID NO:136, 137 or 138; (17) SEQ ID NO:142, 143 or 144; (18) SEQ ID NO:148, 149 or 150; (19) SEQ ID NO:136, 137 or 162; (20) SEQ ID NO:34, 35 or 36; (21) SEQ ID NO:172, 173 or 174; (22) SEQ ID NO:64, 65 or 180; (23) SEQ ID NO:136, 137 or 186; (24) SEQ ID NO:190, 191 or 192; (25) SEQ ID NO:196, 197 or 12; (26) SEQ ID NO:202, 203 or 6; (27) SEQ ID NO:142, 209 or 210; or (28) SEQ ID NO:214, 35 or 216; (29) SEQ ID NO:10, 11 or 12; (30) SEQ ID NO:4, 5 or 6; or (31) SEQ ID NO:142, 143 or 210, wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO:.

[0082] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, which comprises a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2 and CDRL3, wherein CDRL1, CDRL2 and CDRL3 have the following amino acid sequences: (1) SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, respectively; (2) SEQ ID NO:40, SEQ ID NO:41 and SEQ ID NO:42, respectively; (3) SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48, respectively; (4) SEQ ID NO:58, SEQ ID NO:59 and SEQ ID NO:60, respectively; (5) SEQ ID NO:64, SEQ ID NO:65 and SEQ ID NO:66, respectively; (6) SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:72, respectively; (7) SEQ ID NO:76, SEQ ID NO:47 and SEQ ID NO:78, respectively. NO:78; (8) are SEQ ID NO:82, SEQ ID NO:83 and SEQ ID NO:84, respectively; (9) are SEQ ID NO:88, SEQ ID NO:89 and SEQ ID NO:90, respectively; (10) are SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96, respectively; (11) are SEQ ID NO:100, SEQ ID NO:101 and SEQ ID NO:102, respectively; (12) are SEQ ID NO:64, SEQ ID NO:107 and SEQ ID NO:108, respectively; (13) are SEQ ID NO:88, SEQ ID NO:89 and SEQ ID NO:114, respectively; (14) are SEQ ID NO:124, SEQ ID NO:125 and SEQ ID NO:126, respectively; (15) are SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, respectively; (16) are SEQ ID NO:136, SEQ ID NO:137 and SEQ ID NO:138, respectively. NO:138; (17) are SEQ ID NO:142, SEQ ID NO:143 and SEQ ID NO:144, respectively; (18) are SEQ ID NO:148, SEQ ID NO:149 and SEQ ID NO:150, respectively;(19) are SEQ ID NO:136, SEQ ID NO:137 and SEQ ID NO:162, respectively; (20) are SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, respectively; (21) are SEQ ID NO:172, SEQ ID NO:173 and SEQ ID NO:174, respectively; (22) are SEQ ID NO:64, SEQ ID NO:65 and SEQ ID NO:180, respectively; (23) are SEQ ID NO:136, SEQ ID NO:137 and SEQ ID NO:186, respectively; (24) are SEQ ID NO:190, SEQ ID NO:191 and SEQ ID NO:192, respectively; (25) are SEQ ID NO:196, SEQ ID NO:197 and SEQ ID NO:12, respectively; (26) are SEQ ID NO:202, SEQ ID NO:203 and SEQ ID NO:6, respectively; (27) are SEQ ID NO:142, SEQ ID NO:209 and SEQ ID NO: ID NO: 210; (28) SEQ ID NO: 214, SEQ ID NO: 35 and SEQ ID NO: 216, respectively; (29) SEQ ID NO: 10, 11 or 12, respectively; (30) SEQ ID NO: 4, 5 or 6, respectively; or (31) SEQ ID NO: 142, 143 or 210, respectively, wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO: ;

[0083] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein (1) CDRH1, CDRH2, and CDRH3 have the amino acid sequences set forth in SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences set forth in SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36, respectively; and (2) CDRH1, CDRH2, and CDRH3 have the amino acid sequences set forth in SEQ ID NO:37, SEQ ID NO:38, and SEQ ID NO:39, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences set forth in SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42, respectively. NO:42; (3) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:43, SEQ ID NO:44 and SEQ ID NO:45, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48, respectively; (4) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:43, SEQ ID NO:56 and SEQ ID NO:57, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:58, SEQ ID NO:59 and SEQ ID NO:60, respectively; (5) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:61, SEQ ID NO:62 and SEQ ID NO:63, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:64, SEQ ID NO:65 and SEQ ID NO:66, respectively. NO:66; (6) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:69, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:72, respectively;(7) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 73, SEQ ID NO: 74, and SEQ ID NO: 75, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 76, SEQ ID NO: 47, and SEQ ID NO: 78, respectively; (8) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 79, SEQ ID NO: 80, and SEQ ID NO: 81, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 82, SEQ ID NO: 83, and SEQ ID NO: 84, respectively; (9) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 85, SEQ ID NO: 86, and SEQ ID NO: 87, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 90, respectively. NO:90; (10) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:61, SEQ ID NO:92 and SEQ ID NO:93, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96, respectively; (11) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:97, SEQ ID NO:98 and SEQ ID NO:99, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:100, SEQ ID NO:101 and SEQ ID NO:102, respectively; (12) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:103, SEQ ID NO:104 and SEQ ID NO:105, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:64, SEQ ID NO: The amino acid sequences shown in SEQ ID NO:107 and SEQ ID NO:108;(13) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 109, SEQ ID NO: 110, and SEQ ID NO: 111, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 114, respectively; (14) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 85, SEQ ID NO: 110, and SEQ ID NO: 117, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 114, respectively; (15) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 124, SEQ ID NO: 125, and SEQ ID NO: 126, respectively. (16) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively; (17) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 133, SEQ ID NO: 134, and SEQ ID NO: 135, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138, respectively; (18) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO: 141, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 143, and SEQ ID NO: 144, respectively. The amino acid sequences shown in SEQ ID NO:142, SEQ ID NO:143, and SEQ ID NO:144;(19) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 145, SEQ ID NO: 146, and SEQ ID NO: 147, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 148, SEQ ID NO: 149, and SEQ ID NO: 150, respectively; (20) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 148, SEQ ID NO: 149, and SEQ ID NO: 150, respectively; (21) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 159, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138, respectively. (22) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 169, SEQ ID NO: 170, and SEQ ID NO: 171, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 172, SEQ ID NO: 173, and SEQ ID NO: 174, respectively; (23) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 61, SEQ ID NO: 62, and SEQ ID NO: 63, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 64, SEQ ID NO: 65, and SEQ ID NO: 180, respectively; (24) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 182, and SEQ ID NO: 183, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 182, and SEQ ID NO: 183, respectively. the amino acid sequences shown in SEQ ID NO:136, SEQ ID NO:137, and SEQ ID NO:186;(25) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188, and SEQ ID NO: 189, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 190, SEQ ID NO: 191, and SEQ ID NO: 192, respectively; (26) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 193, SEQ ID NO: 194, and SEQ ID NO: 195, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 196, SEQ ID NO: 197, and SEQ ID NO: 12, respectively; (27) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO: 201, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: NO:203 and SEQ ID NO:6; (28) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:205, SEQ ID NO:206 and SEQ ID NO:207, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:142, SEQ ID NO:209 and SEQ ID NO:210, respectively; (29) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:211, SEQ ID NO:212 and SEQ ID NO:213, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:214, SEQ ID NO:35 and SEQ ID NO:216, respectively; (30) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:127, SEQ ID NO:386 and SEQ ID NO:387, respectively. NO:129, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36, respectively;(31) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 205, SEQ ID NO: 206, and SEQ ID NO: 207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 209, and SEQ ID NO: 210, respectively; (32) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 388, and SEQ ID NO: 129, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively; (33) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 205, SEQ ID NO: 387, and SEQ ID NO: 207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 388, and SEQ ID NO: 36, respectively. NO:209 and SEQ ID NO:210; or (34) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:205, SEQ ID NO:387, and SEQ ID NO:207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:142, SEQ ID NO:143, and SEQ ID NO:210, respectively; wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO:. ;

[0084] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising: (1) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO:31, SEQ ID NO:32, or SEQ ID NO:33, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO:34, SEQ ID NO:35, or SEQ ID NO:36; (2) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO:37, SEQ ID NO:38, or SEQ ID NO:39, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO:40, SEQ ID NO:41, or SEQ ID NO:42; (3) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO:43, SEQ ID NO:44, or SEQ ID NO:45, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NO:48; (4) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO:43, SEQ ID NO:44, or SEQ ID NO:45, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO:46, SEQ ID NO:47, or SEQ ID NO:48; NO:56 or SEQ ID NO:57, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO:58, SEQ ID NO:59 or SEQ ID NO:60; (5) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO:61, SEQ ID NO:62 or SEQ ID NO:63, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO:64, SEQ ID NO:65 or SEQ ID NO:66; (6) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO:31, SEQ ID NO:32 or SEQ ID NO:69, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO:34, SEQ ID NO:35 or SEQ ID NO:72; (7) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO:73, SEQ ID NO:74 or SEQ ID NO:75, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO:76, SEQ ID NO:77 or SEQ ID NO:78; VL CDR of the amino acid sequence shown in NO:78;(8) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 79, SEQ ID NO: 80 or SEQ ID NO: 81, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 82, SEQ ID NO: 83 or SEQ ID NO: 84; (9) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 88, SEQ ID NO: 89 or SEQ ID NO: 90; (10) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 61, SEQ ID NO: 92 or SEQ ID NO: 93, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 94, SEQ ID NO: 95 or SEQ ID NO: 96; (11) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 97, SEQ ID NO: 98 or SEQ ID NO: 99, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: : (14) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 85, SEQ ID NO: 110 or SEQ ID NO: 117, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 88, SEQ ID NO: 89 or SEQ ID NO: 114; (15) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 85, SEQ ID NO: 110 or SEQ ID NO: 117, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 88, SEQ ID NO: 89 or SEQ ID NO: 114. CDR;(15) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 121, SEQ ID NO: 122 or SEQ ID NO: 123, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 124, SEQ ID NO: 125 or SEQ ID NO: 126; (16) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 127, SEQ ID NO: 128 or SEQ ID NO: 129, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO: 36; (17) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 133, SEQ ID NO: 134 or SEQ ID NO: 135, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 136, SEQ ID NO: 137 or SEQ ID NO: 138; (18) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 139, SEQ ID NO: 140 or SEQ ID NO: : (19) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 145, SEQ ID NO: 146 or SEQ ID NO: 147, and at least one VLCDR having an amino acid sequence as shown in SEQ ID NO: 148, SEQ ID NO: 149 or SEQ ID NO: 150; (20) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 33, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 148, SEQ ID NO: 149 or SEQ ID NO: 150; (21) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 159, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 160; VL CDRs of the amino acid sequence shown in SEQ ID NO: 136, SEQ ID NO: 137, or SEQ ID NO: 162;(22) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 169, SEQ ID NO: 170 or SEQ ID NO: 171, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 172, SEQ ID NO: 173 or SEQ ID NO: 174; (23) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 180; (24) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 31, SEQ ID NO: 182 or SEQ ID NO: 183, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 136, SEQ ID NO: 137 or SEQ ID NO: 186; (25) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 187, SEQ ID NO: 188 or SEQ ID NO: NO: 189, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 190, SEQ ID NO: 191 or SEQ ID NO: 192; (26) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 193, SEQ ID NO: 194 or SEQ ID NO: 195, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 12; (27) at least one VH CDR having an amino acid sequence as shown in SEQ ID NO: 199, SEQ ID NO: 200 or SEQ ID NO: 201, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: 202, SEQ ID NO: 203 or SEQ ID NO: 6; (28) at least one VHCDR having an amino acid sequence as shown in SEQ ID NO: 205, SEQ ID NO: 206 or SEQ ID NO: 207, and at least one VL CDR having an amino acid sequence as shown in SEQ ID NO: NO: 142, SEQ ID NO: 209 or SEQ ID NO: 210;(29) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 211, SEQ ID NO: 212 or SEQ ID NO: 213, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 214, SEQ ID NO: 35 or SEQ ID NO: 216; (30) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 127, SEQ ID NO: 386 or SEQ ID NO: 129, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO: 36; (31) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 205, SEQ ID NO: 206 or SEQ ID NO: 207, and at least one VL CDR having an amino acid sequence as set forth in SEQ ID NO: 142, SEQ ID NO: 209 or SEQ ID NO: 210; (32) at least one VH CDR having an amino acid sequence as set forth in SEQ ID NO: 127, SEQ ID NO: 388 or SEQ ID NO: NO: 129, and at least one VL CDR having the amino acid sequence of SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO: 36; (33) at least one VH CDR having the amino acid sequence of SEQ ID NO: 205, SEQ ID NO: 387 or SEQ ID NO: 207, and at least one VL CDR having the amino acid sequence of SEQ ID NO: 142, SEQ ID NO: 209 or SEQ ID NO: 210; or (34) at least one VH CDR having the amino acid sequence of SEQ ID NO: 205, SEQ ID NO: 387 or SEQ ID NO: 207, and at least one VL CDR having the amino acid sequence of SEQ ID NO: 142, SEQ ID NO: 143 or SEQ ID NO: 210; wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO: .;

[0085] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 13, or a sequence having at least 90% sequence identity thereto. In other embodiments, the heavy chain variable region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO: 17 or 21.

[0086] For any embodiment disclosed herein involving at least 90% sequence identity, it is understood that this includes any sequence identity between 90% and 100%, i.e., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and 100%.

[0087] In other embodiments, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:301, 302, 303, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 324, 306, 326, 327, 330, 331, 13, 17, 21, 382, ​​384, 389, 390 and 378.

[0088] In other embodiments, the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 301, 302, 303, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 324, 306, 326, 327, 330, 331, 13, 17, 21, 382, ​​384, 389, 390 and 378, wherein there are up to 10 modifications, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid modifications.

[0089] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the light chain comprises a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 14, or a sequence having at least 90% sequence identity thereto. In other embodiments, the light chain variable region comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO: 15 or 19.

[0090] In other embodiments, the light chain variable region comprises an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:351, 352, 353, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 366, 367, 368, 369, 370, 372, 351, 374, 375, 376, 377, 380, 381, 14, 15, 19, 383, 385 or 378.

[0091] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 13, and the light chain comprises a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 14.

[0092] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, which comprises a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 13, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 14.

[0093] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, which comprises a heavy chain and a light chain, wherein: (1) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:17, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:15; or (2) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:21, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:19.

[0094] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA is provided, comprising a heavy chain and a light chain, wherein: (1) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 301, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 351; (2) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 302, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 352; (3) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 303, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: NO: 353; (4) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 305, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 355; (5) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 306, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 356; (6) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 307, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 308. (7) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 308, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 358;(8) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 309, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 359; (9) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 310, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 360; (10) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 311, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 312. NO: 361; (11) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 312, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 362; (12) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 313, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 363; (13) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 314, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 315. NO:364; (14) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:315, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:364;(15) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 316, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 366; (16) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 317, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 367; (17) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 318, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 319. NO: 368; (18) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 319, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 369; (19) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 320, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 370; (20) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 321, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 322. NO:370; (21) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:322, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:372;(22) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 324, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 374; (23) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 306, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 375; (24) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 326, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 327. NO:376; (25) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:327, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:377; (26) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:17, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:15; (27) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:21, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:23. NO:19; (28) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:330, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:380;(29) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 331, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 381; (30) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 382, ​​and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 383; (31) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 384, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 385. NO: 385; (32) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 389, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 383; (33) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 390, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 385; or (34) the heavy chain comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 390, and the light chain comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: An amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in NO:378.

[0095] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:13, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:14.

[0096] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:17, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:15.

[0097] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:21, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:19.

[0098] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:301, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:351.

[0099] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:302, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:352.

[0100] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:303, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:353.

[0101] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:305, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:355.

[0102] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:306, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:356.

[0103] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:307, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:357.

[0104] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:308, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:358.

[0105] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:309, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:359.

[0106] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:310, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:360.

[0107] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:311, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:361.

[0108] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:312, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:362.

[0109] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:313, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:363.

[0110] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:314, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:364.

[0111] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:315, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:364.

[0112] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:316, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:366.

[0113] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:317, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:367.

[0114] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:318, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:368.

[0115] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:319, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:369.

[0116] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:320, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:370.

[0117] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:321, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:370.

[0118] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:322, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:372.

[0119] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:324, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:374.

[0120] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:306, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:375.

[0121] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:326, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:376.

[0122] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:327, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:377.

[0123] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:17, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:15.

[0124] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:21, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:19.

[0125] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:330, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:380.

[0126] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:331, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:381.

[0127] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:382, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:383.

[0128] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:384, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:385.

[0129] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:389, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:383.

[0130] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:390, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:385.

[0131] In one embodiment, the heavy chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:390, and the light chain variable region comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:378.

[0132] In other embodiments, the heavy chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:13.

[0133] In other embodiments, the heavy chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:17.

[0134] In other embodiments, the heavy chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:21.

[0135] In other embodiments, the heavy chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:301, 302, 303, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 324, 306, 326, 327, 330, 331, 382, ​​384, 389 or 390.

[0136] In other embodiments, the light chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:14.

[0137] In other embodiments, the light chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:15.

[0138] In other embodiments, the light chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:19.

[0139] In other embodiments, the light chain variable region polypeptide is at least 92%, at least 95%, at least 97%, at least 98% or at least 99% identical to the sequence disclosed in SEQ ID NO:351, 352, 353, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 364, 366, 367, 368, 369, 370, 372, 374, 375, 376, 377, 380, 381, 383, 385 or 378.

[0140] According to another variation of the first aspect of the present invention, an isolated antibody or antigen-binding fragment thereof is provided, which has a primary VH domain, a primary VL domain, CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 of any antibody clone shown in Table 1. In certain embodiments, provided herein is an isolated antibody selected from the antibody clones shown in Table 1.

[0141] Table 1. Exemplary BTLA agonistic antibodies

[0142]

[0143]

[0144] In certain embodiments, the heavy or light chain further comprises a constant region.If the molecule is a full-length IgG-type antibody molecule, the heavy chain may comprise three constant domains.

[0145] In certain embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA exhibits at most about 10 x 10 -9 M's K D In certain embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA exhibits at most about 4×10 -9 M's K D In certain embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to human BTLA exhibits at most about 1×10 -9 M's K D .

[0146] In certain embodiments, the isolated antibodies (e.g., humanized) of the invention are expressed at a concentration of no more than about 10 nM (1 x 10 -8 M)K D Binds to human BTLA; suitably no more than about 1 nM; more suitably wherein the antibody has a K at 37°C D The value does not exceed about 500 pM (5x10 -10The term "about" as used herein refers to + / - 10%.

[0147] In certain embodiments, the isolated antibodies (e.g., humanized) of the invention have a pH of at least 1.0 x 10 5 In certain embodiments, the isolated antibodies (e.g., humanized) of the present invention bind to human BTLA at an on-rate of at least 2.0 x 10 5 (1 / Ms), 3.0x10 5 (1 / Ms), 4.0x10 5 (1 / Ms), 5.0x10 5 (1 / Ms), 6.0x10 5 (1 / Ms) or 7.0x10 5 The on-rate of 1 / Ms binds to human BTLA.

[0148] In certain embodiments, the isolated antibodies (e.g., humanized) of the present invention have a pH of no more than or less than 1.0 x 10 -3 In certain embodiments, the isolated antibodies (e.g., humanized) of the present invention bind to human BTLA at an off-rate of no more than or less than 3.0 x 10 -4 In certain embodiments, the isolated antibodies (e.g., humanized) of the present invention bind to human BTLA at an off-rate of no more than or less than 2.0 x 10 -4 (1 / s) or 1.0x10 -4 The off-rate of 1 / s binds to human BTLA.

[0149] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10 nM as determined by surface plasmon resonance (SPR) at 37° C. using, for example, the methods described in Example 2, and wherein the antibody binds to cynomolgus monkey BTLA with a KD of less than 20 nM as determined by surface plasmon resonance (SPR) at 37° C. using, for example, the methods described in Example 2; does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits the proliferation of T cells in vitro as determined by, for example, a mixed lymphocyte reaction assay using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has a KD of at least 5.0 x 10 5 In some embodiments, the antibody or antigen-binding fragment thereof binds to human B and T lymphocyte attenuator (BTLA) at an on-rate of less than 3.0 x 10 -4 In some embodiments, the antibody or antigen-binding fragment thereof has an off-rate of 3.0 x 10 -4 (1 / s) to 1.0x10 -3 (1 / s) dissociation rate binds to human B and T lymphocyte attenuator (BTLA). In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106 and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using the methods described in Example 5. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from Y39, K41, R42, Q43, E45 and S47. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from D35, T78, K81, S121 and L123. In some embodiments, the antibody or its antigen-binding fragment binds to residue H68 of human BTLA. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from N65 and A64.

[0150] Methods for characterizing the properties of the antibodies or antigen-binding fragments thereof of the present invention are well known in the art. A suitable method for determining binding specificity using surface plasmon resonance (SPR) at 37°C is described in Example 2. A suitable method for determining whether a tested antibody / fragment thereof inhibits BTLA binding to herpes virus entry mediator (HVEM) is described in Example 4; this also employs surface plasmon resonance (SPR). A suitable method for determining whether a tested antibody / fragment thereof inhibits T cell proliferation in vitro is a mixed lymphocyte reaction assay, such as the assay described in Example 9. A suitable method for determining the binding site of an antibody / fragment thereof to BTLA may utilize X-ray crystallography or flow cytometry of mutant receptors, such as by the methods described in Example 5.

[0151] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof having an affinity for at least 5.0 x 10 5 The antibody specifically binds to human B and T lymphocyte attenuator (BTLA) with an on-rate of (1 / Ms), wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined, for example, by surface plasmon resonance (SPR), using the methods described, for example, in Example 4; and wherein the antibody inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay, using the methods described, for example, in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has an on-rate of less than 3.0 x 10 -4(1 / s) dissociation rate binds to human B and T lymphocyte attenuator (BTLA). In some embodiments, the antibody or its antigen-binding fragment binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in Example 2, for example. In some embodiments, the antibody or its antigen-binding fragment binds to cynomolgus monkey BTLA with a KD of less than 20nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in Example 2, for example. In some embodiments, the antibody or its antigen-binding fragment binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92 as determined by X-ray crystallography of mutant receptors or by flow cytometry using the method described in Example 5, for example. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0152] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof having an affinity for 3.0 x 10 -4 (1 / s) to 1.0x10 -3(1 / s) and specifically binds to human B and T lymphocyte attenuator (BTLA) with an off-rate of 1 / s, wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined, for example, by surface plasmon resonance (SPR) using the methods described in Example 4; and wherein the antibody inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay using the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10 nM as determined by surface plasmon resonance (SPR) at 37°C using the methods described in Example 2. In some embodiments, the antibody or antigen-binding fragment thereof binds to cynomolgus monkey B and T lymphocyte attenuator (BTLA) with a KD of less than 20 nM as determined by surface plasmon resonance (SPR) at 37°C using the methods described in Example 2, for example. In some embodiments, the antibody or antigen-binding fragment thereof has an affinity for at least 5.0 x 10 5 (1 / Ms) of the association rate binds to human B and T lymphocyte attenuator (BTLA). In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106 and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using the methods described in Example 5. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from Y39, K41, R42, Q43, E45 and S47. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from D35, T78, K81, S121 and L123. In some embodiments, the antibody or its antigen-binding fragment binds to residue H68 of human BTLA. In some embodiments, the antibody or its antigen-binding fragment binds to residues of human BTLA selected from N65 and A64.

[0153] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof, each having a surface plasmon resonance (SPR) density of less than 1.0 x 10 -3 (1 / s) and a dissociation rate of at least 5.0x10 5(1 / Ms) specifically binds to human B and T lymphocyte attenuator (BTLA), wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined, for example, by surface plasmon resonance (SPR) using the methods described in Example 4; and wherein the antibody inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay using the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10 nM as determined by surface plasmon resonance (SPR) at 37°C using the methods described in Example 2. In some embodiments, the antibody or antigen-binding fragment thereof binds to cynomolgus monkey BTLA with a KD of less than 20 nM as determined by surface plasmon resonance (SPR) at 37°C using the methods described in Example 2, for example. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using the methods described in, for example, Example 5. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0154] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA) with a KD of less than 2 nM as determined by surface plasmon resonance (SPR) at 37° C. using, for example, the methods described in Example 2, wherein the antibody inhibits the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits the proliferation of T cells in vitro, e.g., by a mixed lymphocyte reaction assay, as determined using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has a KD of less than 1.0 x 10 6In some embodiments, the antibody or antigen-binding fragment thereof binds to human B and T lymphocyte attenuator (BTLA) at an on-rate of less than 1.0 x 10 -3 (1 / s) of the dissociation rate binds to human B and T lymphocyte attenuator (BTLA). In some embodiments, the antibody or its antigen-binding fragment binds to cynomolgus monkey B and T lymphocyte attenuator (BTLA) with a KD of less than 10nM as determined by surface plasmon resonance (SPR) at 37°C using, for example, the method described in Example 2. In some embodiments, the antibody or its antigen-binding fragment binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using, for example, the method described in Example 5. In some embodiments, the antibody or its antigen-binding fragment binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or its antigen-binding fragment binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0155] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof having a specific affinity for agonist antibodies as determined by surface plasmon resonance (SPR) at 37°C of less than 1 x 10 -3(1 / s) dissociation rate specifically binds to human B and T lymphocyte attenuator (BTLA), wherein the antibody inhibits the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using the method described in Example 4, for example; and inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay using the method described in Example 9. In some embodiments, the antibody or its antigen-binding fragment binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10 nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in Example 2, for example. In some embodiments, the antibody or its antigen-binding fragment binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 2 nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in Example 2, for example. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using the methods described in, for example, Example 5. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0156] In certain embodiments of the first aspect of the invention, provided herein are isolated agonistic antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA), wherein the antibody binds to cynomolgus monkey BTLA with a KD of at least 5 nM as determined by surface plasmon resonance (SPR) at 37° C. using, for example, the methods described in Example 2; and wherein the antibody inhibits the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits the proliferation of T cells in vitro, e.g., as determined by a mixed lymphocyte reaction assay using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using, for example, the methods described in Example 5. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0157] In certain embodiments of the first aspect of the invention, provided herein are isolated agonistic antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA), wherein the antibody binds to cynomolgus monkey BTLA with a KD of at least 50 nM as determined by surface plasmon resonance (SPR) at 37° C. using, for example, the methods described in Example 2; and wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits T cell proliferation in vitro as determined, for example, by a mixed lymphocyte reaction assay using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92 as determined by X-ray crystallography of mutant receptors or by flow cytometry using, for example, the methods described in Example 5. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0158] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA) with a KD of 1400 nM to 3500 nM as determined by surface plasmon resonance (SPR) at 37°C using, for example, the methods described in Example 2; and wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has a KD of at least 2.0 x 10 5 In some embodiments, the antibody or antigen-binding fragment thereof binds to human BTLA at an on-rate of less than 10.0 x 10-1 (1 / s) off-rate binds to human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of the mutant receptor or by flow cytometry using the methods described in Example 5, for example. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0159] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof having an affinity for 1.7 x 10 5 (1 / Ms) to 2.5x10 5 (1 / Ms) of the association rate of human B and T lymphocyte attenuator (BTLA); and wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using the method described, for example, in Example 4; and inhibits the proliferation of T cells in vitro as determined, for example, by a mixed lymphocyte reaction assay using the method described, for example, in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has an on-off rate of less than 3.0 x 10 -1 In some embodiments, the antibody or antigen-binding fragment thereof binds to human BTLA at an off-rate of 3.0 x 10 -1 (1 / s) to 5.0x10 -1(1 / s) dissociation rate binds to human BTLA. In some embodiments, the antibody or its antigen-binding fragment binds to human BTLA with a KD of at least 150nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in, for example, Example 2. In some embodiments, the antibody or its antigen-binding fragment binds to human BTLA with a KD of 150nM to 1500nM as determined by surface plasmon resonance (SPR) at 37°C using the method described in, for example, Example 2. In some embodiments, the antibody or its antigen-binding fragment binds to an epitope that blocks the binding of the 286 antibody. In some embodiments, the antibody or its antigen-binding fragment binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of mutant receptors or by flow cytometry using the method described in, for example, Example 5. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0160] In certain embodiments of the first aspect of the invention, provided herein are isolated agonist antibodies or antigen-binding fragments thereof that specifically bind to human B and T lymphocyte attenuator (BTLA) with a KD of 40 nM to 1200 nM as determined by surface plasmon resonance (SPR) at 37°C using, for example, the methods described in Example 2; and wherein the antibody does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM) as determined by surface plasmon resonance (SPR) using, for example, the methods described in Example 4; and inhibits the proliferation of T cells in vitro, for example, as determined by a mixed lymphocyte reaction assay using, for example, the methods described in Example 9. In some embodiments, the antibody or antigen-binding fragment thereof has a KD of at least 1.0 x 10 5 In some embodiments, the antibody or antigen-binding fragment thereof binds to human BTLA at an on-rate of 1.0 x 10 5 (1 / Ms) to 10x10 5In some embodiments, the antibody or antigen-binding fragment thereof binds to human BTLA at an on-rate of less than 6.0 x 10 -1 In some embodiments, the antibody or antigen-binding fragment thereof binds to human BTLA at an off-rate of 6.0 x 10 -1 (1 / s) to 10.0x10 -2 (1 / s) off-rate binds to human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92, as determined by X-ray crystallography of the mutant receptor or by flow cytometry using the methods described in Example 5, for example. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from Y39, K41, R42, Q43, E45, and S47. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from D35, T78, K81, S121, and L123. In some embodiments, the antibody or antigen-binding fragment thereof binds to residue H68 of human BTLA. In some embodiments, the antibody or antigen-binding fragment thereof binds to a residue of human BTLA selected from N65 and A64.

[0161] In certain embodiments, an isolated antibody or antigen-binding fragment thereof of the invention that specifically binds to human BTLA increases BTLA activity and / or signaling through the receptor.

[0162] In certain embodiments, the antibody of the present invention is selected from the group consisting of: a human antibody, a humanized antibody, a chimeric antibody, and a multispecific antibody (eg, a bispecific antibody).

[0163] In certain embodiments, the antigen-binding fragment of the invention is selected from the group consisting of: scFv, sc(Fv) 2 , dsFv, Fab, Fab', (Fab')2 and diabodies.

[0164] In certain embodiments, the heavy chain and light chain molecules forming the antigen binding fragment are connected by a flexible linker. There are many commonly used flexible linkers, and the selection of the linker can be made by those skilled in the art.

[0165] The peptide connector connecting the scFv VH and VL domains connects the carboxyl terminus of one variable region domain to the amino terminus of the other variable domain without significantly compromising the fidelity of VH-VL pairing and antigen binding site. The length of the peptide connector can vary from 10 to 25 amino acids and is usually, but not always, composed of hydrophilic amino acids such as glycine (G) and serine (S). The connector can be a connector found in natural multidomain proteins (e.g., see Argos PJ Mol Biol. 211: 943-958, 1990; and Heringa G. Protein Eng. 15: 871-879, 2002), or a modified form thereof.

[0166] Commonly used flexible linkers have a sequence consisting primarily of a stretch of Gly and Ser residues ("GS" linker). An example of the most widely used flexible linker is the sequence (Gly-Gly-Gly-Gly-Ser) n . By adjusting the copy number "n", the length of the GS linker can be changed to achieve appropriate separation of functional domains or to maintain necessary interdomain interactions. Typically, the (GGGGS)3 peptide is used as a scFv peptide linker (Leith et al., Int. J. Oncol. 24:765–771, 2004; Holger et al. Proc. Natl. Acad. Sci. USA 90:6444–6448, 1993). This 15-amino acid linker sequence [called the (GGGGS)3 linker] is used in the recombinant phage antibody system (RPAS kit) commercially available from Amersham. Several other linkers have also been used to create scFV molecules (e.g., KESGSVSSEQLAQFRSLD and EGKSSGSGSESKST; Bird et al., Science 242:432-426, 1988).

[0167] The inventors have mapped the epitope on BTLA to which the potent 2.8.6 and 11.5.1 agonists and other antibodies disclosed herein bind.

[0168] In a specific embodiment, an antibody or antigen-binding fragment thereof of the invention binds to a residue of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, E92, Y39, K41, R42, Q43, E45, S47, D35, T78, K81, S121, L123, H68, N65, A64.

[0169] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to a residue of human BTLA selected from the group consisting of D52, P53, E55, E57, E83, Q86, E103, L106, E92.

[0170] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to at least two residues of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, E92.

[0171] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to at least three residues of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92.

[0172] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to at least five residues of human BTLA selected from D52, P53, E55, E57, E83, Q86, E103, L106, and E92.

[0173] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to all residues of human BTLA selected from the group consisting of D52, P53, E55, E57, E83, Q86, E103, L106, and E92.

[0174] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to a residue of human BTLA selected from the group consisting of Y39, K41, R42, Q43, E45, and S47.

[0175] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to at least two residues of human BTLA selected from Y39, K41, R42, Q43, E45, and S47.

[0176] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to all residues of human BTLA selected from the group consisting of Y39, K41, R42, Q43, E45, and S47.

[0177] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to a residue of human BTLA selected from the group consisting of D35, T78, K81, S121, and L123.

[0178] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to at least two residues of human BTLA selected from D35, T78, K81, S121, and L123.

[0179] In a specific embodiment, an antibody or antigen-binding fragment thereof of the invention binds to residue H68 of human BTLA.

[0180] In certain embodiments, an antibody or antigen-binding fragment thereof of the invention binds to a residue of human BTLA selected from the group consisting of N65 and A64.

[0181] In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention bind to both N65 and A64 residues of human BTLA.

[0182] Residue numbering, such as K41, refers to the amino acid at position 41 (K; lysine); wherein the numbering refers to the position in the human BTLA polypeptide disclosed in SEQ ID NO:23.

[0183] In certain embodiments, the antibodies of the invention are IgG1, IgG2, or IgG4 antibodies. In certain embodiments, the antibodies are murine or human antibodies.

[0184] In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention are humanized antibodies.

[0185] In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention are fully human antibodies.

[0186] In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention act as agonists that induce signaling through the BTLA receptor.

[0187] The antibodies (or antigen-binding fragments thereof) of the invention are particularly potent agonists.

[0188] In certain embodiments, the EC50 of an antibody or antigen-binding fragment thereof of the invention is no greater than 1 nM.

[0189] The agonist antibodies (or antigen-binding fragments thereof) of the present invention have particularly high efficacy.

[0190] In particular embodiments, the antibodies or antigen-binding fragments thereof of the invention inhibit T cell proliferation by at least 20%, suitably at least 30%, more suitably at least 40%.

[0191] In a particular embodiment, the antibodies or antigen-binding fragments thereof of the invention inhibit T cell IFN-γ production by at least 50%, suitably at least 75%, more suitably at least 95%, as measured, for example, by ELISA of supernatants from an in vitro mixed lymphocyte reaction.

[0192] In a particular embodiment, the antibodies or antigen-binding fragments thereof of the invention inhibit T cell IL-2 production by at least 50%, suitably at least 75%, more suitably at least 95%, as measured, for example, by ELISA of supernatants from an in vitro mixed lymphocyte reaction.

[0193] In a particular embodiment, the antibodies or antigen-binding fragments thereof of the invention inhibit T cell IL-17 production by at least 50%, suitably at least 75%, more suitably at least 95%, as measured, for example, by ELISA of supernatants from an in vitro mixed lymphocyte reaction.

[0194] In a specific embodiment, using the methods as described in Example 12, the antibodies or antigen-binding fragments thereof of the invention reduce mortality in a murine GVHD model by at least 50%, suitably at least 75%, more suitably at least 95%.

[0195] In a specific embodiment, the antibodies or antigen-binding fragments thereof of the invention reduce body weight loss in a murine T-cell colitis model by at least 50%, suitably at least 75%, more suitably at least 95%, using the methods as described in Example 11.

[0196] In a specific embodiment, the antibodies or antigen-binding fragments thereof of the invention reduce colonic inflammation in a murine T-cell colitis model by at least 50%, suitably at least 75%, more suitably at least 95%, using the methods as described in Example 11.

[0197] In certain aspects, the present invention also relates to isolated polypeptides comprising the VL domain or VH domain of any of the antibodies or antigen-binding fragments thereof described herein.

[0198] In certain embodiments, the isolated polypeptide comprises the amino acid sequence shown in SEQ ID NO: 13 or 14, or a sequence at least 90% identical thereto.

[0199] Nucleic acid molecules

[0200] The antibody of the present invention or its antigen-binding fragment will be by nucleic acid encoding.This antibody or its antigen-binding fragment can be encoded by single nucleic acid molecule, or it can be encoded by two or more nucleic acid molecules.For example, because antigen binding site is usually formed by heavy chain variable polypeptide region and light chain variable polypeptide region combination together, these two variable (heavy chain and light chain) polypeptide regions can be encoded by independent nucleic acid molecules.Or, for example, in the case of ScFv, they can be encoded by identical nucleic acid molecules.

[0201] According to a second aspect of the present invention, there is provided one or more nucleic acid molecules encoding the antibody or antigen-binding fragment thereof according to the first aspect of the present invention.

[0202] A person skilled in the art is able to determine the appropriate nucleotide sequence encoding the polypeptide from the primary amino acid sequence of the polypeptide encoding the antibody or antigen-binding fragment thereof of the present invention, and, if necessary, determine the codon-optimized sequence (see, for example, Mauro and Chappell. Trends Mol Med. 20(11):604-613, 2014).

[0203] As used herein, when reference is made to the aforementioned aspects of the invention, for example "according to the first (or second, etc.) aspect of the invention", it should be understood to also cover any listed variations of said aspects (for example variations of the first (or second, etc.) aspect).

[0204] According to a variation of the second aspect of the present invention, an isolated nucleic acid is provided, comprising a nucleotide sequence encoding a heavy chain variable region polypeptide or a light chain variable region polypeptide of the present invention. The heavy chain variable polypeptide or light chain variable polypeptide of the present invention refers to a single polypeptide chain comprising amino acids that constitute a portion of an antigen binding site. Of course, the polypeptide may also comprise other domains, such as a constant domain, a hinge region, and an Fc region, for example, a region comprising one or more Fc receptor binding sites.

[0205] According to another variation of the second aspect of the present invention, an isolated nucleic acid is provided comprising one or more nucleotide sequences encoding a polypeptide capable of forming an antibody or antigen-binding fragment of the present invention. In certain embodiments, the polypeptide may further comprise other domains, such as a constant domain, a hinge region, and an Fc region, e.g., a region comprising one or more Fc receptor binding sites.

[0206] One of the nucleic acid molecules may encode only a polypeptide comprising the VL domain of an antibody or fragment thereof. One of the nucleic acid molecules may encode only a polypeptide comprising the VH domain of an antibody or fragment thereof. However, the nucleic acid molecule may also encode both VH and VL domains comprising a polypeptide sequence capable of forming an antibody or antibody fragment thereof of the invention.

[0207] The nucleic acid molecules encoding the antibodies or antigen-binding fragments thereof of the present invention, for example, according to the first aspect of the present invention, can be a vector (such as a plasmid vector, a cosmid vector or a viral vector, or an artificial chromosome) or can be part of such a vector, which can contain other functional regions (elements), such as one or more promoters, one or more replication origins, one or more selection markers, and one or more other elements commonly found in expression vectors. The cloning and expression of nucleic acids encoding proteins (including antibodies) are well established and within the skill of those skilled in the art.

[0208] According to a third aspect of the present invention, there is provided a vector comprising the nucleic acid of the second aspect of the present invention. In a particular embodiment, the vector is a plasmid vector, a cosmid vector, a viral vector or an artificial chromosome.

[0209] Nucleic acids of the invention, including vector nucleic acids comprising nucleotide sequences encoding polypeptides capable of forming antibodies or antigen-binding fragments thereof of the invention, can be in purified / isolated form.

[0210] Isolated / purified nucleic acids encoding antibodies of the invention or antigen-binding fragments thereof will be free or substantially free of materials with which they are naturally associated, such as other proteins or nucleic acids found in their natural environment or in the environment in which they are produced, such as cell culture, when such production is performed by recombinant DNA techniques performed in vitro or in vivo.

[0211] In particular embodiments, the nucleic acids of the invention are greater than 80%, such as greater than 90%, greater than 95%, greater than 97%, and greater than 99% pure.

[0212] Therefore, according to another variation of the third aspect of the present invention, a vector comprising a nucleic acid or nucleotide sequence encoding a heavy chain variable polypeptide or light chain variable polypeptide of the present invention is provided. In a specific embodiment, the vector comprises nucleic acids encoding the heavy chain and light chain variable regions. In a specific embodiment, the polypeptide may further comprise other domains, such as a constant domain, a hinge region, and an Fc region, for example, a region comprising one or more Fc receptor binding sites.

[0213] Nucleic acid of the present invention and / or vector can be introduced into host cell.This introduction can adopt any available technology.For eukaryotic cells, suitable technology can comprise calcium phosphate transfection, DEAE-dextran, electroporation, liposome-mediated transfection, and use retrovirus or other virus such as vaccinia virus or use baculovirus transduction for insect cells.In host cell, particularly eukaryotic cell, introduce nucleic acid and can use viral system or plasmid-based system.Plasmid system can be kept in episomal form, or can be incorporated in host cell or artificial chromosome.Incorporation can be completed by integrating one or more copies at random or targeting at single or multiple loci.For bacterial cells, suitable technology can comprise calcium chloride transformation, electroporation and use phage transfection.

[0214] In one embodiment, the nucleic acid of the invention is integrated into the genome (eg, chromosome) of the host cell. Integration can be facilitated by including sequences that promote recombination with the genome according to standard techniques.

[0215] host cells

[0216] Another aspect of the present invention provides a host cell comprising a nucleic acid disclosed herein.Such a host cell can be in vitro and can be in culture.

[0217] The host cell may be from any species, such as bacteria or yeast, but suitably the host cell is a mammalian cell, such as a human cell or a rodent cell, for example a HEK293T cell or a CHO-K1 cell.

[0218] Therefore, according to a fourth aspect of the present invention, there is provided a host cell comprising the nucleic acid sequence according to the second aspect of the present invention or the vector according to the third aspect of the present invention.

[0219] Host cells may be treated to cause or allow expression of a protein of the invention from a nucleic acid, for example by culturing the host cells under conditions for expression of the encoding nucleic acid. Purification of the expression product may be achieved by methods known to those skilled in the art.

[0220] Therefore, nucleic acids of the present invention, including vector nucleic acids comprising nucleotide sequences encoding polypeptides capable of forming antibodies of the present invention or antigen-binding fragments thereof, can be present in isolated host cells. Host cells are typically part of a clonal population of host cells. As used herein, reference to a host cell also includes a clonal population of said cells. A clonal population is a population grown from a single parent host cell. The host cell can be from any suitable organism. Suitable host cells include bacteria, fungi, or mammalian cells.

[0221] Host cells can be used to aid in the amplification of vector nucleic acids (e.g., using plasmids), or they can serve as biofactories for expressing polypeptides of the invention that form BTLA antibodies or antigen-binding fragments thereof. Suitable hosts for amplifying vector nucleic acids can be bacterial or fungal cells, such as Escherichia coli cells or Saccharomyces cerevisiae cells. Suitable hosts for expressing proteins of the invention (i.e., polypeptides that constitute human BTLA-binding antibodies or antigen-binding fragments thereof of the invention) are mammalian cells, such as HEK293T or CHO-K1 cells. In specific embodiments, the host cells are mammalian cells, such as HEK293T or CHO-K1 cells.

[0222] A variety of host-expression vector systems can be used to express the BTLA-binding molecules described herein (see, for example, U.S. Patent No. 5,807,715). For example, mammalian cells such as Chinese hamster ovary cells (CHO) are effective expression systems for CEA proteins together with vectors such as the major intermediate-early gene promoter element from human cytomegalovirus (Foecking et al., Gene, 45:101, 1986; and Cockett et al., Bio / Technology, 8:2 (1990)). Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be selected to ensure the correct modification and processing of the proteins disclosed herein. To this end, eukaryotic host cells with cellular machinery for correct processing of primary transcripts, glycosylation and phosphorylation of gene products can be used. Such mammalian host cells include, but are not limited to, CHO, HEK, VERY, BHK, Hela, COS, MDCK, 293, 3T3, W138, BT483, Hs578T, HTB2, BT20, and T47D, NSO, CRL7030, and HsS78Bst cells.

[0223] Antibody production

[0224] According to a fifth aspect of the present invention, a method for producing an antibody or antigen-binding fragment thereof according to the first aspect of the present invention is provided, comprising the steps of culturing the host cell of the fourth aspect of the present invention under conditions for producing the antibody or antigen-binding fragment thereof, and optionally isolating and / or purifying the antibody or antigen-binding fragment thereof.

[0225] According to a variation of the fifth aspect of the present invention, a method for producing an antibody or an antigen-binding fragment thereof that binds to human BTLA is provided, comprising the steps of: culturing a host cell under conditions for producing the antibody or the antigen-binding fragment thereof, the host cell comprising a nucleic acid encoding a polypeptide that forms the antibody or the antigen-binding fragment thereof that binds to human BTLA, and optionally further comprising isolating / purifying the antibody or the antigen-binding fragment thereof.

[0226] By isolated / purified is meant that the antibodies or antigen-binding fragments thereof of the invention, or polypeptides comprising these molecules, will be free or substantially free of materials with which they are naturally associated, such as other proteins or nucleic acids found in their natural environment or in the environment in which they are prepared (e.g., cell culture) when such preparation is performed by recombinant DNA techniques performed in vitro or in vivo.

[0227] According to a variation of the fifth aspect of the present invention, there is provided a method for preparing an antibody or an antigen-binding fragment thereof that specifically binds to human BTLA, the method comprising the following steps:

[0228] a) providing a host cell comprising one or more nucleic acid molecules encoding amino acid sequences of a heavy chain variable domain and a light chain variable domain that, when expressed, combine to produce a human BTLA-binding molecule;

[0229] b) culturing a host cell that expresses the encoded amino acid sequence; and c) isolating the antibody or antigen-binding fragment molecule.

[0230] The one or more nucleic acid molecules are those described above that encode polypeptides capable of forming antibodies of the present invention or antigen-binding fragments thereof that specifically bind to human BTLA.

[0231] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: i) a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence set forth in SEQ ID NO:1, CDRH2 has the amino acid sequence set forth in SEQ ID NO:2, and CDRH3 has the amino acid sequence set forth in SEQ ID NO:3; and

[0232] ii) a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO:4, CDRL2 has the amino acid sequence shown in SEQ ID NO:5, and CDRL3 has the amino acid sequence shown in SEQ ID NO:6.

[0233] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: i) a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein CDRH1 has the amino acid sequence set forth in SEQ ID NO:7, CDRH2 has the amino acid sequence set forth in SEQ ID NO:8, and CDRH3 has the amino acid sequence set forth in SEQ ID NO:9; and

[0234] ii) a light chain variable region comprising the following three CDRs: CDRL1, CDRL2 and CDRL3, wherein CDRL1 has the amino acid sequence shown in SEQ ID NO: 10, CDRL2 has the amino acid sequence shown in SEQ ID NO: 11, and CDRL3 has the amino acid sequence shown in SEQ ID NO: 12.

[0235] In certain embodiments, the antibody or antigen-binding fragment thereof comprises:

[0236] i) a heavy chain variable region comprising the amino acid sequence disclosed in SEQ ID NO: 13, or a sequence having at least 90% sequence identity thereto; and

[0237] ii) a light chain variable region comprising the amino acid sequence disclosed in SEQ ID NO: 14, or a sequence having at least 90% sequence identity thereto.

[0238] In various embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, and the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2, and CDRL3, wherein (1) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36, respectively; (2) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO:37, SEQ ID NO:38, and SEQ ID NO:39, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42, respectively. NO:42; (3) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:43, SEQ ID NO:44 and SEQ ID NO:45, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48, respectively; (4) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:43, SEQ ID NO:56 and SEQ ID NO:57, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:58, SEQ ID NO:59 and SEQ ID NO:60, respectively; (5) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:61, SEQ ID NO:62 and SEQ ID NO:63, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:64, SEQ ID NO:65 and SEQ ID NO:66, respectively. NO:66; (6) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:69, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:72, respectively;(7) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 73, SEQ ID NO: 74, and SEQ ID NO: 75, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 76, SEQ ID NO: 47, and SEQ ID NO: 78, respectively; (8) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 79, SEQ ID NO: 80, and SEQ ID NO: 81, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 82, SEQ ID NO: 83, and SEQ ID NO: 84, respectively; (9) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 85, SEQ ID NO: 86, and SEQ ID NO: 87, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 90, respectively. NO:90; (10) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:61, SEQ ID NO:92 and SEQ ID NO:93, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96, respectively; (11) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:97, SEQ ID NO:98 and SEQ ID NO:99, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:100, SEQ ID NO:101 and SEQ ID NO:102, respectively; (12) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:103, SEQ ID NO:104 and SEQ ID NO:105, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:64, SEQ ID NO: The amino acid sequences shown in SEQ ID NO:107 and SEQ ID NO:108;(13) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 109, SEQ ID NO: 110, and SEQ ID NO: 111, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 114, respectively; (14) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 85, SEQ ID NO: 110, and SEQ ID NO: 117, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID NO: 114, respectively; (15) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 124, SEQ ID NO: 125, and SEQ ID NO: 126, respectively. (16) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively; (17) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 133, SEQ ID NO: 134, and SEQ ID NO: 135, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138, respectively; (18) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO: 141, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 143, and SEQ ID NO: 144, respectively. The amino acid sequences shown in SEQ ID NO:142, SEQ ID NO:143, and SEQ ID NO:144;(19) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 145, SEQ ID NO: 146, and SEQ ID NO: 147, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 148, SEQ ID NO: 149, and SEQ ID NO: 150, respectively; (20) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 148, SEQ ID NO: 149, and SEQ ID NO: 150, respectively; (21) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 159, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138, respectively. (22) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 169, SEQ ID NO: 170, and SEQ ID NO: 171, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 172, SEQ ID NO: 173, and SEQ ID NO: 174, respectively; (23) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 61, SEQ ID NO: 62, and SEQ ID NO: 63, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 64, SEQ ID NO: 65, and SEQ ID NO: 180, respectively; (24) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 182, and SEQ ID NO: 183, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 31, SEQ ID NO: 182, and SEQ ID NO: 183, respectively. The amino acid sequences shown in SEQ ID NO:136, SEQ ID NO:137, and SEQ ID NO:186;(25) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188, and SEQ ID NO: 189, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 190, SEQ ID NO: 191, and SEQ ID NO: 192, respectively; (26) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 193, SEQ ID NO: 194, and SEQ ID NO: 195, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 196, SEQ ID NO: 197, and SEQ ID NO: 12, respectively; (27) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO: 201, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: NO:203 and SEQ ID NO:6; (28) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:205, SEQ ID NO:206 and SEQ ID NO:207, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:142, SEQ ID NO:209 and SEQ ID NO:210, respectively; (29) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:211, SEQ ID NO:212 and SEQ ID NO:213, respectively, and CDRL1, CDRL2 and CDRL3 have the amino acid sequences shown in SEQ ID NO:214, SEQ ID NO:35 and SEQ ID NO:216, respectively; (30) CDRH1, CDRH2, CDRH3 have the amino acid sequences shown in SEQ ID NO:127, SEQ ID NO:386 and SEQ ID NO:39, respectively. CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively;(31) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 205, SEQ ID NO: 206, and SEQ ID NO: 207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 209, and SEQ ID NO: 210, respectively; (32) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 388, and SEQ ID NO: 129, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively; (33) CDRH1, CDRH2, and CDRH3 have the amino acid sequences shown in SEQ ID NO: 205, SEQ ID NO: 387, and SEQ ID NO: 207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences shown in SEQ ID NO: 142, SEQ ID NO: 388, and SEQ ID NO: 36, respectively. NO:209 and SEQ ID NO:210; or (34) CDRH1, CDRH2, CDRH3 have the amino acid sequences set forth in SEQ ID NO:205, SEQ ID NO:387, and SEQ ID NO:207, respectively, and CDRL1, CDRL2, and CDRL3 have the amino acid sequences set forth in SEQ ID NO:142, SEQ ID NO:143, and SEQ ID NO:210, respectively; wherein 0 to 3 amino acid modifications may be present in any CDR / SEQ ID NO:;

[0239] Conditions for producing the antibodies or antigen-binding fragments thereof of the invention and purifying the molecules are well known in the art.

[0240] A way to solve this problem is to prepare a cell clone colony that can express antibody of the present invention or its fragment, and these cells are cultivated for a period of time in a suitable growth medium at a temperature that is conducive to cell colony expansion / growth and target protein expression.If target protein (for example antibody of the present invention) is expressed in host cells, then lysed cells (for example, using gentle detergent or ultrasonic treatment) can be used to release the content of cell (therefore target protein) into surrounding medium (which can be culture medium or another medium of reconstructing cells therein), and then the medium is carried out to purification process.If target protein (for example antibody of the present invention) is secreted into growth medium, then culture medium is carried out to purification process. Antibody purification is generally related to the method for using establishment from for example culture medium or from the culture supernatant of hybridoma cell line, separation of antibody, described method is generally related to chromatography (for example, using affinity chromatography, anion and / or cation exchange chromatography, size exclusion chromatography or other separation techniques), to separate target protein from unwanted host-derived protein and other cell contaminants (for example nucleic acid, carbohydrate etc.). Purified protein can also undergo viral inactivation step. Finally, for example, the target protein of purification can be freeze-dried or prepared in preparation for storage, transportation and subsequent use. Preferably, the protein of interest (eg, an antibody or antigen-binding fragment thereof of the invention) is substantially free of contaminating proteins initially present in the culture medium following expression or cell lysis.

[0241] In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention are at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% pure.

[0242] The protein of the present invention (the antibody or antigen-binding fragment thereof of the present invention) can be formulated into a suitable composition.

[0243] Composition

[0244] Although BTLA-binding molecules can be administered alone, in certain embodiments, administration is the administration of a pharmaceutical composition, wherein the BTLA-binding molecule is formulated with at least one pharmaceutically acceptable excipient. The excipient can be a suitable pharmaceutical carrier solute. Such carriers are well known in the art and include phosphate-buffered saline solutions, water, liposomes, various types of wetting agents, sterile solutions, and the like. Compositions containing such carriers can be formulated by well-known conventional methods. These pharmaceutical compositions can be administered to a subject at an appropriate dose. The dosage regimen will be determined by the attending physician and clinical factors.

[0245] According to a sixth aspect of the present invention, a pharmaceutical composition is provided, comprising a pharmaceutically acceptable excipient and a therapeutically effective amount of the antibody or antigen-binding fragment thereof of the first aspect of the present invention, or the antibody or antigen-binding fragment thereof produced by the fifth aspect of the present invention. In a specific embodiment, the composition comprises phosphate-buffered saline.

[0246] A "pharmaceutical composition" is a preparation that is in a form such that the biological activity of the active ingredient is effective and that does not contain additional components that are unacceptably toxic to a subject to which the formulation is administered. A pharmaceutical composition will contain one or more pharmaceutically acceptable excipients. The term excipient as used herein refers to any additive, such as a filler, solubilizer, carrier, vehicle, additive, or the like.

[0247] Pharmaceutical compositions may contain one or more pharmaceutically acceptable excipients, including, for example, water, ion exchangers, proteins, buffer substances, and salts. Preservatives and other additives may also be present. The excipient may be a solvent or a dispersion medium. Suitable formulations for the treatment methods disclosed herein are described in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980).

[0248] "Pharmaceutically acceptable" excipients are those that can be reasonably administered to a subject mammal to provide an effective dose of the active ingredient. The pharmaceutical compositions of the present invention are prepared for storage in the form of lyophilized formulations or aqueous solutions by mixing the composition with optional pharmaceutically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Acceptable excipients are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin; Protein, gelatin or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN (TM), PLURONICS (TM) or polyethylene glycol (PEG). Lyophilized HER2 antibody formulations are described in WO 97 / 04801.

[0249] Pharmaceutical compositions for in vivo administration must be sterile. This can be readily accomplished by filtration through sterile filtration membranes.

[0250] The route of administration of a BTLA-binding moiety molecule, such as an antibody, or an antigen-binding fragment thereof can be, for example, oral, parenteral, inhaled, or topical. The term parenteral as used herein includes, for example, intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, rectal, or vaginal administration.

[0251] Pharmaceutical compositions for parenteral administration include sterile aqueous solutions or non-aqueous solutions, and suspensions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, aqueous solutions, or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's (Ringers') dextrose, dextrose and sodium chloride, lactated Ringer's solution, or fixed oils. Intravenous vehicles include fluids and nutritional supplements, electrolyte supplements (such as those based on Ringer's dextrose), etc.

[0252] The injectable composition can be administered using medical devices known in the art. For example, a hypodermic needle can be used. Needle-free injection devices, such as those disclosed in U.S. Patents 6,620,135 and 5,312,335, can also be used.

[0253] Pharmaceutical compositions for oral administration can be in tablet, capsule, powder, liquid or semisolid form. Tablets can include solid carriers, such as gelatin or adjuvants. Liquid pharmaceutical compositions typically include liquid carriers, such as water, petroleum, animal or vegetable oils, mineral oils or synthetic oils. When needed, physiological saline solutions, dextrose or other sugar solutions, or glycols, such as ethylene glycol, propylene glycol or polyethylene glycol can be included.

[0254] Antibodies or their antigen-binding fragments of the present invention can be formulated into liquid, semisolid or solid forms according to the physicochemical properties and delivery routes of the molecules. The formulations can comprise a combination of excipients or excipients, such as sugars, amino acids and surfactants. Liquid formulations can have a wide range of antibody concentrations and pH values. For example, solid formulations can be produced by freeze-drying, spray drying or by the drying of supercritical fluid technology.

[0255] The pharmaceutical composition can be administered as a single dose, multiple doses, or by infusion within a defined time period. The dosage regimen can also be adjusted to provide optimal desired response (e.g., treatment or prophylactic response). In particular, parenteral formulations can be a single bolus dose, an infusion, or a loaded bolus dose, followed by one or more maintenance doses. These compositions can be administered at specific fixed or variable intervals, e.g., once a day, or "on demand."

[0256] dose

[0257] The amount of a therapeutically effective BTLA-binding molecule or pharmaceutical formulation containing such a molecule can be determined by standard clinical techniques, such as by dose-ranging clinical trials. In addition, in vitro assays can optionally be used to help determine the optimal dosage range. The exact dose to be used in the formulation will also depend on the route of administration and the severity of the disease or condition and should be determined according to the physician's judgment and the circumstances of each patient. Effective doses can be extrapolated from dose-response curves obtained from in vitro or animal model test systems. The dose of the composition to be administered can be determined by a skilled artisan in conjunction with standard dose-response studies without undue experimentation. Relevant circumstances to be considered in making these decisions include the condition or conditions to be treated, the choice of composition to be administered, the age, weight and response of the individual patient, and the severity of the patient's symptoms. For example, the actual patient weight can be used to calculate the dosage of the formulation to be administered in milliliters (mL). It may not be adjusted downward to an "ideal" weight. In such cases, the appropriate dosage can be calculated by the following formula:

[0258] Dose (ml.) = [patient weight (kg) x dose level (mg / kg) / drug concentration (mg / mL)]

[0259] The therapeutically effective dose of a pharmaceutical composition for treating a BTLA-related disease or condition as described herein will vary depending on a number of factors, including the route of administration, the target site, the patient's physiological state, the patient's weight, the patient's sex, the patient's age, whether the patient is human or an animal, other medications being administered, and whether the treatment is prophylactic or therapeutic. The therapeutically effective dose may be determined from clinical trials and may be determined by the attending physician using treatment guidelines. Typically, the patient is human, but non-human mammals may also be treated. The therapeutic dose may be titrated using conventional methods known to those skilled in the art to optimize safety and efficacy.

[0260] In various embodiments, the BTLA-binding molecule is administered at a concentration of about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg.

[0261] The pharmaceutical composition of the present invention can be administered alone or in combination with other treatments, either simultaneously or sequentially, depending on the condition to be treated. Such combinations may be combined with other immunosuppressants, for example, selected from: corticosteroids, cyclosporine, azathioprine, sulfasalazine, methotrexate, mycophenolate mofetil, tacrolimus and fingolimod, or other biologics such as infliximab, adalimumab, ustekinumab, tocilizumab and rituximab.

[0262] According to a seventh aspect of the present invention, there is provided a method for preparing a pharmaceutical composition, the method comprising formulating the antibody or antigen-binding fragment thereof according to the first aspect of the present invention, or the antibody or antigen-binding fragment thereof produced according to the fifth aspect of the present invention, into a composition comprising at least one additional component. In a particular embodiment, the at least one additional component is a pharmaceutically acceptable excipient.

[0263] Reagent test kit

[0264] In addition, products (e.g., BTLA-binding molecules or pharmaceutical compositions thereof) can be packaged and sold in the form of a kit. Such articles of manufacture can have a label or package insert indicating instructions for the product and its appropriate use for treating a subject suffering from or susceptible to a disease or condition.

[0265] Thus, according to one aspect of the present invention, there is provided a kit comprising an antibody or antigen-binding fragment thereof according to the first aspect of the invention or a pharmaceutical composition according to the sixth aspect of the invention. Suitably, such a kit includes a package insert containing instructions for use.

[0266] Therapeutic / medical uses

[0267] The antibodies or antigen-binding fragments thereof of the present invention or pharmaceutical compositions comprising the same can be used for therapy, typically as a medicament.

[0268] In certain embodiments, the antibodies or antigen-binding fragments thereof of the present invention, or pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof, can be used to treat or prevent any disease or condition in a subject in need thereof.

[0269] BTLA is involved in downregulating immune responses, and there are many diseases or conditions that can be treated by inhibiting host T cells and / or B cells (e.g., see Crawford & Wherry. Editorial: Therapeutic potential of targeting BTLA. J Leukocyte Biol. 86: 5-8, 2009). Diseases or conditions that can benefit from treatment with anti-BTLA agonists are referred to herein as "BTLA-associated diseases." BTLA-associated diseases include inflammatory or autoimmune diseases, as well as conditions with excessive immune cell proliferation.

[0270] Specific BTLA-associated diseases that can be treated with the BTLA-binding molecules of the invention include: Addison's disease, allergies, alopecia areata, amyotrophic lateral sclerosis, ankylosing spondylitis, antiphospholipid syndrome, asthma (including allergic asthma), autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune pancreatitis, autoimmune polyendocrine syndrome, Behcet's disease, bullous pemphigoid, cerebral malaria, chronic inflammatory demyelinating polyneuropathy, celiac disease, Crohn's disease, Cushing's syndrome, dermatomyositis, type 1 diabetes mellitus, eosinophilic granulomatosis with polyangiitis, graft-versus-host disease, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease,

[0015] Examples of the drugs listed include: leukemia, lymphoma, lymphoproliferative disorders, multiple sclerosis, myasthenia gravis, myeloma, neuromyelitis optica, pemphigus, polymyositis, primary biliary cholangitis, primary sclerosing cholangitis, psoriasis, psoriatic arthritis, rheumatoid arthritis, sarcoidosis, Sjögren's syndrome, systemic lupus erythematosus, Takayasu's arteritis, temporal arteritis, transplant rejection, transverse myelitis, ulcerative colitis, uveitis, vasculitis, vitiligo, and Vogt-Koyanagi-Harada disease.

[0271] According to an eighth aspect of the present invention, there is provided an antibody or antigen-binding fragment thereof according to the first aspect of the present invention or a pharmaceutical composition according to the sixth aspect of the present invention for use in therapy.

[0272] In certain embodiments, the treatment is the treatment or prevention of a BTLA-associated disease.

[0273] In certain embodiments, a BTLA-associated disease is a disease caused by decreased expression and / or activity of BTLA in a subject. In particular, any disease or condition characterized by the presence or activity of T or B cells can be treated with the BTLA agonist antibodies or antigen-binding fragments of the invention.

[0274] In one embodiment, the BTLA-associated disease is an inflammatory disease (eg, rheumatoid arthritis), an autoimmune disease or disorder (eg, graft-versus-host disease), or a proliferative disease or disorder (eg, cancer).

[0275] In certain embodiments, the treatment is the treatment or prevention of inflammatory or autoimmune diseases and disorders of excessive immune cell proliferation.

[0276] According to variations of the eighth aspect of the present invention, a method of treating a patient in need thereof is provided, comprising administering to the patient an antibody or antigen-binding fragment thereof according to the first aspect of the present invention, or a pharmaceutical composition according to the sixth aspect of the present invention. In certain embodiments, the patient in need of treatment or to be treated suffers from (or is suffering from) a BTLA-associated disease. In certain embodiments, the patient in need of treatment or to be treated suffers from (or is suffering from) an inflammatory disease, an autoimmune disease, or a condition characterized by excessive immune cell proliferation.

[0277] In a specific embodiment, the antibody or antigen-binding fragment thereof according to the first aspect of the invention or the pharmaceutical composition according to the sixth aspect of the invention is administered to a patient in need thereof in a pharmaceutically acceptable amount.

[0278] In variations of this aspect, provided are antibodies or antigen-binding fragments thereof according to the first aspect of the invention or pharmaceutical compositions according to the sixth aspect of the invention for use in a method of treating a patient in need thereof. In certain embodiments, the method is used to treat or prevent a BTLA-associated disease. In certain embodiments, the method is used to treat or prevent an inflammatory or autoimmune disease, as well as conditions of excessive immune cell proliferation.

[0279] In a further variation of this aspect, there is provided use of an antibody or antigen-binding fragment thereof according to the first aspect of the invention or a pharmaceutical composition according to the sixth aspect of the invention in the preparation of a medicament for treating a patient in need thereof.

[0280] In one embodiment, the treatment is for treating a BTLA-associated disease. Suitably, the BTLA-associated disease is an inflammatory disease (such as asthma), an autoimmune disease or disorder (such as rheumatoid arthritis) or an immunoproliferative disease or disorder (such as lymphoma).

[0281] In certain embodiments, the antibodies or antigen-binding fragments thereof of the present invention or pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof are used to inhibit T cells and / or B cells.

[0282] In certain embodiments, the antibodies or antigen-binding fragments thereof of the present invention, or pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof, are used to treat or prevent a disease or condition in a subject in need thereof, the disease or condition being selected from the group consisting of Addison's disease, allergy, alopecia areata, amyotrophic lateral sclerosis, ankylosing spondylitis, antiphospholipid syndrome, asthma (including allergic asthma), autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune pancreatitis, autoimmune polyendocrine syndrome, Behcet's disease, bullous pemphigoid, cerebral malaria, chronic inflammatory demyelinating polyneuropathy, celiac disease, Crohn's disease, Cushing's syndrome, dermatomyositis, type 1 diabetes, eosinophilic granulomatosis with polyangiitis, graft-versus-host disease (GVHD), leukemia, myeloma, leukemia, leukemia, schisis, leukemia, necrosis factor-α ... GVHD), Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis, hidradenitis suppurativa, inflammatory fibrosis (e.g., scleroderma, pulmonary fibrosis, and cirrhosis), juvenile arthritis, Kawasaki disease, leukemia, lymphoma, lymphoproliferative disorders, multiple sclerosis (MS), myasthenia gravis, myeloma, neuromyelitis optica, pemphigus, polymyositis, primary biliary cholangitis, primary sclerosing cholangitis, psoriasis, psoriatic arthritis, rheumatoid arthritis, sarcoidosis, Sjögren's syndrome, systemic lupus erythematosus, Takayasu's arteritis, temporal arteritis, transplant rejection, transverse myelitis, ulcerative colitis, uveitis, vasculitis, vitiligo, and Vogt-Koyanagi-Harada disease.

[0283] In certain embodiments, the antibodies or antigen-binding fragments thereof of the present invention, or pharmaceutical compositions comprising the same, are used to treat or prevent a disease or condition in a subject in need thereof, the disease or condition being selected from the group consisting of: GVHD, colitis, rheumatoid arthritis, psoriasis, and MS. In one embodiment, the immunoproliferative disease is cancer. Suitably, the cancer is leukemia or lymphoma.

[0284] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof is used to prevent or treat transplant rejection.

[0285] In another embodiment, the present invention relates to the prevention or treatment of graft-versus-host disease.

[0286] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof is used to treat rheumatoid arthritis.

[0287] In other embodiments, the antibodies or antigen-binding fragments thereof of the present invention, or pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof, are used to treat diabetes, such as type 1 diabetes.

[0288] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof is used to treat psoriasis.

[0289] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof is used to treat multiple sclerosis.

[0290] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof is used to treat colitis.

[0291] The term "effective amount" or "therapeutically effective amount" refers to a dose or amount of a drug sufficient to improve symptoms in a patient or to achieve a desired biological outcome (e.g., for cancer, increased tumor cell death, decreased tumor size, prolonged progression-free survival or overall survival, etc.). As disclosed elsewhere herein, an effective amount will generally be assessed through extensive human clinical studies.

[0292] Throughout the specification and claims of this application, the words "comprise" and "contain" and their variations mean "including but not limited to", and they are not intended to (and do not) exclude other parts, additives, components, integers or steps. Throughout the specification and claims of this application, unless the context requires otherwise, the singular encompasses the plural. In particular, where the indefinite article is used, the description should be understood as contemplating plurality as well as singularity unless the context requires otherwise.

[0293] Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all steps of any method or process disclosed, may be combined in any combination, except where at least some of such features and / or steps are mutually exclusive. The invention is not limited to the details of any foregoing embodiments. The invention extends to any novel feature or any novel combination of features disclosed in this specification (including any accompanying claims, abstract and drawings), or any novel step or any novel combination of steps of any method or process so disclosed.

[0294] The reader's attention is directed to all papers and documents filed concurrently with or prior to the specification in connection with this application, which are open to public inspection with respect to this specification, and the contents of all such papers and documents are incorporated herein by reference.

[0295] The invention will now be further described with reference to the following non-limiting examples and figures. BRIEF DESCRIPTION OF THE DRAWINGS

[0296] Figure 1

[0297] Antibody binding to soluble and cell-expressed forms of human and cynomolgus monkey BTLA. (a) Surface plasmon resonance (SPR) binding curves of soluble monomeric human BTLA ectodomain injected at a higher concentration than immobilized anti-BTLA antibody; the graph shows the SPR signal after reference and blank subtraction. (b) Association and dissociation rates for binding to human or cynomolgus monkey BTLA, as calculated by curve fitting using BiaEvaluation software. (c) Binding of antibody 2.8.6 to Jurkat cell lines expressing human BTLA or cynomolgus monkey BTLA, compared to an isotype control antibody. (d) EC50 for antibody binding to transfected cell lines, as calculated by nonlinear curve fitting using GraphPad Prism software.

[0298] Figure 2

[0299] (a) Blockade of ligand binding by anti-BTLA antibodies assessed by SPR. The human BTLA extracellular domain was immobilized on a sensor chip. Human HVEM was injected to confirm binding and then allowed to completely dissociate. A saturating concentration of anti-BTLA antibody was then injected, immediately followed by a second injection of HVEM. (b) Equilibrium binding of HVEM after antibody injection is expressed as a percentage of HVEM binding before antibody injection. Saturation of BTLA with clone 11.5.1 blocked subsequent ligand binding, while saturation with clone 2.8.6 did not block.

[0300] Figure 3

[0301] Epitope mapping of anti-BTLA antibodies. (a) HEK293T cells transfected with a BTLA construct in a bicistronic vector that also expresses GFP were stained with a Pacific Blue-conjugated anti-BTLA antibody. Clone 11.5.1 binds to cells transfected with the wild-type receptor (left), but not to cells transfected with BTLA with the Y39R mutation (right). (b) For clones 2.8.6 and 11.5.1, binding to each BTLA mutant construct is expressed as a percentage relative to binding to the wild-type receptor. (c) Mutations Y39R and K41E, which selectively abolished binding of clone 11.5.1, were mapped to the crystal structure of human BTLA (black residues). Residues critical for ligand HVEM binding are highlighted in gray.

[0302] Figure 4

[0303] (a) Crystal structure of the human BTLA ectodomain in complex with the Fab' fragment of clone 2.8.6. Residues on BTLA buried at the interface are highlighted in black. (b) The epitope of antibody 2.8.6 (black residues) is shown relative to the HVEM binding site (grey residues).

[0304] Figure 5

[0305] (a) Strategy for generating a chimeric BTLA gene in humanized BTLA mice. A segment of human genomic DNA from the start of exon 2 to the end of exon 3 was inserted into the mouse locus, replacing the mouse sequence from the start of exon 2 to the end of exon 4. The sequence at the exon-intron junction at the start of mouse exon 2 and the end of mouse exon 4 was left intact to ensure correct splicing.

[0306] Figure 6

[0307] (a) Scheme of the T cell transfer assay for in vivo evaluation of anti-BTLA antibodies. A mixture of humanized and wild-type OVA-specific CD4 T cells was injected into recipient mice. The next day, mice were immunized with ovalbumin in alum to activate the transferred cells and 24 hours later were given anti-human BTLA antibodies or an isotype control. Eight days after the initial cell transfer, the ratio of humanized cells to wild-type cells in the transferred population in the spleen was assessed by flow cytometry. (b) Both clones 11.5.1 and 2.8.6 (to a lesser extent) reduced the expansion of humanized cells relative to wild-type. The graph shows the pooled data from two (for 11.5.1) or three (for 2.8.6) replicates.

[0308] Figure 7

[0309] Effect of anti-BTLA clone 2.8.6 on CD4 T cell proliferation in an in vitro mixed lymphocyte reaction. T cells from humanized C57BL / 6 mice were stained with CellTrace Violet and added to mitomycin C-treated Balb / c stimulator cells in the presence of anti-BTLA antibody or an isotype control. After 96 hours, proliferation of humanized CD4 cells was assessed and normalized to that in the absence of antibody. Clone 2.8.6 inhibited proliferation of humanized cells with an IC50 of 0.029 nM, with a maximal effect of 42% proliferation inhibition.

[0310] Figure 8

[0311] (a) Effect of clone 2.8.6 in a T cell colitis model. RAG knockout recipient mice were injected with CD45RBhiCD25-CD4+ T cells from humanized BTLA mice and treated with 200 μg of 2.8.6 or isotype control antibody on days 7, 21, and 35. Isotype control-treated mice gradually lost weight starting from week 3, while 2.8.6-treated mice were unaffected. (b) Eight weeks after cell transfer, the colons were processed to extract lamina propria lymphocytes, and the total number of inflammatory cells extracted per colon was counted. Isotype control-treated mice had significantly more infiltrating immune cells than 2.8.6-treated mice. (c) The colon weight to length ratio was calculated as a marker of inflammation and thickening. 2.8.6 treatment prevented the increase in weight to length ratio observed in isotype control-treated mice.

[0312] Figure 9

[0313] (a) Effect of BTLA antibodies in the parental-F1 model of GVHD. C57BL / 6 splenocytes and bone marrow cells from humanized BTLA mice were injected into CB6F1 recipient mice, which were then treated with anti-BTLA antibodies or isotype control. Untreated mice developed clinical GVHD with progressive weight loss, dermatitis, and diarrhea and were withdrawn upon reaching a pre-specified humane endpoint. Mice treated with 2.8.6 and 11.5.1 antibodies were relatively unaffected, with survival rates comparable to control mice reconstituted with syngeneic cells. (b) Five weeks after cell transfer, mice were withdrawn, and the ratio of colon weight to length was calculated as a marker of intestinal inflammation. Treatment with 2.8.6 and 11.5.1 prevented the colon thickening observed in untreated mice.

[0314] Figure 10

[0315] (a) Effect of clone 11.5.1 with the D265A mutation in an in vivo T cell transfer assay. This mutant antibody, which does not bind to Fc receptors, no longer inhibits the proliferation of humanized BTLA cells, but rather enhances proliferation due to receptor blockade. (b) The D265A mutation in clone 11.5.1 no longer inhibits T cell proliferation in a mixed lymphocyte reaction.

[0316] Figure 11

[0317] Anti-BTLA antibodies do not fix complement. Splenocytes from humanized BTLA mice were incubated with 10% rabbit complement for 1 hour at 37°C in the presence of 20 μg / ml BTLA antibody, an isotype control, or a positive control (a CD20-depleting antibody). The anti-CD20 antibody depleted most B cells, confirming the activity of rabbit complement, but the BTLA antibody did not deplete B or T cells, even though both populations stained positive for BTLA.

[0318] Figure 12

[0319] Anti-BTLA antibodies do not induce antibody-dependent cell-mediated cytotoxicity. Splenocytes from humanized BTLA mice were incubated at 37°C for 24 hours in the presence of 20 μg / ml BTLA antibody, isotype control, or a positive control (CD20-depleting antibody). Anti-CD20 antibodies depleted most B cells by inducing ADCC on effector cells in the mixture, but BTLA antibodies did not deplete B or T cells, even though both populations stained positive for BTLA.

[0320] Figure 13

[0321] Anti-BTLA antibodies do not deplete B or T cells in vivo. Humanized BTLA mice were injected with 200 μg of 2.8.6 antibody. At 24 hours, spleens and bone marrow were harvested, and cell populations were assessed by flow cytometry. 2.8.6 did not deplete B or T cells in the spleen or affect the frequencies of different B cell precursor populations in the bone marrow.

[0322] Figure 14

[0323] B cells or CD4 T cells from humanized mice after 6 days of incubation with antibodies 2.8.6 or 11.5.1 compared to BTLA expression levels on cells from mice injected with an isotype control antibody. + BTLA expression levels on T cells.

[0324] Example

[0325] In the following examples, antibodies 11.5.1 and 2.8.6 are shown to bind human BTLA with high affinity. Using transgenic mice expressing the human receptor, it was demonstrated that upon binding to BTLA, these antibodies inhibited T cell responses in vitro and in vivo and were able to ameliorate disease in mouse models of inflammatory bowel disease and graft-versus-host disease. Although these agonist effects were dependent on Fc receptor binding, these antibodies did not lead to the depletion of BTLA-expressing cells through cytotoxicity, nor did they induce receptor downregulation.

[0326] Example 1. Generation and sequencing of anti-BTLA antibodies

[0327] The antibody that recognizes the human immune cell receptor BTLA was produced by BioGenes GmbH using the extracellular region of human BTLA (BTLA K31-R151) were immunized with mice. Splenocytes from these immunized mice were fused with Sp2 / 0-Ag14 myeloma cells, and the resulting hybridomas were selected for reactivity with human BTLA by ELISA of the supernatants combined with dilution cloning. Antibodies were isotyped from hybridoma supernatants using the Rapid Mouse Isotyping Kit (RayBiotech). The antibodies produced by clones 2.8.6 and 11.5.1 were both IgG1k.

[0328] For sequencing of immunoglobulin variable domains, RNA was extracted from hybridomas using TRIzol reagent (ThermoFisher) according to the manufacturer's instructions. RNA was reverse transcribed using primers specific for the first heavy chain constant domain or the light chain constant domain and Super Script II reverse transcriptase (Invitrogen) according to the manufacturer's instructions to generate cDNA.

[0329] Then, as previously described (Tiller et al., J Immunol Methods. 350: 183-193, 2009), PCR was performed using primers targeting conserved regions of the immunoglobulin locus, and the PCR products were sequenced. In some cases, a large amount of non-functional kappa light chain cDNA from fusion myeloma cell lines complicated the identification of functional light chains. To address this issue, a previously described technique was used to add an excess of primers specific for the non-functional chain CDR3 to force truncation of the abnormal chain product (Yuan et al., J Immunol Methods. 294: 39553-61, 2005).

[0330] The variable domain sequences were evaluated using the NCBI IgBlast tool to determine the locations of the CDRs.

[0331] Example 2. Binding to soluble human and cynomolgus monkey BTLA

[0332] The binding affinity and kinetics of the BTLA agonist antibodies of the present invention (2.8.6 and 11.5.1) to human or cynomolgus monkey BTLA were determined by surface plasmon resonance using a Biacore T200 (GE Healthcare). A Series S CM5 Sensor Chip (GE Healthcare) was coated with polyclonal anti-mouse IgG using a mouse antibody capture kit (GE Healthcare). The anti-BTLA antibody was then captured onto the biosensor surface, and a negative control antibody (clone Mopc21; Biolegend) was captured in the reference channel. Various concentrations of monomeric soluble human BTLA extracellular domain (BTLA) were then added to the biosensor surface in a single cycle kinetic analysis at 37°C.K31-R151 ) (from SEQ ID NO: 23) or soluble cynomolgus monkey BTLA ectodomain (BTLA K31-R151 ) (from SEQ ID NO: 24) was injected onto the immobilized antibody in buffer 10 mM Hepes, 150 mM NaCl, 0.005% v / v surfactant P20, pH 7.4 (HBS-P) ( Figure 1 a). After subtracting the reference and blank, the association and dissociation rates were fitted using BiaEvaluation software (GE Healthcare), and the dissociation constant ( Figure 1 b). Clone 2.8.6 bound to human BTLA with a KD of 0.65 nM and to cynomolgus monkey BTLA with a KD of 7.89 nM. Clone 11.5.1 bound to human BTLA with a KD of 0.75 nM and to cynomolgus monkey BTLA with a KD of 0.99 nM. In a separate experiment targeting only human BTLA, clone 2.8.6 bound to human BTLA with a KD of 0.37 nM, while clone 11.5.1 bound to human BTLA with a KD of 0.53 nM.

[0333] Example 3. Binding to BTLA on cells

[0334] The ability of the BTLA agonist antibodies of the present invention (2.8.6 and 11.5.1) to bind to human or cynomolgus monkey BTLA expressed on the cell surface was evaluated by flow cytometry. Full-length human or cynomolgus monkey BTLA was expressed in Jurkat T cell lines using a lentiviral transfection system. 1×10 5Cells were seeded in 96-well U-bottom plates. Starting from a concentration of 90 μg / ml, the binding of BTLA antibodies relative to the mIgG1 isotype control (clone MOPC-21, Biolegend #400165) was evaluated at 12 concentrations by 1 / 3 serial dilution in FACS buffer (PBS, 2% FCS, 0.05% sodium azide). Nonspecific antibody binding was blocked by the addition of Fc blocker (Biolegend #101319). The antibodies were incubated with cells on ice for 30 minutes, and then the cells were washed twice with FACS buffer and stained with AF647-conjugated anti-mIgG1 secondary antibody (Biolegend #406618). The secondary antibody was incubated on ice for 30 minutes, and then the cells were washed and resuspended in FACS buffer for analysis on a flow cytometer. The geometric mean fluorescence intensity of the secondary antibody was plotted for each concentration, and the EC50 of receptor binding was calculated by nonlinear curve fitting using GraphPad Prism software. Clone 11.5.1 bound to cells expressing human BTLA with an EC50 of 0.016 nM and to cells expressing cynomolgus monkey BTLA with an EC50 of 0.0057 nM. Clone 2.8.6 bound to cells expressing human BTLA with an EC50 of 0.085 nM and to cells expressing cynomolgus monkey BTLA with an EC50 of 0.16 nM. Figure 1 cd).

[0335] Example 4. Competition with the natural ligand HVEM for binding to BTLA

[0336] The ability of the BTLA agonist antibodies of the present invention (2.8.6 and 11.5.1) to block the binding of the natural ligand to BTLA was assessed by surface plasmon resonance using a Biacor eT200 (GE Healthcare). 31K-151R ) was covalently coupled to a CM5 sensor chip. The human HVEM extracellular domain fused to mouse IgG1 Fc was then injected onto immobilized hBTLA in HBS-P buffer at 37°C and allowed to completely dissociate. A saturating amount of anti-BTLA antibody (2.8.6 or 11.5.1) was then injected, followed immediately by a second injection of human HVEM-mFc ( Figure 2 a). Equilibrium HVEM binding (in resonance units) after BTLA saturation with antibody is expressed as a percentage of binding before antibody injection ( Figure 2 b) If HVEM binding after saturation with antibody is greater than 90% of the binding before antibody injection, the antibody is considered non-blocking.

[0337] Example 5. Binding epitope of antibody 11.5.1 on human BTLA

[0338] The functional epitope of antibody 11.5.1 on human BTLA was determined by flow cytometry assessment of binding to a panel of single-residue mutants of the receptor expressed on the cell surface. A construct encoding the human extracellular region of BTLA and the transmembrane and intracellular regions of murine CD28 was cloned into the bicistronic mammalian expression vector pGFP2-n2 (BioSignal Packard Ltd), which also encodes GFP. Mutant constructs differing by one amino acid were generated using the "drastic" mutagenesis method (Davis et al., Proc Natl Acad Sci USA. 95, 5490-4 (1998)). Plasmids (2 μg / well) were transfected into HEK-293T cells in 6-well plates using Genejuice transfection reagent (Novagen; 6 μl / well). Mock and non-transfected controls were included in each experiment. At 48 hours, cells were harvested and stained with 10 μg / ml fluorescent dye-conjugated anti-BTLA antibody in PBS, 0.05% azide, 2% FCS (FACS buffer) for 1 hour at 4°C, next to the live / dead marker. Cells were washed, pelleted, and resuspended in 200 μl FACS buffer before analysis on a BD FACSCanto flow cytometer. GFP-positive (transfected) live cells were gated and analyzed for binding of anti-BTLA antibody (an example of binding analysis for clone 11.5.1 is given in Figure 3 For each mutant, the geometric mean of anti-BTLA antibody binding to transfected cells was expressed as a percentage of binding to the wild-type receptor ( Figure 3 b) A panel of anti-BTLA antibodies was evaluated, and any mutation that abolished binding of all antibodies was excluded from the analysis, assuming that such mutations would result in drastic changes in protein folding or expression rather than in the antibody epitope. Mutations Y39R and K41E completely abolished binding of antibody 11.5.1, while not affecting binding of 2.8.6. These mutations were not found in Figure 3 c is mapped to the human BTLA crystal structure (Compaan et al., J Biol Chem. 280:39553-61, 2005) (black residues), indicating the binding epitope of 11.5.1. The residues required for HVEM binding (Gln37, Arg42, Pro59, His127; from patent publication number WO2017004213) are also mapped to the structure in gray, indicating that 11.5.1 binds to an epitope very close to the HVEM binding site.

[0339] Example 6. Crystal structure of the Fab′ fragment of 2.8.6 in complex with human BTLA

[0340] The structural epitope of antibody 2.8.6 on human BTLA was determined by analyzing the crystal structure of the antibody Fab in complex with the extracellular domain of human BTLA. The heavy and light chain variable domains of antibody 2.8.6 were cloned into pOPINVH and pOPINVL expression vectors (Addgene), which encode the first constant domain of mouse IgG1 heavy chain (with 6x histidine tag) and the constant domain of mouse Ig kappa chain, respectively. These vectors were transiently co-transfected into HEK293T cells to produce Fab' fragments against BTLA2.8.6, which were purified by Ni-NTA purification. S33-D135 ) was cloned into the pGMT7 vector and expressed in BL21(DE3)pLysS Escherichia coli cells (Novagen) to produce inclusion bodies. Inclusion bodies were isolated from the cell pellet by sonication and repeatedly washed with a wash buffer containing 0.5% Triton X-100. Purified BTLA inclusion bodies were solubilized in a denaturant solution containing 6 M guanidine hydrochloride. The solubilized protein solution was slowly diluted in refolding buffer [0.1 M Tris-HCl (pH 8.0), 0.6 M L-arginine, 2 mM ethylenediaminetetraacetic acid, 3.73 mM cystamine, and 6.73 mM cysteamine] to a final protein concentration of 1–2 μM and then stirred at 4°C for 48 hours. The BTLA refolding mixture was then concentrated using a VIVA FLOW50 system (Sartorius). BTLA was purified by gel filtration on a Superdex 75 column (GE Healthcare).

[0341] Purified BTLA and Fab' were mixed and purified as a complex by size exclusion chromatography. Crystals suitable for data collection were obtained by hanging drop vapor diffusion at 293°K in 0.2 M calcium acetate, 0.1 M imidazole, pH 8.0, 10% (w / v) PEG 8000. The final dataset was collected at the Photon Factory, and the structure was determined by molecular replacement using the structures of BTLA (PDB ID: 2AW2 chain A) and anti-PD1-Fab (PDB ID: 5GGS chain C, D) as search probes.

[0342] The residues on BTLA at the interface with antibody 2.8.6 are A50, G51, D52, P53, E83, D84, R85, Q86, E103, P104, V105, L106, P107, N108, and D135.

[0343] Example 7. Development of humanized BTLA mice

[0344] To provide a platform for evaluating anti-human BTLA antibodies in a mouse model, a knock-in strain of C57Bl / 6 mice was developed that expresses a chimeric form of BTLA with a human extracellular region and a mouse transmembrane and signaling region. A segment of human genomic DNA from the start of exon 2 to the end of exon 3 was inserted into the mouse locus, replacing the mouse sequence from the start of exon 2 to the end of exon 4. The sequence at the exon-intron junction at the start of mouse exon 2 and the end of mouse exon 4 was left intact to ensure correct splicing ( Figure 5 ).

[0345] Example 8. In vivo inhibition of antigen-specific T cell proliferation

[0346] A sensitive T cell transfer assay was used to evaluate the ability of the BTLA agonist antibodies of the invention (2.8.6 and 11.5.1) to inhibit the proliferation of antigen-specific T cells in vivo ( Figure 6 a). In this experiment, 5x10 5 T cells containing purified ovalbumin (OVA)-specific OTII (TCR transgenic) CD4 + A mixture of T cells from mice expressing homozygous human BTLA (hBTLA) and from OT-II mice expressing the wild-type murine BTLA receptor (The Jackson Laboratory) were used. The CD45.2 (relative to CD45.1) allotypic marker was used to distinguish the transferred cells from the host cells. The wild-type donor cells also expressed green fluorescent protein under the control of the human ubiquitin C promoter to distinguish them from the humanized donor cells by flow cytometry. The day after the T cell transfer, the recipient mice were immunized with 100 μg of ovalbumin (Sigma-Aldrich) in 100 μl of PBS mixed with 100 μl of injection alum (ThermoFisher) to induce T cell expansion. The next day, 200 μg of antibody were administered intraperitoneally to the mice. Eight days after the initial T cell transfer, the proportion of T cells expressing humanized BTLA and wild-type OVA-specific T cells in the spleen was determined by flow cytometry. In this way, the expansion or reduction of humanized cells that bind anti-human BTLA antibodies can be tracked relative to wild-type controls that do not bind anti-human BTLA antibodies. Antibodies 2.8.6 and 11.5.1 both resulted in a reduction in the expansion of humanized BTLA cells relative to wild-type controls, suggesting that they induce signaling through inhibitory BTLA receptors, leading to a reduction in T cell proliferation ( Figure 6 b).

[0347] Example 9. Inhibition of T cell proliferation in mixed lymphocyte reaction

[0348] The ability of the BTLA agonist antibodies of the present invention (2.8.6 and 11.5.1) to inhibit the proliferation of primary T cells from humanized mice in vitro was evaluated using a mixed lymphocyte reaction (MLR). Splenocytes from Balb / c mice were treated with mitomycin C for 30 minutes at 37°C, then washed and used as stimulator cells. T cells were purified from the spleens of humanized BTLA mice by negative selection using magnetic activated cell sorting (Mojosort Mouse CD3 T Cell Isolation Kit, Biolegend #480023) and stained with the CellTrace Violet Cell Proliferation Kit (ThermoFisher) to be used as responder cells. 4x10 cells per well 5 stimulator cells and 2x10 5 Responder cells were mixed with various concentrations of anti-BTLA or isotype control antibodies (clone MOPC-21, Biolegend #400165) in 96-well U-bottom plates. Serial 1 / 3 dilutions of the antibody were evaluated starting at a concentration of 1 μg / ml for a total of 10 concentrations. Polyclonal anti-mHVEM antibody (R&D systems #AF2516) was also added to all wells at 1 μg / ml to block any baseline signaling through the BTLA pathway and enhance the effect of the agonist antibody. After 96 hours, the dilution of CellTrace Violet in the responder cells was assessed by flow cytometry as a marker of proliferation. Proliferation in the presence of anti-BTLA antibody or isotype control was compared to proliferation in the absence of antibody. + and CD8 + The populations were gated and analyzed separately. Antibodies 2.8.6 and 11.5.1 both reduced the proliferation of T cells expressing human BTLA, indicating that they induce inhibitory signaling through the human BTLA receptor. Clone 2.8.6 inhibited CD4 T cells with an IC50 of 0.029 nM and had a maximal effect of 42% proliferation inhibition ( Figure 7 Clone 11.5.1 inhibited CD4 T cells with an IC50 of 0.016 nM, with a maximal effect of 33% proliferation inhibition.

[0349] Example 10. Inhibition of NFkB signaling in Jurkat T cell lines transfected with human BTLA or cynomolgus monkey BTLA

[0350] The ability of the BTLA agonist antibodies of the present invention (2.8.6 and 11.5.1) to inhibit NFkB signaling was evaluated using a BTLA-transfected reporter T cell line. A Jurkat T cell line stably transfected with an expression cassette comprising an NF-κB-responsive transcriptional element upstream of a minimal CMV promoter (mCMV)-GFP cassette (Source BioSciences #TR850A-1) was used as a reporter cell line for NFkB signaling. Full-length human or cynomolgus monkey BTLA was expressed in this reporter cell line using a lentiviral transfection system. These cells were mixed with a stimulator cell line consisting of bw5147 cells expressing an anti-CD3 ScFv construct on their surface, as described in Leitner et al. J Immunol Methods. 2010 Oct 31; 362(1-2): 131-41. This stimulator cell line was also transfected with mouse FcγRIIB to provide Fc receptors for presentation of the agonist BTLA antibody. 5×10 cells / well were added in the presence of different concentrations of BTLA antibody or isotype control (clone MOPC-21, Biolegend #400165). 4 reporter cells and 5x10 4 Stimulator cells were mixed in a 96-well U-bottom plate. After incubation at 37°C for 24 hours, the cells were pelleted and stained with a viability dye (Zombie Aqua, Biolegend #423101) and a mouse CD45 antibody (Pe-Cy7 conjugated clone 104, Biolegend #109830) for flow cytometry to separate stimulator cells (mouse) from responder (human) cells. The geometric mean of GFP expression was assessed for each antibody concentration and normalized to GFP expression in the absence of antibody. Clone 2.8.6 inhibited human BTLA-transfected cells with an IC50 of 0.06 nM and inhibited cynomolgus monkey BTLA-transfected cells with an IC50 of 0.22 nM. Clone 11.5.1 inhibited human BTLA-transfected cells with an IC50 of 0.033 nM and inhibited cynomolgus monkey BTLA-transfected cells with an IC50 of 0.14 nM.

[0351] Example 11. Treatment of T cell-driven mouse colitis model with antibody 2.8.6

[0352] The ability of the BTLA agonist antibody 2.8.6 to improve a T cell-driven colitis model was evaluated using humanized mice. This T cell transfer model has been previously described as a mouse model of inflammatory bowel disease (Ostanin et al., Am J Physiol Gastrointest Liver Physiol. 296: G135-46, 2009). CD45RB was isolated from the spleen and lymph nodes of humanized BTLA mice. hiCD25-CD4+ T cells were expressed at 5x10 per mouse. 5 A dose of 10 cells was intraperitoneally injected into Rag1 KO recipients (Rag1 tm1Mom ; The Jackson Laboratory). The transferred T cells caused inflammatory colitis, which occurred after about 3 weeks and resulted in diarrhea and weight loss. Animals in the Rag1 KO cage that did not receive the transferred T cells were used as non-diseased controls. On days 7, 21, and 35 after the T cell transfer, 200 μg of 2.8.6 or an isotype control antibody were injected intraperitoneally into the recipient mice. All mice were weighed regularly, and at week 8, the colon was weighed and measured, and inflammatory infiltration was assessed by histology, cell counts, and flow cytometric analysis of extracted lamina propria leukocytes. Antibody 2.8.6 prevented weight loss ( Figure 8 a) and significantly reduced inflammatory infiltration of the colon ( Figure 8 b). Colonic inflammation in diseased mice resulted in an increased colon weight:length ratio that was not seen in 2.8.6 treated mice ( Figure 8 c).

[0353] Example 12. Treatment of Graft-Versus-Host Disease (GVHD) Mouse Model

[0354] The effects of anti-BTLA agonist antibodies were evaluated in a non-lethal parental-F1 model of GVHD. Humanized BTLA donor mice (C57BL / 6 background; H2 b ) were collected from bone marrow cells (BMC) and spleen cells. 2x10 7 BMC and 10 7 splenocytes were injected intravenously into CB6F1(H2 b / d ) recipients. Irradiated CB6F1 mice reconstituted with syngeneic BMCs and splenocytes served as non-diseased controls. On the day of immune cell transfer, mice were injected intraperitoneally with 200 μg of anti-BTLA antibody or isotype control. Mice were weighed regularly and GVHD was monitored by calculating relative weight loss and clinical observations. Mice were withdrawn 5 weeks after immune cell transfer or when they reached a humane endpoint (including weight loss >20% relative to initial body weight in the first 14 days, or weight loss >15% at any other time). The colon was weighed and measured at the time of death, and the colon weight:length ratio was calculated as a marker of colonic inflammation, a prominent clinical feature of GVHD. Both antibodies 2.8.6 and 11.5.1 significantly reduced weight loss, resulting in increased survival ( Figure 9 a), and prevent colon inflammation ( Figure 9 b).

[0355] Example 13. Agonist activity of antibody 11.5.1 is dependent on Fc receptor binding

[0356] Antibody 11.5.1 was recombinantly expressed as a mIgG1k containing the D265A mutation, which has been previously described to significantly reduce Fc receptor binding (Clynes et al., Nat Med. 6:443-446, 2000). This mutant antibody was evaluated in the T cell transfer assay described in Example 8. The parental 11.5.1 antibody inhibited the proliferation of humanized T cells because its net effect was agonism of the BTLA receptor. However, the FcR-null D265A mutation resulted in enhanced proliferation of humanized T cells, suggesting that the FcR-null mutation abolished the agonistic effect of the antibody, leaving only a receptor-blocking effect ( Figure 10 a).

[0357] The D265A mutated 11.5.1 antibody was also evaluated in the in vitro MLR assay described in Example 9. Again, the parental 11.5.1 antibody inhibited proliferation of humanized T cells because its net effect was agonism of the BTLA receptor. The FcR-null D265A mutation abolished the agonistic effect of the antibody, and thus the antibody showed no effect in this assay ( Figure 10 b) The FcR null 11.5.1 antibody did not enhance proliferation of the humanized cells in this assay because HVEM was blocked (by addition of a polyclonal anti-HVEM antibody), and therefore there was no baseline signaling through the pathway to be blocked by the BTLA blocking antibody.

[0358] Example 14. Antibodies 2.8.6 and 11.5.1 do not fix complement in vitro

[0359] Splenocytes from humanized mice were incubated with 10% baby rabbit complement (BioRad) and anti-BTLA antibody (or isotype control or positive control depleting anti-CD20 antibody; clone SA271G2 from Biolegend) at 20 μg / ml for 15 min at 37°C. The anti-CD20 antibody depletes most of the B220 + B cells, while anti-BTLA antibodies do not deplete B220 + or CD4 + cell( Figure 11 ), even though both groups stained positive for BTLA.

[0360] Example 15. Antibodies 2.8.6 and 11.5.1 do not induce ADCC in vitro

[0361] Whole spleen cells (including myeloid effector cells) from humanized mice were incubated with 20 μg / ml of anti-BTLA antibody (or isotype control or depleting anti-CD20 antibody SA271G2) at 37°C for 24 hours. +cells, while anti-BTLA antibodies did not deplete B220 + or CD4 + cell( Figure 12 ), even though both groups stained positive for BTLA.

[0362] Example 16. Antibodies 2.8.6 and 11.5.1 do not deplete BTLA-expressing cells in vivo

[0363] Humanized BTLA mice were injected intraperitoneally with 200 μg of anti-BTLA antibody or isotype control. Spleens were harvested at 24 hours, and the frequencies of different cell populations were determined by flow cytometry. Anti-BTLA antibody had no effect on the frequency or absolute number of B cells or T cells in the spleen or the number of B cell precursors in the bone marrow ( Figure 13 ).

[0364] Example 17. Antibodies 2.8.6 and 11.5.1 stabilize BTLA expression on immune cells in vivo

[0365] Humanized mice were injected intraperitoneally with 10 mg / kg of antibody 2.8.6 or 11.5.1. Six days after injection, mice were humanely sacrificed, spleens were harvested and processed into single cell suspensions for flow cytometric evaluation. Cells were stained with a cocktail of antibodies to identify immune cell subsets and with a fluorescently conjugated anti-BTLA antibody whose epitope is non-competitive with the injected antibody. The geometric mean of BTLA staining after incubation with anti-BTLA antibody in vivo was normalized to the geometric mean of BTLA staining after incubation with isotype control (using the same staining antibody). BTLA expression on B cells and CD4 T cells from mice injected with clone 2.8.6 or 11.5.1 was significantly higher compared to mice injected with isotype control ( Figure 14 This suggests that clones 2.8.6 and 11.5.1 stabilize BTLA expression on cell surfaces in vivo, rather than inducing receptor downregulation, as observed with other BTLA antibodies in the prior art (M.-L. del Rio et al. / Immunobiology 215 (2010) 570–578). For immunosuppressive purposes, agonist antibodies that stabilize receptor expression have the advantage of being able to prolong high levels of inhibitory signaling through the pathway compared to downregulating antibodies.

[0366] Example 18. Tolerance and side effects in animal models

[0367] No tolerability issues or side effects were observed in any animal studies using antibodies 2.8.6 or 11.5.1.

[0368] Example 19. Humanization of Antibody 2.8.6

[0369] Antibody 2.8.6 was humanized by CDR grafting onto homologous human germline framework regions (see SEQ ID NOs: 13-14). IGHV2-5*08 was used for the heavy chain and IGKV3-11*01 was used for the light chain. After humanization, binding to BTLA was assessed by SPR. Humanized 2.8.6 had a K of 0.73 nM. D Binds to monomeric BTLA.

[0370] Example 20. Characterization of Exemplary BTLA Antibodies

[0371] In addition to 2.8.6 and 11.5.1, the characterization of the exemplary mIgG1 BTLA antibodies provided herein is also described in this example. The binding affinity of the various clones listed in Table 1 to BTLA and the inhibitory efficiency of lymphocytes were evaluated (Table 2). For each antibody, the association rate ("on rate") and dissociation rate ("off rate") for binding to human BTLA, as well as the KD for binding to human or cynomolgus monkey BTLA were measured according to the method described in Example 2, and the injection curve of a single concentration of the BTLA extracellular domain was fitted. The inhibitory efficiency of a single antibody on T cells was also evaluated at a single concentration of 10 μg / ml. Each individual antibody was subjected to an MLR assay according to the method described in Example 9 (two biological replicates, as shown in Table 3); an anti-CD3 assay was performed according to the following method (two biological replicates, Table 3); and the inhibition of NFkB signaling in the Jurkat T cell line transfected with human BTLA by each antibody was determined according to the method described in Example 10 (Table 3). For each exemplary antibody, the mean inhibition of T cells relative to isotype controls in various in vitro stimulation assays was calculated as the average of the percent inhibition across all assays (Tables 2 and 3).

[0372] The ability of BTLA agonist antibodies to inhibit anti-CD3 and anti-CD28-induced T cell activation was assessed as follows. Splenocytes from humanized BTLA mice were processed into single cell suspensions and treated with ACK buffer to lyse red blood cells. Cells were stained with CFSE (Biolegend catalog number 423801) to allow tracking of cell proliferation. 2 x 10 cells were plated per well. 5Cells were seeded in 96-well U-bottom plates containing soluble anti-CD3 antibodies (clone 145.2C11; Biolegend #100339) and anti-CD28 (clone 37.51; Biolegend #102115) at a concentration of 50 ng / ml each, along with soluble anti-BTLA antibodies or isotype controls at a concentration of 10 μg / ml. After 72 hours, cells were analyzed by flow cytometry to assess proliferation ("Anti-CD3 / CD28 (CD4 T Cell Proliferation)") and T cell activation ("Anti-CD3 / CD28 (CD69+CD4 T Cells)") by staining for surface-expressed activation markers. For each BTLA antibody, the percent inhibition was calculated compared to the isotype control antibody.

[0373] In addition, for each BTLA antibody, its ligand blocking ability, for example, the ability to compete with HVEM for BTLA binding was evaluated according to the method described in Example 4, with a result indicated as "yes" for inhibition of HVEM-BTLA binding exceeding 90%, and a result indicated as "no" for inhibition of HVEM-BTLA binding less than 10%. The functional epitope of each BTLA antibody was also determined according to the method described in Example 5. The "Epitope" column in Table 2 summarizes the epitope group bound by each individual BTLA antibody. Antibodies 2.8.6, 6.2, 831, 16H2, 7A1, 16F10, 6G8, 3E8, 4E8, 15C6, 12F11, 10B1, 15B6, 4D3, 16E1, 4D5, and 3A9 all bind to a first epitope (referred to in the table as "epitope 1") comprising at least one key residue selected from the following list: D52, P53, E55, E57, E83, Q86, E103, L106, and E92. Antibodies that bind to epitope 1 do not compete with the ligand HVEM for binding to BTLA. Antibodies 11.5.1, 14D4, 1H6, 8C4, 27G9, and 26F3 all bind to a different second epitope ("epitope 2") comprising at least one key residue selected from the following list: Y39, K41, R42, Q43, E45, and S47. Antibodies binding to epitope 2 compete with the ligand HVEM for BTLA binding. Antibody 26B1 binds to a third epitope ("epitope 3") comprising at least one key residue selected from the following list: D35, T78, K81, S121, and L123. Antibodies binding to epitope 3 compete with the ligand HVEM for BTLA binding. Antibodies 24H7, 4B1, 8B4, and 4H4 all bind to a different fourth epitope ("epitope 4") comprising the key residue H68. Antibodies binding to epitope 4 do not compete with the ligand HVEM for BTLA binding. Antibody 21C7 binds to a different fifth epitope ("Epitope 5") comprising at least one key residue selected from the following list: N65 and A64. Antibodies binding to Epitope 5 do not compete with the ligand HVEM for binding to BTLA.

[0374] Example 21. Humanization and CDR Engineering of BTLA Antibodies 3E8 and 6.2

[0375] The variable domains of 3E8 and 6.2 were humanized by germlining into homologous human germline framework regions (Seq ID No. 382–385). For 3E8, the selected acceptor frameworks were VH1-1-08 and JH6 for the heavy chain and VK3-L6 and JK2 for the light chain. For 6.2, the selected acceptor frameworks were VH3-3-21 and JH6 for the heavy chain and VK2-A19 and JK4 for the light chain.

[0376] Sometimes certain residues in an antibody CDR can be replaced to remove unwanted features without significantly affecting target binding. The CDRH2 of humanized antibody 6.2 was modified with D54E and N56Q substitutions (Seq ID No. 387) to remove deamidation potential (the engineered humanized VH sequence of 6.2 is given in Seq ID No. 390). Similarly, as determined by Lonza's Epibase analysis, the CDRH2 of humanized antibody 3E8 was modified with N57Q substitutions (Seq ID No. 388) to remove deamidation potential and with K63S substitutions to reduce predicted immunogenicity (the engineered humanized VH sequence of 3E8 is given in Seq ID No. 389).

[0377] Table 2. Characterization of Binding Affinity and Inhibition of Exemplary Antibodies

[0378]

[0379]

[0380] Table 3. Inhibition assay results of exemplary antibodies

[0381]

[0382]

[0383] Sequence Listing

[0384] SEQ ID NO: 1

[0385] GDSITSAY

[0386] SEQ ID NO:2

[0387] ISYSGST

[0388] SEQ ID NO:3

[0389] ARSHYYGYYFDY

[0390] SEQ ID NO:4

[0391] ETIDSYGDSL

[0392] SEQ ID NO:5

[0393] RAS

[0394] SEQ ID NO:6

[0395] QQTDEDPYT

[0396] SEQ ID NO:7

[0397] GFSLTTYG

[0398] SEQ ID NO:8

[0399] MWPGGRT

[0400] SEQ ID NO:9

[0401] VRGDYEYDYYAMDY

[0402] SEQ ID NO:10

[0403] SSVSY

[0404] SEQ ID NO:11

[0405] ATS

[0406] SEQ ID NO:12

[0407] HQWSSNPYT

[0408] SEQ ID NO:13 - Humanized antibody 2.8.6 VH region

[0409] QVTLKESGPALVKPTQTLTLTCTVSGFSLTTYGVHWIRQPPGKALEWLGVMWPGGRTSYNPSLKSRLTITKDNSKSQVVLTMTNMDPVDTATYYCVRGDYEYDYYAMDYWGQGTLVTVSS

[0410] SEQ ID NO:14 - Humanized antibody 2.8.6 VL region

[0411] EIVLTQSPATLSLSPGERATLSCRASSSVSYMHWYQQKPGQAPRPLIYATSNRATGIPARFSGSGSGTDYTLTISSLEPEDFAVYYCHQWSSNPYTFGQGTKLEIK

[0412] SEQ ID NO:15 - Mouse Ab 2.8.6 VL region

[0413] QIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPKPWIYATSNLASGVPAR FSGSGSGTSYSLTISRMEAEDAATYYCHQWSSNPYTFGGGTKLEIK

[0414] SEQ ID NO:16

[0415] caaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagttcaagtgtaagttacatgcactggtaccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgctcgcttcagtggcagtgggtctgggacctcttactctctcacaatcagcagaatggaggctgaagatgctgccacttattactgccaccagtggagtagtaacccgtacacgttcggaggggggaccaagctggaaataaaac

[0416] SEQ ID NO:17 = Mouse Ab 2.8.6 VH region

[0417] QVQLKESGPGLVAPSQSLSITCTVSGFSLTTYGVHWVRQSPGKGLEWLGVMWPGGRTSYNPAPMSRLSISKDNSKSQVFLKMNSLQTDDTAMYYCVRGDYEYDYYAMDYWGQGTSVTVSS

[0418] SEQ ID NO:18

[0419] caggtgcagctgaaggagtctggacctggcctggtggcgccctcacagagcctgtccatcacttgcactgtctctgggttttcattaaccacctatggtgtacactgggttcgccagtctccaggaaagggtctggagtggctgggagtaatgtggcctggtggaagaacaagttataatccggctcccatgtccagactgagcatcagcaaagacaactccaagagccaagttttcttaaaaatgaacagtctgcaaactgatgacacggccatgtactactgtgtcagaggggactatgaatacgattactatgctatggactactggggtcaaggaacctcagtcaccgtctcctcag

[0420] SEQ ID NO:19 - Mouse Ab 11.5.1 VL Region

[0421] DIVLTQSPASLAVSLGQRATISCRASETIDSYGDSLMHWYQQKAGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQTDEDPYTFGGGTKLEIK

[0422] SEQ ID NO:20

[0423] gacattgtgctgacccaatctccagcttctttggctgtgtctctagggcagagggccaccatatcctgcagagccagtgaaactattgatagttatggcgatagtttaatgcactggtaccagcagaaagcaggacagccacccaaactcctcatctatcgtgcatccaacctagaatctgggatccctgccaggttcagtggcagtgggtctcggacagacttcaccctcaccattaatcctgtggaggctgatgatgttgcaacctattactgtcagcaaactgatgaggatccgtacacgttcggaggggggaccaagctggaaataaaa

[0424] SEQ ID NO:21 - Mouse Ab 11.5.1 VH Region

[0425] EVQLQESGPSLVKPSQTLSLTCSVTGDSITSAYWNWIRKFPGNKLEYMGYISYSGSTYFNPSLKSRISITRNTSKNQYYLQLNSVTTEDTATYYCARSHYYGYYFDYWGHGTTLTVSS

[0426] SEQ ID NO:22

[0427] gaggtgcagcttcaggagtcaggacctagcctcgtgaaaccttctcagactctgtccctcacctgttctgtcactggcgactccatcaccagtgcttactggaactggatccggaaattcccagggaataaacttgagtacatggggtacataagctacagtggtagcacttacttc aatccatctctcaaaagtcgaatctccatcactcgaaacacatccaagaaccagtactacctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagatctcattactacggctactactttgactactggggccatggcaccactctcacagtctcctca

[0428] SEQ ID NO:23—Human (Homo sapiens) BTLA polypeptide. Positions 1-30 are the signal sequence, 31-151 are the extracellular region, 152-178 are the transmembrane region, and 179 to the end are the intracellular region.

[0429] MKTLPAMLGTGKLFWVFFLIPYLDIWNIHGKESCDVQLYIKRQSEHSILAGDPFELECPVKYCANRPHVTWCKLNGTTCVKLEDRQTSWKEEKNISFFILHFEPVLPNDNGSYRCSANFQ SNLIESHSTTLYVTDVKSASERPSKDEMASRPWLLYRLLPLGPLLITTCFCLFCCLRRHQGKQNELSDTAGREINLVDAHLKSEQTEASTRQNSQVLLSETGIYDNDPDLCFRMQEGS EVYSNPCLEENKPGIVYASLNHSVIGPNSRLARNVKEAPTEYASICVRS

[0430] SEQ ID NO:24—Macaca fascicularis BTLA polypeptide.

[0431] MKTLPAMLGSGRLFWVVFLIPYLDIWNIHGKESCDVQLYIKRQSYHSIFAGDPFKLECPVKYCAHRPQVTWCKLNGTTCVKLEGRHTSWKQEKNLSFFILHFEPVLPSDNGSYRCSANFLSAIIESHSTTLYVTDVKSASERPS KDEMASRPWLLYSLLPLGLPPLLITTCFCLFCFLRRHQGKQNELSDTTGREITLVDVPFKSEQTEASTRQNSQVLLSETGIYDNEPDFCFRMQEGSEVYSNPCLEENKPGIIYASLNHSIIGLNSRQARNVKEAPTEYASICVRS

[0432] Table 4. Exemplary CDR sequences

[0433]

[0434]

[0435]

[0436] Table 5. Exemplary primary VH and VL sequences

[0437]

[0438]

[0439]

[0440] Sequence Listing <110> Oxford University Technology Innovation Co., Ltd. Miro Biological Co., Ltd. <120> Anti-BTLA antibodies <130> P266301WO <150> GB1820554.2 <151> 2018-12-17 <160> 390 <170> PatentIn version 3.5 <210> 1 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 1 Gly Asp Ser Ile Thr Ser Ala Tyr 1 5 <210> 2 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 2 Ile Ser Tyr Ser Gly Ser Thr 1 5 <210> 3 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 3 Ala Arg Ser His Tyr Tyr Gly Tyr Tyr Phe Asp Tyr 1 5 10 <210> 4 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 4 Glu Thr Ile Asp Ser Tyr Gly Asp Ser Leu 1 5 10 <210> 5 <211> 3 <212> PRT <213> Mouse (Mus musculus) <400> 5 Arg Ala Ser 1 <210> 6 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 6 Gln Gln Thr Asp Glu Asp Pro Tyr Thr 1 5 <210> 7 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 7 Gly Phe Ser Leu Thr Thr Tyr Gly 1 5 <210> 8 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 8 Met Trp Pro Gly Gly Arg Thr 1 5 <210> 9 <211> 14 <212> PRT <213> Mouse (Mus musculus) <400> 9 Val Arg Gly Asp Tyr Glu Tyr Asp Tyr Tyr Ala Met Asp Tyr 1 5 10 <210> 10 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 10 Ser Ser Val Ser Tyr 1 5 <210> 11 <211> 3 <212> PRT <213> Mouse (Mus musculus) <400> 11 Ala Thr Ser 1 <210> 12 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 12 His Gln Trp Ser Ser Asn Pro Tyr Thr 1 5 <210> 13 <211> 120 <212> PRT <213> Synthetic Sequence <220> <223> Humanized Antibody 2.8.6 VH Region <400> 13 Gln Val Thr Leu Lys Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Thr Tyr 20 25 30 Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu 35 40 45 Gly Val Met Trp Pro Gly Gly Arg Thr Ser Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Leu Thr Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Val Leu 65 70 75 80 Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Val 85 90 95 Arg Gly Asp Tyr Glu Tyr Asp Tyr Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 14 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Humanized Antibody 2.8.6 VL Region <400> 14 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys His Gln Trp Ser Ser Asn Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 15 <211> 106 <212> PRT <213> Mouse (Mus musculus) <400> 15 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys His Gln Trp Ser Ser Asn Pro Tyr Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 16 <211> 319 <212> DNA <213> Mouse (Mus musculus) <400> 16 caaattgttc tctcccagtc tccagcaatc ctgtctgcat ctccagggga gaaggtcaca 60 atgacttgca gggccagttc aagtgtaagt tacatgcact ggtaccagca gaagccagga 120 tcctccccca aaccctggat ttatgccaca tccaacctgg cttctggagt ccctgctcgc 180 ttcagtggca gtgggtctgg gacctcttac tctctcacaa tcagcagaat ggaggctgaa 240 gatgctgcca cttattactg ccaccagtgg agtagtaacc cgtacacgtt cggagggggg 300 accaagctgg aaataaaac 319 <210> 17 <211> 120 <212> PRT <213> Mouse (Mus musculus) <400> 17 Gln Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Thr Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Met Trp Pro Gly Gly Arg Thr Ser Tyr Asn Pro Ala Pro Met 50 55 60 Ser Arg Leu Ser Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Phe Leu 65 70 75 80 Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Met Tyr Tyr Cys Val 85 90 95 Arg Gly Asp Tyr Glu Tyr Asp Tyr Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 18 <211> 361 <212> DNA <213> Mouse (Mus musculus) <400> 18 caggtgcagc tgaaggagtc tggacctggc ctggtggcgc cctcacagag cctgtccatc 60 acttgcactg tctctgggtt ttcattaacc acctatggtg tacactgggt tcgccagtct 120 ccaggaaagg gtctggagtg gctgggagta atgtggcctg gtggaagaac aagttataat 180 ccggctccca tgtccagact gagcatcagc aaagacaact ccaagagcca agttttctta 240 aaaatgaaca gtctgcaaac tgatgacacg gccatgtact actgtgtcag aggggactat 300 gaatacgatt actatgctat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca 360 g 361 <(210)> 19 <(211)> 111 <(212)> PRT <(213)> Mouse (Mus musculus) <(400)> 19 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Thr Ile Asp Ser Tyr 20 25 30 Gly Asp Ser Leu Met His Trp Tyr Gln Gln Lys Ala Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Thr Asp 85 90 95 Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 20 <211> 333 <212> DNA <213> Mouse (Mus musculus) <400> 20 gacattgtgc tgacccaatc tccagcttct ttggctgtgt ctctagggca gagggccacc 60 atatcctgca gagccagtga aactattgat agttatggcg atagtttaat gcactggtac 120 cagcagaaag caggacagcc acccaaactc ctcatctatc gtgcatccaa cctagaatct 180 gggatccctg ccaggttcag tggcagtggg tctcggacag acttcaccct caccattaat 240 cctgtggagg ctgatgatgt tgcaacctat tactgtcagc aaactgatga ggatccgtac 300 acgttcggag gggggaccaa gctggaaata aaa 333 <210> 21 <211> 118 <212> PRT <213> Mouse (Mus musculus) <400> 21 Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Ala 20 25 30 Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 35 40 45 Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Phe Asn Pro Ser Leu Lys 50 55 60 Ser Arg Ile Ser Ile Thr Arg Asn Thr Ser Lys Asn Gln Tyr Tyr Leu 65 70 75 80 Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95 Arg Ser His Tyr Tyr Gly Tyr Tyr Phe Asp Tyr Trp Gly His Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> twenty two <211> 354 <212> DNA <213> Mouse (Mus musculus) <400> twenty two gaggtgcagc ttcaggagtc aggacctagc ctcgtgaaac cttctcagac tctgtccctc 60 acctgttctg tcactggcga ctccatcacc agtgcttact ggaactggat ccggaaattc 120 ccagggaata aacttgagta catggggtac ataagctaca gtggtagcac ttacttcaat 180 ccatctctca aaagtcgaat ctccatcact cgaaacacat ccaagaacca gtactacctg 240 cagttgaatt ctgtgactac tgaggacaca gccacatatt actgtgcaag atctcattac 300 tacggctact actttgacta ctggggccat ggcaccactc tcacagtctc ctca 354 <210> twenty three <211> 289 <212> PRT <213> Homo sapiens <220> <221> MISC_FEATURE <222> (1)..(30) <223> signal sequence <220> <221> MISC_FEATURE <222> (31)..(151) <223> Extracellular region <220> <221> MISC_FEATURE <222> (152)..(178) <223> transmembrane region <220> <221> MISC_FEATURE <222> (179)..(289) <223> Intracellular region <400> 23 Met Lys Thr Leu Pro Ala Met Leu Gly Thr Gly Lys Leu Phe Trp Val 1 5 10 15 Phe Phe Leu Ile Pro Tyr Leu Asp Ile Trp Asn Ile His Gly Lys Glu 20 25 30 Ser Cys Asp Val Gln Leu Tyr Ile Lys Arg Gln Ser Glu His Ser Ile 35 40 45 Leu Ala Gly Asp Pro Phe Glu Leu Glu Cys Pro Val Lys Tyr Cys Ala 50 55 60 Asn Arg Pro His Val Thr Trp Cys Lys Leu Asn Gly Thr Thr Cys Val 65 70 75 80 Lys Leu Glu Asp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn Ile Ser 85 90 95 Phe Phe Ile Leu His Phe Glu Pro Val Leu Pro Asn Asp Asn Gly Ser 100 105 110 Tyr Arg Cys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser His Ser 115 120 125 Thr Thr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg Pro Ser 130 135 140 Lys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Arg Leu Leu Pro 145 150 155 160 Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys Leu Phe Cys 165 170 175 Cys Leu Arg Arg His Gln Gly Lys Gln Asn Glu Leu Ser Asp Thr Ala 180 185 190 Gly Arg Glu Ile Asn Leu Val Asp Ala His Leu Lys Ser Glu Gln Thr 195 200 205 Glu Ala Ser Thr Arg Gln Asn Ser Gln Val Leu Leu Ser Glu Thr Gly 210 215 220 Ile Tyr Asp Asn Asp Pro Asp Leu Cys Phe Arg Met Gln Glu Gly Ser 225 230 235 240 Glu Val Tyr Ser Asn Pro Cys Leu Glu Glu Asn Lys Pro Gly Ile Val 245 250 255 Tyr Ala Ser Leu Asn His Ser Val Ile Gly Pro Asn Ser Arg Leu Ala 260 265 270 Arg Asn Val Lys Glu Ala Pro Thr Glu Tyr Ala Ser Ile Cys Val Arg 275 280 285 Ser <210> 24 <211> 289 <212> PRT <213> Cynomolgus monkey (Macaca fascicularis) <400> 24 Met Lys Thr Leu Pro Ala Met Leu Gly Ser Gly Arg Leu Phe Trp Val 1 5 10 15 Val Phe Leu Ile Pro Tyr Leu Asp Ile Trp Asn Ile His Gly Lys Glu 20 25 30 Ser Cys Asp Val Gln Leu Tyr Ile Lys Arg Gln Ser Tyr His Ser Ile 35 40 45 Phe Ala Gly Asp Pro Phe Lys Leu Glu Cys Pro Val Lys Tyr Cys Ala 50 55 60 His Arg Pro Gln Val Thr Trp Cys Lys Leu Asn Gly Thr Thr Cys Val 65 70 75 80 Lys Leu Glu Gly Arg His Thr Ser Trp Lys Gln Glu Lys Asn Leu Ser 85 90 95 Phe Phe Ile Leu His Phe Glu Pro Val Leu Pro Ser Asp Asn Gly Ser 100 105 110 Tyr Arg Cys Ser Ala Asn Phe Leu Ser Ala Ile Ile Glu Ser His Ser 115 120 125 Thr Thr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg Pro Ser 130 135 140 Lys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Ser Leu Leu Pro 145 150 155 160 Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys Leu Phe Cys 165 170 175 Phe Leu Arg Arg His Gln Gly Lys Gln Asn Glu Leu Ser Asp Thr Thr 180 185 190 Gly Arg Glu Ile Thr Leu Val Asp Val Pro Phe Lys Ser Glu Gln Thr 195 200 205 Glu Ala Ser Thr Arg Gln Asn Ser Gln Val Leu Leu Ser Glu Thr Gly 210 215 220 Ile Tyr Asp Asn Glu Pro Asp Phe Cys Phe Arg Met Gln Glu Gly Ser 225 230 235 240 Glu Val Tyr Ser Asn Pro Cys Leu Glu Glu Asn Lys Pro Gly Ile Ile 245 250 255 Tyr Ala Ser Leu Asn His Ser Ile Ile Gly Leu Asn Ser Arg Gln Ala 260 265 270 Arg Asn Val Lys Glu Ala Pro Thr Glu Tyr Ala Ser Ile Cys Val Arg 275 280 285 Ser <210> 25 <400> 25 000 <210> 26 <400> 26 000 <210> 27 <400> 27 000 <210> 28 <400> 28 000 <210> 29 <400> 29 000 <210> 30 <400> 30 000 <210> 31 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 31 Ser Tyr Gly Ile Ser 1 5 <210> 32 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 32 Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 33 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 33 Asn Tyr Gly Ser Ser Tyr Pro Phe Ala Tyr 1 5 10 <210> 34 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 34 Ser Ala Ser Ser Ser Val Ser Ser Ser Tyr Leu His 1 5 10 <210> 35 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 35 Arg Thr Ser Asn Leu Ala Ser 1 5 <210> 36 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 36 Gln Gln Trp Ser Gly Tyr Pro Phe Thr 1 5 <210> 37 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 37 Asp Tyr Tyr Met Asn 1 5 <210> 38 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 38 Asp Ile Asn Pro Asn Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 39 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 39 Trp Arg Gln Leu Arg Ser Asp Tyr 1 5 <210> 40 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 40 Leu Ala Ser Gln Thr Ile Gly Thr Trp Leu Ala 1 5 10 <210> 41 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 41 Ala Ala Thr Ser Leu Ala Asp 1 5 <210> 42 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 42 Gln Gln Leu Tyr Ser Thr Pro Leu Thr 1 5 <210> 43 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 43 Ser Tyr Trp Met His 1 5 <210> 44 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 44 Met Ile His Pro Asn Asn Gly Ile Pro Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 45 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 45 Glu Gly Tyr Tyr Gly Ser Glu Gly Tyr Phe Asp Val 1 5 10 <210> 46 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 46 Ser Ala Ser Ser Ser Ser Ile Ser Tyr Ile His 1 5 10 <210> 47 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 47 Asp Thr Ser Lys Leu Ala Ser 1 5 <210> 48 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 48 His Gln Arg Ser Thr Tyr Pro Tyr Thr 1 5 <210> 49 <400> 49 000 <210> 50 <400> 50 000 <210> 51 <400> 51 000 <210> 52 <400> 52 000 <210> 53 <400> 53 000 <210> 54 <400> 54 000 <210> 55 <400> 55 000 <210> 56 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 56 Met Ile His Pro Asn Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 57 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 57 Lys Arg Gly Gly Leu Gly Asp Tyr 1 5 <210> 58 <211> 15 <212> PRT <213> Mouse (Mus musculus) <400> 58 Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Tyr Met His 1 5 10 15 <210> 59 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 59 Leu Ala Ser Asn Leu Glu Ser 1 5 <210> 60 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 60 Gln His Ser Arg Glu Leu Pro Tyr Thr 1 5 <210> 61 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 61 Ser Ser Trp Met Asn 1 5 <210> 62 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 62 Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys 1 5 10 15 Gly <210> 63 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 63 Arg Gly Tyr Gly Tyr Leu Ala Tyr 1 5 <210> 64 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 64 Lys Ala Ser Gln Asp Val Ser Thr Ala Val Ala 1 5 10 <210> 65 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 65 Ser Ala Ser Tyr Arg Tyr Thr 1 5 <210> 66 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 66 Gln Gln His Tyr Ser Thr Pro Tyr Thr 1 5 <210> 67 <400> 67 000 <210> 68 <400> 68 000 <210> 69 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 69 Gly Tyr Gly Ser Ser Tyr Gly Phe Ala Tyr 1 5 10 <210> 70 <400> 70 000 <210> 71 <400> 71 000 <210> 72 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 72 Gln Gln Trp Ser Gly Tyr Pro Trp Thr 1 5 <210> 73 <211> 6 <212> PRT <213> Mouse (Mus musculus) <400> 73 Ser Gly Tyr Tyr Trp Asn 1 5 <210> 74 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 74 Tyr Ile Ser Tyr Asp Gly Ser Asn Asn Tyr Asn Pro Ser Leu Lys Asn 1 5 10 15 <210> 75 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 75 Ile Tyr Gly Asn Tyr Tyr Ala Met Asp Tyr 1 5 10 <210> 76 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 76 Ser Ala Ser Ser Ser Val Ser Tyr Met His 1 5 10 <210> 77 <400> 77 000 <210> 78 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 78 Gln Gln Trp Ser Ser Asn Pro Pro Thr 1 5 <210> 79 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 79 Asp Tyr Tyr Met Ile 1 5 <210> 80 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 80 Asn Ile Asn Pro Asn Asn Gly Gly Thr Thr Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 81 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 81 Gly Gly Leu Arg Pro Leu Tyr Phe Asp Tyr 1 5 10 <210> 82 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 82 Lys Ala Ser Glu Asn Val Asp Thr Tyr Val Ser 1 5 10 <210> 83 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 83 Gly Ala Ser Asn Arg Tyr Thr 1 5 <210> 84 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 84 Gly Gln Ser Tyr Ser Tyr Pro Leu Thr 1 5 <210> 85 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 85 Asn Thr Tyr Met His 1 5 <210> 86 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 86 Arg Ile Asp Pro Ala Asn Gly Asn Thr Lys Tyr Asp Pro Lys Phe Gln 1 5 10 15 Gly <210> 87 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 87 Thr Tyr Tyr Gly Ser Ser Gln His Tyr Phe Asp Tyr 1 5 10 <210> 88 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 88 Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn 1 5 10 15 <210> 89 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 89 Leu Val Ser Lys Leu Asp Ser 1 5 <210> 90 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 90 Trp Gln Asp Thr His Phe Pro Gln Thr 1 5 <210> 91 <400> 91 000 <210> 92 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 92 Arg Ile Tyr Pro Gly Asp Gly Asp Ala Asn Tyr Asn Gly Lys Phe Lys 1 5 10 15 Gly <210> 93 <211> 15 <212> PRT <213> Mouse (Mus musculus) <400> 93 Glu Gly His Tyr Tyr Gly Ser Gly Tyr Arg Trp Tyr Leu Asp Val 1 5 10 15 <210> 94 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 94 Arg Ala Ser Glu Asn Ile Tyr Ser Asn Leu Ala 1 5 10 <210> 95 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 95 Ala Ala Thr Asn Leu Ala Asp 1 5 <210> 96 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 96 Gln His Phe Arg Gly Ala Pro Phe Thr 1 5 <210> 97 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 97 Asp Tyr Glu Ile His 1 5 <210> 98 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 98 Pro Ile Asp Pro Asp Thr Gly Asn Thr Ala Tyr Asn Gln Asn Leu Lys 1 5 10 15 Gly <210> 99 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 99 Gly Gly Tyr Asp Ser Asp Trp Gly Phe Ala Tyr 1 5 10 <210> 100 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 100 Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Phe Leu Phe 1 5 10 15 <210> 101 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 101 Arg Met Ser Asp Leu Ala Ser 1 5 <210> 102 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 102 Met Gln His Leu Glu Tyr Pro Phe Thr 1 5 <210> 103 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 103 Asp Tyr Tyr Leu Asn 1 5 <210> 104 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 104 Leu Ile Asp Pro Tyr Asn Gly Gly Ser Ser Cys Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 105 <211> 6 <212> PRT <213> Mouse (Mus musculus) <400> 105 Gly Asn Ala Met Asp Tyr 1 5 <210> 106 <400> 106 000 <210> 107 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 107 Trp Ala Ser Thr Arg His Thr 1 5 <210> 108 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 108 Gln Gln His Tyr Ile Ile Pro Tyr Met 1 5 <210> 109 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 109 Asn Thr Tyr Met Tyr 1 5 <210> 110 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 110 Arg Ile Asp Pro Ala Asn Gly Asn Thr Lys Tyr Ala Pro Lys Phe Gln 1 5 10 15 Gly <210> 111 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 111 Leu Tyr Tyr Gly Ser Ser Tyr Asp Tyr Phe Asp Tyr 1 5 10 <210> 112 <400> 112 000 <210> 113 <400> 113 000 <210> 114 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 114 Trp Gln Gly Thr His Phe Pro Gln Thr 1 5 <210> 115 <400> 115 000 <210> 116 <400> 116 000 <210> 117 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 117 Thr Tyr Tyr Gly Ser Ser Gln Tyr Tyr Phe Asp Tyr 1 5 10 <210> 118 <400> 118 000 <210> 119 <400> 119 000 <210> 120 <400> 120 000 <210> 121 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 121 Asp Tyr Tyr Ile Asn 1 5 <210> 122 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 122 Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 123 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 123 Gly Tyr Gly Asn Ser Asp Tyr 1 5 <210> 124 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 124 Arg Ala Ser Gln Ser Ile Gly Thr Arg Ile His 1 5 10 <210> 125 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 125 Tyr Ala Ser Glu Ser Ile Ser 1 5 <210> 126 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 126 Gln Gln Ser Asn Ser Trp Pro Tyr Thr 1 5 <210> 127 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 127 Ser Tyr Ala Ile Arg 1 5 <210> 128 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 128 Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 129 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 129 Ser Gly Gly Ala Ser Tyr Thr Met Asp Tyr 1 5 10 <210> 130 <400> 130 000 <210> 131 <400> 131 000 <210> 132 <400> 132 000 <210> 133 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 133 Ser Tyr Gly Leu Ile 1 5 <210> 134 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 134 Glu Ile Tyr Pro Arg Ser Gly Ser Thr Tyr Tyr Tyr Asn Glu Trp Phe Lys 1 5 10 15 Gly <210> 135 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 135 Arg Arg Gly Thr Gly Asp Gly Phe Asp Tyr 1 5 10 <210> 136 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 136 Ser Ala Ser Gln Gly Ile Ser Asn Tyr Leu Asn 1 5 10 <210> 137 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 137 Tyr Thr Ser Ser Leu His Ser 1 5 <210> 138 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 138 Gln Gln Tyr Ile Glu Leu Pro Phe Thr 1 5 <210> 139 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 139 Asp Tyr Tyr Met His 1 5 <210> 140 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 140 Tyr Ile Tyr Pro Asn Asn Gly Gly Asn Gly Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 141 <211> 13 <212> PRT <213> Mouse (Mus musculus) <400> 141 Gly Asp Tyr Tyr Gly Ser Leu Arg Leu Thr Phe Ala Tyr 1 5 10 <210> 142 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 142 Lys Ser Ser Gln Ser Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu 1 5 10 15 Ala <210> 143 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 143 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 144 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 144 Gln Gln Tyr Tyr Ser Tyr Pro Leu Thr 1 5 <210> 145 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 145 Thr Tyr Gly Val Ser 1 5 <210> 146 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 146 Trp Ile Asn Thr Tyr Ser Gly Val Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 147 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 147 Val Thr Thr Ile Leu His Trp Tyr Phe Asp Val 1 5 10 <210> 148 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 148 Arg Ala Ser Gln Glu Ile Ser Gly Tyr Leu Ser 1 5 10 <210> 149 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 149 Ala Ala Ser Thr Leu Asp Ser 1 5 <210> 150 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 150 Leu Gln Tyr Ala Ser Tyr Pro Phe Thr 1 5 <210> 151 <400> 151 000 <210> 152 <400> 152 000 <210> 153 <400> 153 000 <210> 154 <400> 154 000 <210> 155 <400> 155 000 <210> 156 <400> 156 000 <210> 157 <400> 157 000 <210> 158 <400> 158 000 <210> 159 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 159 Arg Arg Gly Ala Gly Asp Gly Phe Asp Tyr 1 5 10 <210> 160 <400> 160 000 <210> 161 <400> 161 000 <210> 162 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 162 Gln Gln Tyr Ser Lys Leu Pro Phe Thr 1 5 <210> 163 <400> 163 000 <210> 164 <400> 164 000 <210> 165 <400> 165 000 <210> 166 <400> 166 000 <210> 167 <400> 167 000 <210> 168 <400> 168 000 <210> 169 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 169 Asp His Thr Ile His 1 5 <210> 170 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 170 Tyr Ile Tyr Pro Arg Asp Gly Ser Thr Lys Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 171 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 171 Ser Asn Trp Asn Phe Asp Tyr 1 5 <210> 172 <211> 11 <212> PRT <213> Mouse (Mus musculus) <400> 172 Lys Ala Ser Gln Asp Val Gly Thr Ala Val Ala 1 5 10 <210> 173 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 173 Trp Ala Ser Thr Arg Arg Thr 1 5 <210> 174 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 174 Gln Gln Tyr Ser Ser Tyr Pro Leu Thr 1 5 <210> 175 <400> 175 000 <210> 176 <400> 176 000 <210> 177 <400> 177 000 <210> 178 <400> 178 000 <210> 179 <400> 179 000 <210> 180 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 180 Gln Gln His Tyr Ser Thr Pro Trp Thr 1 5 <210> 181 <400> 181 000 <210> 182 <211> 17 <212> PRT <213> Mouse (Mus musculus) <400> 182 Glu Ile Tyr Pro Arg Ser Gly Thr Thr Tyr Tyr Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 183 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 183 Arg Ile Ser Ser Gly Ser Gly Val Asp Tyr 1 5 10 <210> 184 <400> 184 000 <210> 185 <400> 185 000 <210> 186 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 186 Gln Gln Tyr Ser Glu Leu Pro Trp Thr 1 5 <210> 187 <211> 6 <212> PRT <213> Mouse (Mus musculus) <400> 187 Ser Gly Tyr Asp Trp His 1 5 <210> 188 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 188 Tyr Ile Ser Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 189 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 189 Gly Thr Pro Val Val Ala Glu Asp Tyr Phe Asp Tyr 1 5 10 <210> 190 <211> 14 <212> PRT <213> Mouse (Mus musculus) <400> 190 Arg Ser Ser Thr Gly Ala Val Thr Thr Ser Asn Tyr Ala Asn 1 5 10 <210> 191 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 191 Ala Thr Asn Asn Arg Ala Pro 1 5 <210> 192 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 192 Ala Leu Trp Tyr Ser Asn His Leu Val 1 5 <210> 193 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 193 Thr Tyr Gly Val His 1 5 <210> 194 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 194 Val Met Trp Pro Gly Gly Arg Thr Ser Tyr Asn Pro Ala Pro Met Ser 1 5 10 15 <210> 195 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 195 Gly Asp Tyr Glu Tyr Asp Tyr Tyr Ala Met Asp Tyr 1 5 10 <210> 196 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 196 Arg Ala Ser Ser Ser Val Ser Tyr Met His 1 5 10 <210> 197 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 197 Ala Thr Ser Asn Leu Ala Ser 1 5 <210> 198 <400> 198 000 <210> 199 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 199 Ser Ala Tyr Trp Asn 1 5 <210> 200 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 200 Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Phe Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 201 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 201 Ser His Tyr Tyr Gly Tyr Tyr Phe Asp Tyr 1 5 10 <210> 202 <211> 15 <212> PRT <213> Mouse (Mus musculus) <400> 202 Arg Ala Ser Glu Thr Ile Asp Ser Tyr Gly Asp Ser Leu Met His 1 5 10 15 <210> 203 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 203 Arg Ala Ser Asn Leu Glu Ser 1 5 <210> 204 <400> 204 000 <210> 205 <211> 5 <212> PRT <213> Mouse (Mus musculus) <400> 205 Ser Tyr Gly Met Ser 1 5 <210> 206 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 206 Ser Ile Arg Ser Asp Gly Asn Thr Tyr Tyr Pro Asp Ser Val Lys Gly 1 5 10 15 <210> 207 <211> 10 <212> PRT <213> Mouse (Mus musculus) <400> 207 Gly Gly Tyr Tyr Gly Ser Ser Pro Tyr Tyr 1 5 10 <210> 208 <400> 208 000 <210> 209 <211> 7 <212> PRT <213> Mouse (Mus musculus) <400> 209 Trp Ala Ser Thr Arg Asp Ser 1 5 <210> 210 <211> 8 <212> PRT <213> Mouse (Mus musculus) <400> 210 Gln Gln Tyr Tyr Asn Tyr Leu Thr 1 5 <210> 211 <211> 6 <212> PRT <213> Mouse (Mus musculus) <400> 211 Ser Gly Tyr Ser Trp His 1 5 <210> 212 <211> 16 <212> PRT <213> Mouse (Mus musculus) <400> 212 Tyr Ile His Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 213 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 213 Gly Pro His Arg Tyr Asp Gly Val Trp Phe Ala Tyr 1 5 10 <210> 214 <211> 12 <212> PRT <213> Mouse (Mus musculus) <400> 214 Ser Ala Ser Ser Ser Ile Ser Ser Asn Tyr Leu His 1 5 10 <210> 215 <400> 215 000 <210> 216 <211> 9 <212> PRT <213> Mouse (Mus musculus) <400> 216 Gln Gln Gly Thr Asn Ile Pro Leu Thr 1 5 <210> 217 <400> 217 000 <210> 218 <400> 218 000 <210> 219 <400> 219 000 <210> 220 <400> 220 000 <210> 221 <400> 221 000 <210> 222 <400> 222 000 <210> 223 <400> 223 000 <210> 224 <400> 224 000 <210> 225 <400> 225 000 <210> 226 <400> 226 000 <210> 227 <400> 227 000 <210> 228 <400> 228 000 <210> 229 <400> 229 000 <210> 230 <400> 230 000 <210> 231 <400> 231 000 <210> 232 <400> 232 000 <210> 233 <400> 233 000 <210> 234 <400> 234 000 <210> 235 <400> 235 000 <210> 236 <400> 236 000 <210> 237 <400> 237 000 <210> 238 <400> 238 000 <210> 239 <400> 239 000 <210> 240 <400> 240 000 <210> 241 <400> 241 000 <210> 242 <400> 242 000 <210> 243 <400> 243 000 <210> 244 <400> 244 000 <210> 245 <400> 245 000 <210> 246 <400> 246 000 <210> 247 <400> 247 000 <210> 248 <400> 248 000 <210> 249 <400> 249 000 <210> 250 <400> 250 000 <210> 251 <400> 251 000 <210> 252 <400> 252 000 <210> 253 <400> 253 000 <210> 254 <400> 254 000 <210> 255 <400> 255 000 <210> 256 <400> 256 000 <210> 257 <400> 257 000 <210> 258 <400> 258 000 <210> 259 <400> 259 000 <210> 260 <400> 260 000 <210> 261 <400> 261 000 <210> 262 <400> 262 000 <210> 263 <400> 263 000 <210> 264 <400> 264 000 <210> 265 <400> 265 000 <210> 266 <400> 266 000 <210> 267 <400> 267 000 <210> 268 <400> 268 000 <210> 269 <400> 269 000 <210> 270 <400> 270 000 <210> 271 <400> 271 000 <210> 272 <400> 272 000 <210> 273 <400> 273 000 <210> 274 <400> 274 000 <210> 275 <400> 275 000 <210> 276 <400> 276 000 <210> 277 <400> 277 000 <210> 278 <400> 278 000 <210> 279 <400> 279 000 <210> 280 <400> 280 000 <210> 281 <400> 281 000 <210> 282 <400> 282 000 <210> 283 <400> 283 000 <210> 284 <400> 284 000 <210> 285 <400> 285 000 <210> 286 <400> 286 000 <210> 287 <400> 287 000 <210> 288 <400> 288 000 <210> 289 <400> 289 000 <210> 290 <400> 290 000 <210> 291 <400> 291 000 <210> 292 <400> 292 000 <210> 293 <400> 293 000 <210> 294 <400> 294 000 <210> 295 <400> 295 000 <210> 296 <400> 296 000 <210> 297 <400> 297 000 <210> 298 <400> 298 000 <210> 299 <400> 299 000 <210> 300 <400> 300 000 <210> 301 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 301 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Asn Tyr Gly Ser Ser Tyr Pro Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala 115 <210> 302 <211> 117 <212> PRT <213> Mouse (Mus musculus) <400> 302 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Asn Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Asp Ile Asn Pro Asn Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Arg Gln Leu Arg Ser Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Leu Thr Val Ser Ser 115 <210> 303 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 303 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Arg Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile His Pro Asn Asn Gly Ile Pro Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Thr Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr His Cys 85 90 95 Ala Arg Glu Gly Tyr Tyr Gly Ser Glu Gly Tyr Phe Asp Val Trp Gly 100 105 110 Thr Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 304 <400> 304 000 <210> 305 <211> 117 <212> PRT <213> Mouse (Mus musculus) <400> 305 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile His Pro Asn Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70​​​​​​ 100 105 110 Val Thr Val Ser Ser 115 <210> 306 <211> 117 <212> PRT <213> Mouse (Mus musculus) <400> 306 Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Lys Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Gly Tyr Gly Tyr Leu Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 307 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 307 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Gly Tyr Gly Ser Ser Tyr Gly Phe Ala Tyr Trp Gly Gln Gly<000290​​​​​​​​​​​​​​​​​ 1 5 10 15 Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Gly 20 25 30 Tyr Tyr Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Asp Gly Ser Asn Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Asn Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Lys Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Ser Ile Tyr Gly Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 309 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 309 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Gln Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Ile Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Asn Ile Asn Pro Asn Asn Gly Gly Thr Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gly Leu Pro Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Leu Arg Pro Leu Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Leu Thr Val Ser Ser 115 <210> 310 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 310 Glu Val Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asn Thr 20 25 30 Tyr Met His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ala Asn Gly Asn Thr Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Val Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Leu Thr Tyr Tyr Gly Ser Ser Gln His Tyr Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 311 <211> 124 <212> PRT <213> Mouse (Mus musculus) <400> 311 Gln Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Lys Lys Arg Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Asp Ala Asn Tyr Asn Gly Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Gly Glu Gly His Tyr Tyr Gly Ser Gly Tyr Arg Trp Tyr Leu Asp 100 105 110 Val Trp Gly Thr Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 312 <211> 120 <212> PRT <213> Mouse (Mus musculus) <400> 312 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Ile His Trp Val Lys Gln Thr Leu Val His Gly Leu Glu Trp Ile 35 40 45 Gly Pro Ile Asp Pro Asp Thr Gly Asn Thr Ala Tyr Asn Gln Asn Leu 50 55 60 Lys Gly Lys Ala Ile Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Gly Tyr Asp Ser Asp Trp Gly Phe Ala Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 313 <211> 115 <212> PRT <213> Mouse (Mus musculus) <400> 313 Glu Val Gln Leu Gln Gln Ser Gly Pro Val Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Leu Asn Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Leu Ile Asp Pro Tyr Asn Gly Gly Ser Ser Cys Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Asp Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asn Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr 100 105 110 Val Ser Ser 115 <210> 314 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 314 Glu Val Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asn Thr 20 25 30 Tyr Met Tyr Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ala Asn Gly Asn Thr Lys Tyr Ala Pro Lys Phe 50 55 60[[ID=३३]] Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Leu Leu Tyr Tyr Gly Ser Ser Tyr Asp Tyr Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 315 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 315 Glu Val Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asn Thr 20 25 30 Tyr Met His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ala Asn Gly Asn Thr Lys Tyr Ala Pro Lys Phe 50 55 60 Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 [[ID=3৬]] Ala Leu Thr Tyr Tyr Gly Ser Ser Gln Tyr Tyr Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 316 <211> 116 <212> PRT <213> Mouse (Mus musculus) <400> 316 Gln Val Gln Leu Lys Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Glu Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Gly Tyr Gly Asn Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu 100 105 110 Thr Val Ser Ser 115 <210> 317 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 317 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Ile Arg Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Ser Gly Gly Ala Ser Tyr Thr Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 318 ​​​​​​​​​​​​​​​​Gly Leu Ile Trp Leu Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Ser Thr Tyr Tyr Asn Glu Trp Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Phe 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Arg Gly Thr Gly Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ile Leu Thr Val Ser Ser 115 <210> 319 <211> 122 <212> PRT <213> Mouse (Mus musculus) <400> 319 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Asn Asn Gly Gly Asn Gly Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Ile Gly Asp Tyr Tyr Gly Ser Leu Arg Leu Thr Phe Ala Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 320 <211> 120 <212> PRT <213> Mouse (Mus musculus) <400> 320 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Gly Val Ser Trp Val Lys Gln Ala Pro Gly Lys Val Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Ser Gly Val Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Pro Val Thr Thr Ile Leu His Trp Tyr Phe Asp Val Trp Gly Thr 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 321 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 321 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Asn Tyr Gly Ser Ser Tyr Pro Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala 115 <210> 322 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 322 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Arg Gly Ala Gly Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Leu Thr Val Ser Ser 115 <210> 323 <400> 323 000 <210> 324 <211> 116<000​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​100 105 110 Thr Val Ser Ser 115 <210> 325 <400> 325 000 <210> 326 <211> 119 <212> PRT <213> Mouse (Mus musculus) <400> 326 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Pro Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Arg Ser Gly Thr Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80<x Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Ile Ser Ser Gly Ser Gly Val Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Leu Thr Val Ser Ser 1 n1 <210> 327 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 327 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Met Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly 20 2​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​<210> 330 <211> 118 <212> PRT <213> Mouse (Mus musculus) <400> 330 Glu Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Ser Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ile Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Ser Ile Arg Ser Asp Gly Asn Thr Tyr Tyr Pro Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala Arg Asn Ile Leu Tyr Leu 65 70 75 80 Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys Thr 85 90 95 Arg Gly Gly Tyr Tyr Gly Ser Ser Pro Tyr Tyr Trp Gly Gln Gly Thr[[ID=o35]] 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 331 <211> 121 <212> PRT <213> Mouse (Mus musculus) <400> 331 Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Val Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly 20 25 30 Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile His Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Ser Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Ser Gly Pro His Arg Tyr Asp Gly Val Trp Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 332 <400> 332 000 <210> 333 <400> 333 000 <210> 334 <400> 334 000 <210> 335 <400> 335 000 <210> 336 <400> 336 000 <210> 337 <400> 337 000 <210> 338 <400> 338 000 <210> 339 <400> 339 000 <210> 340 <400> 340 000 <210> 341 <400> 341 000 <210> 342 <400> 342 000 <210> 343 <400> 343 000 <210> 344 <400> 344 000 <210> 345 <400> 345 000 <210> 346 <400> 346 000 <210> 347 <400> 347 000 <210> 348 <400> 348 000 <210> 349 <400> 349 000 <210> 350 <400> 350 000 <210> 351 <211> 108 <212> PRT <213> Mouse (Mus musculus) <400> 351 Glu Asn Val Leu Thr Gln Ser Pro Ala Ile Met Ala Ala Ser Leu Gly 1 5 10 15 Gln Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Ser Gly Ala Ser Pro Lys Pro Leu 35 40 45 Ile His Arg Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Val Glu 65 70 75 80 Ala Glu Asp Asp Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Gly Tyr Pro 85 90 95 Phe Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 352 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 352 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Gln Ser Ala Ser Leu Gly 1 5 1​​​​ Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Ala Ala Thr Ser Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Lys Phe Ser Phe Lys Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Phe Val Ser Tyr Tyr Cys Gln Gln Leu Tyr Ser Thr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 353 <211> 106 <212> PRT <213> Mouse (Mus musculus) <400> 353 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Ile Ser Tyr Ile 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys His Gln Arg Ser Thr Tyr Pro Tyr Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 354 <400> 354 000 <210> 355 <211> 111 <212> PRT <213> Mouse (Mus musculus) <400> 355 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys Ser Val Ser Thr Ser 20 25 30 Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala [[ID=​​​​​Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg 85 90 95 Glu Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 356 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 356 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 357 <211> 108 <212> PRT <213> Mouse (Mus musculus) <400> 357 Glu Asn Val Leu Thr Gln Ser Pro Ala Ile Met Ala Ala Ser Leu Gly 1 5 10 15 Gln Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Ser Gly Ala Ser Pro Lys Pro Leu 35 40 45 Ile His Arg Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser s 50 55 60 Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Val Glu 65 70 75 80 Ala Glu Asp Asp Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Gly Tyr Pro 85 90 95 Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 358 <211> 106 <212> PRT <213> Mouse (Mus musculus) <400> 358 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95 Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 359 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 359 Asn Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly 1 5 10 15 Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val Asp Thr Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Glu Gln Ser Pro Lys Leu Leu Ile [[ID=​Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Asp Tyr His Cys Gly Gln Ser Tyr Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Ile 100 105 <210> 360 <211> 112 <212> PRT <213> Mouse (Mus musculus) <400> 360 Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 20 25 30 Asp Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 50 55 60 Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Asp 85 90 95 Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 361 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 361 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Val Gly 1 5 10 15[[ID=二十一]] [[ID=二十二]]Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Arg Gly Ala Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 362 <211> 112 <212> PRT <213> Mouse (Mus musculus) <400> 362 Asp Ile Val Met Thr Gln Ala Thr Pro Ser Val Pro Val Thr Pro Gly 1 5 10 15 Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Asn Thr Phe Leu Phe Trp Phe Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Arg Met Ser Asp Leu Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Met Gln His 85 90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 363[[ID=3F]] <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 363 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Glu Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ile Leu Asn Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Leu Tyr Tyr Cys Gln Gln His Tyr Ile Ile Pro Tyr 85 90 95 Met Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 364 <211> 112 <212> PRT <213> Mouse (Mus musculus) <400> 364 Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 20 25 30 Asp Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 50 55 60 Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly 85 90 95 Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 365 <400> 365 000 <210> 366 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 366 Asp Ile Leu Leu Thr Gln Ser Pro Ala Ile Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Arg 20 25 30 Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser 65 70 75 80 Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Ser Asn Ser Trp Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 367 <211> 108 <212> PRT <213> Mouse (Mus musculus) <400> 367 Glu Asn Val Leu Thr Gln Ser Pro Ala Ile Met Ala Ala Ser Leu Gly 1 5 10 15 Gln Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Ser Ser 20 25 30[[ID=……]](后续内容省略,因为原始文本中未完整给出后续标签及内容)​​​​​​​​​​​​​​​​Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 368 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 368 Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Ile Glu Leu Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 369 <211> 113 <212> PRT <213> Mouse (Mus musculus) <400> 369 Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 370 <211> 107 <212> PRT <213> mice (Mus musculus) <400> 370 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Leu Gln Gln Lys Pro Asp Gly Thr Ile Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Lys Arg Phe Arg Gly 50 55 60 Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser 65 70 75 80 Glu Asp Phe Ala Asp Tyr Tyr Cys Leu Gln Tyr Ala Ser Tyr Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 371 <400> 371 000 <210> 372 <211> 107 <212> PRT <213> mice (Mus musculus) <400> 372 Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Lys Leu Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 373 <400> 373 000 <210> 374 <211> 107 <212> PRT <213> Mouse (Mus musculus) <400> 374 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg Arg Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 375 <211> 107 <2i2> PRT <213> Mouse (Mus musculus) <400> 375 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Ala 20 25 30[[ID=..29]] Val Ala Trp Tyr Gln Gln Lys Pro Gly Gl...

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to B and T lymphocyte attenuator (BTLA), wherein the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, wherein: (i) the heavy chain comprises a heavy chain variable region comprising the following three CDRs: CDRH1, CDRH2, and CDRH3, wherein (a) CDRH1 consists of the amino acid sequence shown in SEQ ID NO: 205; (b) CDRH2 consists of the amino acid sequence set forth in SEQ ID NO: 387; and (c) CDRH3 consists of the amino acid sequence set forth in SEQ ID NO: 207; and (ii) the light chain comprises a light chain variable region comprising the following three CDRs: CDRL1, CDRL2 and CDRL3, wherein (a) CDRL1 consists of the amino acid sequence shown in SEQ ID NO: 142; (b) CDRL2 consists of the amino acid sequence shown in SEQ ID NO: 143; and (c) CDRL3 consists of the amino acid sequence shown in SEQ ID NO:

210.

2. The antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable region comprises an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:

390.

3. The antibody or antigen-binding fragment thereof according to claim 2, wherein the light chain variable region comprises an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:

378.

4. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof binds to a BTLA residue selected from the group consisting of D52, P53, E55, E57, E83, Q86, E103, L106 and E92. The antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a humanized antibody. The antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody.

7. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof agonizes human BTLA expressed on the surface of an immune cell, wherein the immune cell is optionally a T cell.

8. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises a domain that binds to an Fc receptor.

9. The antibody or antigen-binding fragment thereof according to claim 8, wherein the Fc receptor is FcγRIIB.

10. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof specifically binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 10 nM as determined by surface plasmon resonance (SPR) at 37°C.

11. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof binds to cynomolgus monkey BTLA with a KD of less than 20 nM as determined by surface plasmon resonance (SPR) at 37°C.

12. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof does not inhibit the binding of BTLA to herpes virus entry mediator (HVEM).

13. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof inhibits T cell proliferation in vitro as determined by a mixed lymphocyte reaction assay.

14. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof binds to human B and T lymphocyte attenuator (BTLA) with a KD of less than 2 nM as determined by surface plasmon resonance (SPR) at 37°C.

15. The antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:

378.

16. An isolated nucleic acid comprising one or more nucleotide sequences encoding a polypeptide capable of forming the antibody or antigen-binding fragment thereof of claim 1.

17. A host cell comprising one or more nucleic acid molecules encoding a heavy chain amino acid sequence and a light chain amino acid sequence, which, when expressed, can form the antibody or antigen-binding fragment thereof of claim 1.

18. A method of producing the antibody or antigen-binding fragment thereof of claim 1, comprising culturing the host cell of claim 17 under conditions for producing the antibody or antigen-binding fragment thereof.

19. A method for producing the antibody or antigen-binding fragment thereof according to claim 1, the method comprising: a) providing a host cell comprising one or more nucleic acid molecules encoding a heavy chain amino acid sequence and a light chain amino acid sequence, wherein the heavy chain amino acid sequence and the light chain amino acid sequence are capable of forming the antibody or antigen-binding fragment thereof of claim 1 when expressed; b) cultivating a host cell that expresses the encoded amino acid sequence; and c) isolating the antibody or antigen-binding fragment thereof.

20. A pharmaceutical composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof according to claim 1 and at least one pharmaceutically acceptable excipient.

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