Therapeutic use of Anti-tcr delta variable 1 antibodies

Abstract

To provide an anti-Vδ1 antibody or a fragment thereof for use in a method of treating cancer, an infectious disease, or an inflammatory disease.SOLUTION: An anti-Vδ1 antibody or a fragment thereof comprising one or more of a CDR3 comprising a sequence having at least 80% sequence identity with any one of 24 specific amino acid sequences, a CDR2 comprising a sequence having at least 80% sequence identity with any one of 12 specific amino acid sequences, and / or a CDR1 comprising a sequence having at least 80% sequence identity with any one of other 24 specific amino acid sequences.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to the therapeutic use of antibodies and fragments thereof directed against the T cell receptor of gamma delta T cells. [Background technology]

[0002] Growing interest in T cell immunotherapy for cancer has focused on the apparent ability of subsets of CD8+ and CD4+ alpha beta (αβ) T cells to recognize cancer cells and mediate potential host protective functions, particularly when derepressed by clinically mediated antagonism of inhibitory pathways exerted by PD-1, CTLA-4, and other receptors. However, αβ T cells are MHC-restricted, which can lead to graft-versus-host disease.

[0003] Gamma delta T cells (γδ T cells) represent a subset of T cells that express a distinct γδ T cell receptor (TCR) on their surface. This TCR is composed of one gamma gamma (γ) chain and one delta (δ) chain, each of which undergoes chain rearrangement but has a limited number of V genes compared to αβ T cells. The main TRGV gene segments encoding Vγ are TRGV2, TRGV3, TRGV4, TRGV5, TRGV8, TRGV9, and TRGV11, as well as nonfunctional genes TRGV10, TRGV11, TRGVA, and TRGVB. The most common TRGV gene segments encode Vδ1, Vδ2, and Vδ3, with some V segments designated as both Vδ and Vα (Adams et al., 2015, Cell Immunol., 296:30-40). Human γδ T cells can be broadly classified based on their TCR chain, as particular γ and δ types are more commonly, but not exclusively, found on cells in one or more tissue types. For example, most blood-resident γδ T cells express the Vδ2 TCR (commonly Vγ9Vδ2), whereas tissue-resident γδ T cells, such as those in the skin, more frequently use the Vδ1 TCR paired with a gamma chain, e.g., often paired with Vγ4 in the gut.

[0004] To utilize γδ T cells for immunotherapy, a means is required to either expand the cells in vivo or harvest and expand them ex vivo before reinfusion. The latter approach has previously been described using the addition of exogenous cytokines; see, for example, International Publication Nos. 2017 / 072367 and 2018 / 212808. Methods for expanding a patient's own γδ T cells have been described using pharmacologically modified forms of hydroxymethyl but-2-enyl pyrophosphate (HMBPP) or clinically approved aminobisphosphonates. These approaches appear to safely treat over 250 cancer patients, although the incidence of complete remission is rare. However, there remains a need for activating agents with a demonstrated ability to expand large numbers of γδ T cells.

[0005] Furthermore, binders or activators that can preferentially target, bind to, or recognize V51+ cells in a system, or specifically modulate or increase the number of V51+ cells, may be highly desirable as pharmaceuticals.

[0006] However, although medications exist that potentially modulate V52+ cells, including aminobisphosphonates such as Zometa® (zoledronic acid), these medications are primarily designed to slow bone resorption. Despite this V52+ modulation, there is a need to develop medications specifically designed to bind, target, modulate, activate, or increase the number of V51+ cells.

[0007] Furthermore, given the predominant tissue residency of V51+ cells, an ideal drug capable of modulating V51+ would also exhibit fewer "off-target" unwanted effects and rapid renal clearance. Typically, these unwanted effects can occur when using small molecule chemicals. For example, the aforementioned aminobisphosphonates, which have been shown to be able to modulate a distinct class of V52+ cells (secondary to their primary regulatory effect on bone), are associated with nephrotoxicity, manifested as worsening renal function and potential renal failure (e.g., Markowitz et al. (2003) Kidney Int. 64(1):281-289). Additional undesirable effects listed by the European Medicines Agency for Zometa include anemia, hypersensitivity reactions, hypertension, atrial fibrillation, muscle pain, generalized pain, fatigue, increased blood urea, vomiting, joint swelling, and chest pain.

[0008] Thus, there is a need for improved pharmaceuticals specifically designed to target V51+ cells and for the treatment of infectious diseases, autoimmune diseases, and cancer. Specifically, there is a need for pharmaceuticals that can be administered to alleviate the signs and symptoms of disease by specifically binding to, targeting, specifically activating, specifically enhancing the proliferation and / or cytotoxic activity of V51+ cells, or specifically blocking the activation of V51+ cells. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] International Publication No. 2017 / 072367 [Patent Document 2] International Publication No. 2018 / 212808 [Non-patent literature]

[0010] [Non-Patent Document 1] Adams et al.,296:30-40(2015)Cell Immunol. [Non-patent document 2] Markowitz et al. (2003) Kidney Int.64(1):281-289) Summary of the Invention

[0011] According to a first aspect, there is provided an anti-vδ1 antibody, or fragment thereof, for use in a method of treating cancer, an infectious disease, or an inflammatory disease. It will be understood that the methods and compositions for use described herein relate to administering the anti-vδ1 antibody, or fragment thereof, directly to the subject to be treated.

[0012] According to a further aspect of the invention there is provided an isolated multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first of the at least two target antigens is V51; CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26 to 37 and A1 to A12 (in Table 3), and / or Provided is an isolated multispecific antibody or fragment thereof, comprising one or more CDR1s comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 61.

[0013] According to a further aspect of the present invention there is provided an isolated human anti-TCR delta variable 1 multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first target antigen of the at least two target antigens is Vδ1, and the multispecific antibody or fragment thereof that binds to an epitope of Vδ1 comprises one or more amino acid residues within amino acids 1 to 90 of SEQ ID NO: 1.

[0014] According to a further aspect of the present invention there is provided an isolated multispecific antibody or fragment thereof as defined herein for use as a medicament.

[0015] According to a further aspect of the present invention there is provided an isolated multispecific antibody or fragment thereof as defined herein for use in the treatment of cancer, an infectious disease or an inflammatory disease. [Brief explanation of the drawings]

[0016] [Figure 1] ELISA detection of directly coated antigens using anti-V51 Ab (REA173, Miltenyi Biotec). Detection was only seen with antigens containing the V51 domain. The leucine zipper (LZ) format appeared to be more potent than the Fc format, consistent with cell-based flow competition assays (data not shown). [Figure 2] Polyclonal phage DELFIA data for DV1 selection. A) Heterodimer selection: heterodimeric LZ TCR format in rounds 1 and 2, with deselection for heterodimeric LZ TCR in both rounds. B) Homodimer selection: Round 1 was performed using homodimeric Fc fusion TCR with deselection for human IgG1 Fc, followed by round 2 with deselection for heterodimeric LZ TCR. Each graph contains two bars for each target, representing selections from different libraries. [Figure 3-1] IgG capture: Left) Sensorgram of the interaction of anti-L1 IgG with L1, Right) Steady-state fitting where available. All experiments were performed on a MASS-2 instrument at room temperature. Steady-state fitting with Langmuir 1:1 binding. [Figure 3-2] Same as above [Figure 3-3] Same as above [Figure 4]Results of TCR down-regulation assays for clones 1245_P01_E07, 1252_P01_C08, 1245_P02_G04, 1245_P01_B07 and 1251_P02_C05 (A), or clones 1139_P01_E04, 1245_P02_F07, 1245_P01_G06 1245_P01_G09, 1138_P01_B09, 1251_P02_G10 and 1252_P01_C08 (B). [Figure 5] Results of T cell degranulation assays for clones 1245_P01_E07, 1252_P01_C08, 1245_P02_G04, 1245_P01_B07, and 1251_P02_C05 (A), or clones 1139_P01_E04, 1245_P02_F07, 1245_P01_G06, 1245_P01_G09, 1138_P01_B09, and 1251_P02_G10 (B). [Figure 6] Results of killing assays (THP-1 flow-based assay) for clones 1245_P01_E07, 1252_P01_C08, 1245_P02_G04, 1245_P01_B07, and 1251_P02_C05 (A), or clones 1139_P01_E04, 1245_P02_F07, 1245_P01_G06, 1245_P01_G09, 1138_P01_B09, and 1251_P02_G10 (B). [Figure 7] Epitope mapping data for 1245_P01_E07. Graphical representation of the epitope binding site of 1245_P01_E07 on SEQ ID NO: 1. [Figure 8] Epitope mapping data for 1252_P01_C08. Graphical representation of the epitope binding site of 1252_P01_C08 on SEQ ID NO: 1. [Figure 9] Epitope mapping data for 1245_P02_G04. Graphical representation of the epitope binding site of 1245_P02_G04 on SEQ ID NO: 1. [Figure 10] Epitope mapping data for 1251_P02_C05. Graphical representation of the epitope binding site of 1251_P02_C05 on SEQ ID NO: 1. [Figure 11] Epitope mapping data for 1141_P01_E01. Graphical representation of the epitope binding site of 1141_P01_E01 on SEQ ID NO: 1. [Figure 12-1] Total cell counts in experiment 1 of Example 10. Samples were cultured with various concentrations of anti-V51 antibodies as described herein and compared to samples cultured with a comparator antibody or control. Graphs show total cell counts at (A) day 7, (B) day 14, and (C) day 18. [Figure 12-2] Same as above [Figure 12-3] Same as above [Figure 13-1] Analysis of V51 T cells in experiment 1 of Example 10. Graphs show (A) percentage of V51 T cells, (B) number of V51 T cells, (C) V51 fold change in day 18 samples. [Figure 13-2] Same as above [Figure 13-3] Same as above [Figure 14-1] Total cell counts in experiment 2 of Example 10. Samples were cultured with various concentrations of anti-V51 antibodies as described herein and compared to samples cultured with a comparator antibody or control. Graphs show total cell counts at (A) day 7, (B) day 11, (C) day 14, and (D) day 17. [Figure 14-2] Same as above [Figure 14-3] Same as above [Figure 14-4] Same as above [Figure 15-1] Analysis of V51 T cells in experiment 2 of Example 10. Graphs show (A) percentage of V51 T cells, (B) number of V51 T cells, (C) V51 fold change in day 17 samples. [Figure 15-2] Same as above [Figure 15-3] Same as above [Figure 16-1] Cell composition analysis. The cell types present in the samples (including non-V51 cells) were determined on day 17 of experiment 2. Cells were harvested and analyzed by flow cytometry for surface expression of V51, V52 and αβ TCR. Percentage values ​​are also provided in Table 6. [Figure 16-2]Same as above [Figure 17-1] SYTOX-Flow Killing Assay Results. Cell function was tested using the SYTOX-Flow Killing Assay, and results are presented for (A) Experiment 1 at day 14, using cells at an effector-to-target (E:T) ratio of 10:1, and (B) Experiment 2 at day 17 (post-freeze-thaw), using cells at E:T ratios of 1:1 and 10:1. [Figure 17-2] Same as above [Figure 18] Total cell counts after freeze-thaw. The graph shows the total cell counts after 7 days of freeze-thaw cultures for cultures that were contacted with B07, C08, E07, G04, or OKT-3 antibodies before freezing. [Figure 19] Monitoring cell expansion: Total cell number was monitored for cells cultured after freeze-thawing up to day 42. [Figure 20] Binding equivalence studies on engineered anti-Vd1 antibodies. [Figure 21] Anti-Vδ1 antibody binding equivalence studies of the human germline Vδ1 antigen and its polymorphic variants. [Figure 22] Anti-V51 antibodies resulted in increased V51+ cell cytokine secretion levels. Tissue-derived γδ T cells were incubated with antibodies as indicated. A) Observed levels of TNF-alpha, B) Observed levels of IFN-gamma. [Figure 23] Anti-Vδ1 antibodies resulted in increased Vδ1+ cell granzyme B levels / activity. Cancer cells were co-cultured with tissue-derived γδ T cells at a T:E ratio of 1:20 for 1 hour, along with the indicated antibodies. Results highlight the amount of granzyme B detected in cancer cells at the end of the co-culture. [Figure 24] Anti-V51 antibodies resulted in the modulation and proliferation of immune cells in human tissues. Human skin punch biopsies (from 5 different donors) were incubated for 21 days in culture with the indicated antibodies. A) Number of viable pan-γδ+ cells. B) Number of viable V51+ cells. C) Percentage of viable double-positive V51+CD25+ cells. [Figure 25-1]Anti-Vδ1 antibodies resulted in modulation and proliferation of tumor-infiltrating lymphocytes (TILs) in human tumors. Study with renal cell carcinoma (RCC) + / - antibodies. A) Fold increase in TIL Vδ1+ cells. B) Total number of TIL Vδ1+ cells. C) Example of gating strategy. D) Comparative cell surface phenotypic profile of TIL Vδ1+ cells. E) Analysis of TIL Vδ1-negative gated fraction. [Figure 25-2] Same as above [Figure 26-1] Anti-V51 antibodies resulted in enhanced V51+-mediated cytotoxicity and disease-cell-specific cytotoxicity. Cytotoxicity / potency assays in a model system involving triple cultures of V51+ effector cells, THP-1 monocytic cancer cells, and non-diseased healthy primary monocytes. A) Quantification of THP-1 and monocytic cell numbers in triple cocultures with γδ T cells in the presence of anti-V51 mAb or control. B) Bar graph representation highlighting the window between disease-cell-specific killing and non-diseased healthy cells: left bar graph, fold increase in killing of diseased cells (THP-1) relative to killing of non-diseased cells (primary human monocytes). right bar graph, same data but expressed as percent enhancement of killing relative to control. C) Tabulated results summarizing the percent improvement in efficacy of THP-1 target cells + / - mAbs in killing V51+ effector cells. D) Tabulated results of EC50 values ​​calculated from panel (A), expressed as the number of γδT cells required to cause 50% THP-1 cell killing. [Figure 26-2] Same as above [Figure 27-1]Multispecific antibodies resulted in enhanced V51+ effector cell-mediated cytotoxicity. Targeting tissue-centric disease-associated antigens: (A-D) Example of co-culture of V51+ effector cells with A-431 cancer cells + / - multispecific antibodies comprising an anti-V51 x anti-TAA(EGFr) bispecific binding moiety, in which the anti-V51 VL+VH binding domains (against the first target) are combined with the CH1-CH2-CH3 domains of the anti-EGFr binding moiety (against the second target). (E-H) Example of co-culture of V51+ effector cells with A-431 cancer cells + / - multispecific antibody comprising an anti-V51 x anti-TAA(EGFr) bispecific binding moiety, where the anti-V51 binding domain (against the first target) comprises a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), then combined with an anti-EGFr cetuximab-derived scFv binding moiety (against the second target). (I-J) Alternative approach to presenting the data: percentage improvement conferred by the multispecific antibody on the cytotoxicity of V51+ effector cells against EGFR+ cells compared to the component moieties. [Figure 27-2] Same as above [Figure 28] Multispecific antibodies resulted in enhanced V51+-mediated cytotoxicity and disease cell-specific cytotoxicity. Targeting hematopoietic disease-associated antigens. (A) E:T ratio required to induce 50% Raji cell killing, (B) improvement by addition of V51-CD19 multispecific antibodies. DETAILED DESCRIPTION OF THE INVENTION

[0017] definition Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the following terms have the meanings ascribed to them below.

[0018] Gamma delta (γδ) T cells represent a small subset of T cells that express a distinct T cell receptor (TCR) on their surface. This TCR is composed of one gamma (γ) chain and one delta (δ) chain. Each chain contains a variable (V) region, a constant (C) region, a transmembrane region, and a cytoplasmic tail. The V region contains the antigen-binding site. There are two major subtypes of human γδ T cells: those that predominate in peripheral blood and those that predominate in nonhematopoietic tissues. The two subtypes can be defined by the type of δ and / or γ present on the cell. For example, γδ T cells that predominate in peripheral blood primarily express the delta variable 2 chain (Vδ2). γδ T cells that predominate in nonhematopoietic tissues (i.e., tissue-resident) primarily express the delta variable 1 chain. Reference to "Vδ1 T cells" refers to γδ T cells that possess the Vδ1 chain, i.e., Vδ1+ cells.

[0019] References to "delta variable 1" may also be referred to as V51 or Vd1, while the nucleotides encoding the TCR chain comprising this region may be referred to as "TRDV1". Any antibody or fragment thereof that interacts with the V51 chain of a γδ TCR is an effective antibody or fragment thereof that binds to V51 and may be referred to as an "anti-TCR delta variable 1 antibody or fragment thereof" or an "anti-V51 antibody or fragment thereof".

[0020] Further reference is made herein to other delta chains, such as the "delta variable 2" chain. These may be referred to similarly. For example, the delta variable 2 chain may be referred to as V52, while the nucleotides encoding the TCR chain containing this region may be referred to as "TRDV2." In preferred embodiments, an antibody or fragment thereof that interacts with the V51 chain of a γδ TCR does not interact with other delta chains, such as V52.

[0021] Reference is also made herein to "gamma variable chains." These may also be referred to as gamma chains or Vγ, while the nucleotides encoding the TCR chain containing this region may be referred to as TRGV. For example, TRGV4 refers to the Vγ4 chain. In preferred embodiments, an antibody or fragment thereof that interacts with the Vδ1 chain of a γδ TCR does not interact with a gamma chain such as Vγ4.

[0022] The term "antibody" includes any antibody protein construct comprising at least one antibody variable domain containing at least one antigen-binding site (ABS). Antibodies include, but are not limited to, immunoglobulins of the IgA, IgG, IgE, IgD, and IgM types (and their subtypes). The overall structure of immunoglobulin G (IgG) antibodies, assembled from two identical heavy (H) chains and two identical light (L) chain polypeptides, is well established and highly conserved in mammals (Padlan (1994) Mol. Immunol. 31:169-217).

[0023] A conventional antibody or immunoglobulin (Ig) is a protein comprising four polypeptide chains: two heavy (H) chains and two light (L) chains. Each chain is divided into a constant region and a variable domain. The heavy (H) chain variable domain is abbreviated herein as VH, and the light (L) chain variable domain is abbreviated herein as VL. These domains, their related domains, and domains derived therefrom may be referred to herein as immunoglobulin chain variable domains. The VH and VL domains (also referred to as VH and VL regions) can be further subdivided into regions called "complementarity-determining regions" ("CDRs") and "framework regions" ("FRs"), interspersed with more conserved regions. The framework and complementarity-determining regions have been precisely defined (Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, (1991) NIH Publication Number 91-3242). Alternative numbering conventions for CDR sequences exist, such as those shown in Chothia et al. (1989) Nature 342:877-883. In conventional antibodies, each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A conventional antibody tetramer of two immunoglobulin heavy chains and two immunoglobulin light chains is formed, for example, by the immunoglobulin heavy and light chains interconnected by disulfide bonds and the heavy chains similarly connected. The heavy chain constant region contains three domains: CH1, CH2, and CH3. The light chain constant region is composed of one domain, CL. The heavy chain variable domain and the light chain variable domain are binding domains that interact with antigens. The constant regions of the antibodies typically mediate the binding of the antibody to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0024] As used herein, a fragment of an antibody (which may also be referred to as an "antibody fragment," "immunoglobulin fragment," "antigen-binding fragment," or "antigen-binding polypeptide") refers to a portion of an antibody (or a construct containing that portion) that specifically binds to a target, the delta variable 1 (V51) chain of the γδ T-cell receptor (e.g., a molecule in which one or more immunoglobulin chains are not full length, but which specifically binds to a target). Examples of binding fragments encompassed within the term antibody fragment include: (i) Fab fragment (a monovalent fragment consisting of the VL, VH, CL and CH1 domains); (ii) F(ab')2 fragment (a bivalent fragment consisting of two Fab fragments linked by a disulfide bridge at the hinge region); (iii) Fd fragment (consisting of the VH and CH1 domains); (iv) Fv fragments (consisting of the VL and VH domains of a single arm of an antibody); (v) single-chain variable fragments, scFv, which consist of VL and VH domains connected by a synthetic linker using recombinant methods, allowing them to be produced as a single protein chain in which the VL and VH regions pair to form monovalent molecules; (vi) VH (an immunoglobulin chain variable domain consisting of a VH domain), (vii) VL (an immunoglobulin chain variable domain consisting of a VL domain), (viii) domain antibodies (dAbs, consisting of either VH or VL domains); (ix) a minibody (consisting of a pair of scFv fragments linked via a CH3 domain); and (x) Diabodies, which consist of a non-covalent dimer of scFv fragments consisting of the VH domain from one antibody and the VL domain from another antibody connected by a small peptide linker.

[0025] "Human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. A human subject administered with such a human antibody does not generate an interspecies antibody response (e.g., a HAMA response—called human anti-mouse antibody) against primary amino acids contained within the antibody. Such a human antibody may contain amino acid residues (e.g., mutations introduced by random or site-specific mutagenesis or somatic mutation) not encoded by human germline immunoglobulin sequences, for example, in the CDRs, particularly CDR3. However, this term is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted onto human framework sequences. Human antibodies prepared, expressed, produced, or isolated by recombinant means, e.g., antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes, or antibodies prepared, expressed, produced, or isolated by any other means involving splicing of human immunoglobulin gene sequences into other DNA sequences, may also be referred to as "recombinant human antibodies."

[0026] The replacement of at least one amino acid residue in a framework region of a non-human immunoglobulin variable domain with the corresponding residue from a human variable domain is called "humanization." Humanization of variable domains can reduce immunogenicity in humans.

[0027] "Specificity" refers to the number of different types of antigens or antigenic determinants to which a particular antibody or fragment thereof can bind. Antibody specificity is the antibody's ability to recognize a particular antigen as a unique molecular entity and distinguish it from other antigens. An antibody that "specifically binds" to an antigen or epitope is a term well understood in the art. A molecule is said to exhibit "specific binding" if it reacts more frequently, more rapidly, with longer duration, and / or with higher affinity with a particular target antigen or epitope compared to alternative targets. An antibody "specifically binds" to a target antigen or epitope if it binds with higher affinity, avidity, more readily, and / or with longer duration than it binds to other substances.

[0028] "Affinity" is expressed by the equilibrium constant (KD) of dissociation between an antigen and an antigen-binding polypeptide, and is a measure of the binding strength between an antigenic determinant and an antigen-binding site on an antibody (or fragment thereof); the smaller the KD value, the stronger the binding strength between the antigenic determinant and the antigen-binding polypeptide. Alternatively, affinity can be expressed as an affinity constant (KA), which is 1 / KD. Affinity can be determined by known methods depending on the specific antigen of interest.

[0029] 10 -6 Any KD value less than is considered to indicate binding. Specific binding of an antibody or fragment thereof to an antigen or antigenic determinant can be determined by any suitable known method, including, for example, Scatchard analysis and / or competitive binding assays, such as radioimmunoassays (RIA), enzyme immunoassays (EIA) and sandwich competition assays, equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance, or spectroscopy (e.g., using a fluorescence assay), as well as different variants known in the art.

[0030] "Avidity" is a measure of the strength of binding between an antibody or fragment thereof and an associated antigen. Avidity is related to both the affinity between an antigenic determinant and its antigen-binding site on the antibody, as well as the number of associated binding sites present on the antibody.

[0031] "Human tissue V51+ cells," "hematopoietic and blood V51+ cells," and "tumor-infiltrating lymphocyte (TIL) V51+ cells" are defined as V51+ cells contained in or derived from either human tissues or the hematopoietic blood system or human tumors, respectively. All of the above cell types can be identified by their (i) location, or where they originate, and (ii) expression of the V51+ TCR.

[0032] A "modulatory antibody" is an antibody that, upon contact with or binding to a cell expressing a target to which the antibody binds, produces a measurable change, including, but not limited to, a measurable change in cell cycle, and / or cell number, and / or cell viability, and / or one or more cell surface markers, and / or secretion of one or more secreted molecules (e.g., cytokines, chemokines, leukotrienes, etc.), and / or function (e.g., cytotoxicity towards target cells or diseased cells). A method of "modulating" a cell or collection thereof refers to a method that triggers at least one measurable change in, or secretion from, the aforementioned cell or cells to produce one or more "modulated cells."

[0033] An "immune response" is a measurable change in at least one cell, or one cell type, or one endocrine pathway, or one exocrine pathway of the immune system (including, but not limited to, cell-mediated response, humoral response, cytokine response, chemokine response) upon addition of a regulatory antibody.

[0034] "Immune cells" are defined as cells of the immune system, including, but not limited to, CD34+ cells, B cells, CD45+ (lymphocyte common antigen) cells, alpha-beta T cells, cytotoxic T cells, helper T cells, plasma cells, neutrophils, monocytes, macrophages, red blood cells, platelets, dendritic cells, phagocytes, granulocytes, innate lymphoid cells, natural killer (NK) cells, and gamma delta T cells. Typically, immune cells are classified using combinatorial cell surface molecular analysis (e.g., via flow cytometry) to identify, group, or cluster immune cells for differentiation into subpopulations. These can then be further subdivided with additional analysis. For example, CD45+ lymphocytes can be further subdivided into vδ-positive and vδ-negative populations.

[0035] A "model system" is a biological model or representation designed to aid in understanding how a pharmaceutical agent, such as an antibody or fragment thereof, may function as a pharmaceutical agent in alleviating the signs or symptoms of a disease. Such models typically include the use of diseased cells, non-diseased cells, healthy effector cells, and tissues in vitro, ex vivo, and in vivo to study and compare the performance of such pharmaceutical agents.

[0036] A "disease cell" exhibits a phenotype associated with a disease such as cancer, an infection such as a viral infection, or the progression of an inflammatory condition or disease. For example, the disease cell may be a tumor cell, an autoimmune tissue cell, or a virally infected cell. Thus, the disease cell may be defined as being neoplastic, virally infected, or inflammatory.

[0037] "Healthy cells" refer to normal cells that are not diseased. They can also be referred to as "normal" or "non-disease" cells. Non-disease cells include non-cancerous, non-infectious, or non-inflammatory cells. These cells are often used together with related diseased cells to determine the diseased cell specificity of a drug and / or to better understand the therapeutic index of a drug.

[0038] "Disease cell specificity" is a measure of how effectively an effector cell or a population thereof (e.g., a population of V51+ cells) can distinguish and kill diseased cells, such as cancer cells, while sparing non-disease or healthy cells. This potential can be measured in a model system and can involve comparing the tendency of an effector cell or a population of effector cells to selectively kill or lyse diseased cells with the potential of said effector cells to kill or lyse non-disease or healthy cells. Such disease cell specificity can inform the potential therapeutic index of a pharmaceutical.

[0039] "Enhanced disease cell specificity" describes the phenotype of an effector cell, e.g., a V51+ cell, or a population thereof, that has been modulated to further increase its ability to specifically kill disease cells. This enhancement can be measured in a variety of ways, including the fold change or percentage increase in disease cell killing specificity or selectivity.

[0040] Preferably, the antibody or fragment thereof (i.e., polypeptide) is isolated. An "isolated" polypeptide is a polypeptide that is removed from its original environment. The term "isolated" can be used to refer to an antibody that is substantially free of other antibodies with different antigen specificities (e.g., an isolated antibody that specifically binds to Vδ1, or a fragment thereof, is substantially free of antibodies that specifically bind to antigens other than Vδ1). The term "isolated" can also be used to refer to a preparation in which the isolated antibody is sufficiently pure to be administered therapeutically when formulated as the active ingredient of a pharmaceutical composition, or is at least 70-80% (w / w) pure, more preferably at least 80-90% (w / w) pure, even more preferably 90-95% pure, or most preferably at least 95%, 96%, 97%, 98%, 99%, or 100% (w / w) pure.

[0041] Preferably, the polynucleotide used in the present invention is isolated.An "isolated" polynucleotide is a polynucleotide that is removed from its original environment.For example, a naturally occurring polynucleotide is isolated when it is separated from some or all of the materials that coexist in natural system.For example, if a polynucleotide is cloned into a vector that is not part of its natural environment, or if the polynucleotide is contained in cDNA, it is considered to be isolated.

[0042] An antibody or fragment thereof may be a "functionally active variant," including naturally occurring allelic variants as well as mutants or any non-naturally occurring variants. As known in the art, allelic variants are alternative forms of (poly)peptides characterized by one or more amino acid substitutions, deletions, or additions that do not essentially alter the biological function of the polypeptide. As a non-limiting example, the above-mentioned functionally active variants may still function when the framework containing the CDRs is modified, when the CDRs themselves are modified, when the above-mentioned CDRs are grafted onto alternative frameworks, or when N- or C-terminal extensions are incorporated. Furthermore, the CDRs containing the binding domains may pair with different partner chains, such as those shared with another antibody. When shared with a so-called "common" light chain or a "common" heavy chain, the above-mentioned binding domains may still function. Furthermore, the above-mentioned binding domains may function when multimerized. Additionally, an "antibody or fragment thereof" may include functional variants in which the VH or VL or constant domains have been altered away from or towards different canonical sequences (e.g., as listed on IMGT.org) and still function.

[0043] For purposes of comparing two closely related polypeptide sequences, the "% sequence identity" between a first polypeptide sequence and a second polypeptide sequence may be calculated using NCBI BLAST v2.0 using standard settings for polypeptide sequences (BLASTP). For purposes of comparing two closely related polynucleotide sequences, the "% sequence identity" between a first nucleotide sequence and a second nucleotide sequence may be calculated using NCBI BLAST v2.0 using standard settings for nucleotide sequences (BLASTN).

[0044] A polypeptide or polynucleotide sequence is said to be the same as, or "identical to," another polypeptide or polynucleotide sequence if the sequences share 100% sequence identity over their entire length. Residues in a sequence are numbered from left to right, i.e., from N-terminus to C-terminus for polypeptides and from 5' to 3' for polynucleotides.

[0045] A "difference" between sequences refers to the insertion, deletion, or substitution of a single amino acid residue at a position in the second sequence compared to the first sequence. Two polypeptide sequences can contain one, two, or more such amino acid differences. An insertion, deletion, or substitution in a second sequence that is otherwise identical (100% sequence identity) to the first sequence reduces the percent sequence identity. For example, if an identical sequence is 9 amino acid residues long, a single substitution in the second sequence results in 88.9% sequence identity. If a first and second polypeptide sequence are 9 amino acid residues long and share 6 identical residues, the first and second polypeptide sequences share more than 66% identity (the first and second polypeptide sequences share 66.7% identity).

[0046] Alternatively, for the purpose of comparing a first reference polypeptide sequence with a second comparison polypeptide sequence, the number of additions, substitutions, and / or deletions made to the first sequence to generate the second sequence may be determined. An "addition" is the addition of one amino acid residue to the sequence of the first polypeptide (including additions at either end of the first polypeptide). A "substitution" is the replacement of one amino acid residue in the sequence of the first polypeptide with a different amino acid residue. This substitution may be conservative or non-conservative. A "deletion" is the deletion of one amino acid residue from the sequence of the first polypeptide (including deletions at either end of the first polypeptide).

[0047] A "conservative" amino acid substitution is one in which an amino acid residue is replaced with another amino acid residue having a similar chemical structure and is expected to have little effect on the function, activity, or other biological properties of a polypeptide. Preferably, such a conservative substitution is one in which one amino acid residue within the following group is replaced with another amino acid residue within the same group: JPEG2025160218000001.jpg139166

[0048] Preferably, the hydrophobic amino acid residue is a non-polar amino acid. More preferably, the hydrophobic amino acid residue is selected from V, I, L, M, F, W, or C.

[0049] As used herein, the numbering of polypeptide sequences and the definitions of CDR and FR are as defined according to the Kabat system (Kabat et al., 1991, incorporated herein by reference in its entirety).The "corresponding" amino acid residue between a first polypeptide sequence and a second polypeptide sequence is the amino acid residue in the first sequence that shares the same position according to the Kabat system with the amino acid residue in the second sequence, but the amino acid residue in the second sequence may have different identity with the first sequence.Appropriately, corresponding residues share the same number (and letter) when the framework and CDR are the same length according to Kabat definition.Alignment can be achieved manually or by using known computer algorithms for sequence alignment, such as, for example, NCBI BLAST v2.0 (BLASTP or BLASTN) using standard settings.

[0050] The term "epitope" herein refers to the portion of a target that is specifically bound by an antibody or its fragment. An epitope can also be referred to as an "antigenic determinant." An antibody binds to "essentially the same epitope" as another antibody when both recognize the same or sterically overlapping epitopes. A commonly used method for determining whether two antibodies bind to the same or overlapping epitopes is a competitive assay, which can be configured in several different formats (e.g., well plates using radioactive or enzyme labels, or flow cytometry on antigen-expressing cells) using either labeled antigen or labeled antibody.

[0051] Epitopes found on protein targets can be defined as "linear epitopes" or "conformational epitopes." Linear epitopes are formed by contiguous sequences of amino acids in the protein antigen. Conformational epitopes are formed from amino acids that are discontinuous in the protein sequence but come together when the protein folds into its three-dimensional structure.

[0052] The term "vector," as used herein, is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, into which additional DNA segments can be ligated. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian and yeast vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, thereby replicating along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. As used herein, "plasmid" and "vector" can be used interchangeably, as the plasmid is the most commonly used form of vector. However, the invention is intended to include other forms of expression vectors, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses and adeno-associated viruses), as well as bacteriophage and phagemid systems, which serve equivalent functions. The term "recombinant host cell" (or simply "host cell"), as used herein, is intended to refer to a cell into which a recombinant expression vector has been introduced. Such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell, for example, where such progeny are used to generate cell lines or cell banks, which are then optionally stored, provided, sold, transferred, or used to produce antibodies or fragments thereof as described herein.

[0053] References to a "subject," "patient," or "individual" refer to a subject to be treated, particularly a mammalian subject. Mammalian subjects include humans, non-human primates, farm animals (such as cows), sport animals, or pet animals such as dogs, cats, guinea pigs, rabbits, rats, or mice. In some embodiments, the subject is a human. In alternative embodiments, the subject is a non-human mammal, such as a mouse.

[0054] The term "sufficient amount" means an amount sufficient to produce a desired effect. The term "therapeutically effective amount" is an amount effective to alleviate the symptoms of a disease or disorder. Since prevention can be considered therapy, a therapeutically effective amount can be a "prophylactically effective amount."

[0055] A disease or disorder is "alleviated" if the severity of a sign or symptom of the disease or disorder, the frequency with which such sign or symptom is experienced by a subject, or both, is reduced.

[0056] As used herein, "treating a disease or disorder" means reducing the frequency and / or severity of at least one sign or symptom of the disease or disorder experienced by a subject.

[0057] "Cancer," as used herein, refers to the abnormal growth or division of cells. Generally, the growth and / or lifespan of cancer cells exceeds and is uncoordinated with the growth and / or lifespan of normal cells and surrounding tissues. Cancers can be benign, premalignant, or malignant. Cancer arises in a variety of cells and tissues, including the oral cavity (e.g., mouth, tongue, pharynx, etc.), digestive system (e.g., esophagus, stomach, small intestine, colon, rectum, liver, bile duct, gallbladder, pancreas, etc.), respiratory system (e.g., larynx, lungs, bronchi, etc.), bones, joints, skin (e.g., basal cell, squamous, meningioma, etc.), breast, reproductive system (e.g., uterus, ovaries, prostate, testes, etc.), urinary system (e.g., bladder, kidneys, ureters, etc.), eyes, nervous system (e.g., brain, etc.), endocrine system (e.g., thyroid, etc.), and hematopoietic system (e.g., lymphoma, myeloma, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, etc.).

[0058] As used herein, the term "about" as used herein means up to 10% greater than, and including, the specified value, up to 10% less than, and including, the specified value, preferably up to 5% greater than, and including, the specified value, up to 5% less than, and including, the specified value, and particularly including, the specified value. The term "between" includes the boundary values ​​specified.

[0059] Antibodies or fragments thereof Provided herein is an antibody or fragment thereof capable of specifically binding to the delta variable 1 chain (V51) of the gamma delta T cell receptor (TCR). The invention relates to the use of the above-mentioned antibody as a medicament for administration to a subject to be treated.

[0060] In one embodiment, the antibody or fragment thereof is an scFv, Fab, Fab', F(ab'), Fv, a variable domain (e.g., VH or VL), a diabody, a minibody, or a monoclonal antibody. In a further embodiment, the antibody or fragment thereof is an scFv.

[0061] The antibodies of the present invention may be of any class, e.g., IgG, IgA, IgM, IgE, IgD, or their isotypes, and may contain a kappa or lambda light chain. In one embodiment, the antibody is an IgG antibody, e.g., at least one of the isotypes IgG1, IgG2, IgG3, or IgG4. In a further embodiment, the antibody may be in a modified format (e.g., an IgG format) to confer desired properties, such as an Fc mutated to reduce effector function, extend half-life, alter ADCC, or improve hinge stability. Such modifications are well known in the art.

[0062] In one embodiment, the antibody or fragment thereof is human. Thus, the antibody or fragment thereof may be derived from a human immunoglobulin (Ig) sequence. The CDR, framework and / or constant region of the antibody (or fragment thereof) may be derived from a human Ig sequence, particularly a human IgG sequence. The CDR, framework and / or constant region may be substantially identical to a human Ig sequence, particularly a human IgG sequence. The advantage of using a human antibody is that human antibodies are less immunogenic or non-immunogenic in humans.

[0063] The antibody or fragment thereof may also be chimeric, for example, a mouse-human antibody chimera.

[0064] Alternatively, the antibody or fragment thereof is derived from a non-human species, such as a mouse. Such non-human antibodies can be modified to increase their similarity to antibody variants naturally produced in humans, such that the antibody or fragment thereof can be partially or fully humanized. Thus, in one embodiment, the antibody or fragment thereof is humanized.

[0065] Antibody sequence The isolated anti-V51 antibodies or fragments thereof of the invention may be described with reference to their CDR sequences.

[0066] According to one aspect of the invention there is provided an isolated anti-V51 antibody or fragment thereof, comprising: CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26 to 37 and A1 to A12, and / or Provided is an isolated anti-Vδ1 antibody or a fragment thereof, comprising one or more CDR1s comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 61.

[0067] According to one aspect of the present invention, there is provided an isolated anti-Vδ1 antibody or fragment thereof, comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 25. In one embodiment, the antibody or fragment thereof comprises a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26 to 37 and sequences A1 to A12 (in Table 3). In one embodiment, the antibody or fragment thereof comprises a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 61.

[0068] In one embodiment, the antibody or fragment thereof comprises a CDR3 comprising a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 2-25. In one embodiment, the antibody or fragment thereof comprises a CDR2 comprising a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 26-37 and sequences A1-A12 (of Table 3). In one embodiment, the antibody or fragment thereof comprises a CDR1 comprising a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 38-61.

[0069] In one embodiment, the antibody or fragment thereof comprises a CDR3 consisting of a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 2-25. In one embodiment, the antibody or fragment thereof comprises a CDR2 consisting of a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 26-37 and sequences A1-A12 (in Table 3). In one embodiment, the antibody or fragment thereof comprises a CDR1 consisting of a sequence having at least 85%, 90%, 95%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 38-61.

[0070] According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25. According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25.

[0071] According to a specific aspect of the present invention, there is provided an antibody or a fragment thereof, comprising: a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, or 4; and / or a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 15, or 16. According to another aspect of the present invention, there is provided an antibody or a fragment thereof, comprising: a VH region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, or 4; and / or a VL region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 15, or 16.

[0072] According to a specific aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 8 to 13, particularly 8, 9, 10, or 11, and / or a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 20 to 25, particularly 20, 21, 22, or 23. According to another aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 8 to 13, particularly 8, 9, 10, or 11, and / or a VL region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 20 to 25, particularly 20, 21, 22, or 23.

[0073] According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 14 to 25. According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 14 to 25.

[0074] According to a specific aspect of the present invention, there is provided an antibody or a fragment thereof comprising a VH region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, 4, or 5, and / or a VL region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 14, 15, or 17. According to another aspect of the present invention, there is provided an antibody or a fragment thereof comprising a VH region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, 4, or 5, and / or a VL region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 15, 16, or 17.

[0075] According to a particular aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 8, 9, 10, or 11, and / or a VL region comprising a CDR3 comprising a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 20, 21, 22, or 23. According to another aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 8, 9, 10, or 11, and / or a VL region comprising a CDR3 consisting of a sequence having at least 90% sequence identity to any one of SEQ ID NOs: 20, 21, 22, or 23.

[0076] According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 14 to 25. According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 2 to 13, and / or a VL region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 14 to 25.

[0077] According to a specific aspect of the present invention, there is provided an antibody or a fragment thereof comprising a VH region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, 4, or 5, and / or a VL region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 15, 16, or 17. According to another aspect of the present invention, there is provided an antibody or a fragment thereof comprising a VH region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 2 to 7, particularly 2 to 6, for example, 2, 3, 4, or 5, and / or a VL region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 14 to 19, particularly 14 to 18, for example, 14, 15, 16, or 17.

[0078] According to a particular aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 8, 9, 10, or 11, and / or a VL region comprising a CDR3 comprising a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 20, 21, 22, or 23. According to another aspect of the present invention, there is provided an antibody or a fragment thereof, comprising a VH region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 8, 9, 10, or 11, and / or a VL region comprising a CDR3 consisting of a sequence having at least 95% sequence identity to any one of SEQ ID NOs: 20, 21, 22, or 23.

[0079] According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising: a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13; and a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25. According to a further aspect of the present invention, there is provided an antibody or a fragment thereof, comprising: a VH region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13; and a VL region comprising a CDR3 consisting of a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25.

[0080] Embodiments referred to herein as "at least 80%" or "80% or greater" will be understood to include all values ​​of 80% or greater, e.g., 85%, 90%, 95%, 97%, 98%, 99%, or 100% sequence identity, etc. In one embodiment, an antibody or fragment of the invention comprises at least 85%, e.g., at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the designated sequence.

[0081] Instead of percent sequence identity, embodiments may be defined using one or more amino acid changes, for example, one or more additions, substitutions, and / or deletions. In one embodiment, the sequence may contain up to five amino acid changes, for example, up to three amino acid changes, particularly up to two amino acid changes. In a further embodiment, the sequence may contain up to five amino acid substitutions, for example, up to three amino acid substitutions, particularly up to one or two amino acid substitutions. For example, the CDR3 of the antibody or fragment thereof of the present invention comprises, or more preferably consists of, a sequence having two or fewer substitutions, more preferably one or fewer substitutions, compared to any one of SEQ ID NOs: 2 to 25.

[0082] Suitably, residues in any of CDR1, CDR2 or CDR3 that differ from their corresponding residues in SEQ ID NOs: 2-61 and A1-A12 are conservative substitutions with respect to their corresponding residues, for example, any residue in CDR3 that differs from its corresponding residue in SEQ ID NOs: 2-25 is a conservative substitution with respect to its corresponding residue.

[0083] In one embodiment, the antibody or fragment thereof (i) a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13; (ii) a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26 to 37; (iii) a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 49; (iv) a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25; (v) a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1 to A12, and / or (vi) A VL region including a CDR1 having a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 50 to 61.

[0084] In one embodiment, the antibody or fragment thereof (i) a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2 to 13; (ii) a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26 to 37; and (iii) A heavy chain having a VH region including a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 49.

[0085] In one embodiment, the antibody or fragment thereof (i) a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14 to 25; (ii) a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1 to A12; and (iii) A light chain having a VL region including a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 50 to 61.

[0086] In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2, 3, 4, 5, or 6, such as 2, 3, 4, or 5, particularly 2, 3, or 4. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26, 27, 28, 29, or 30, such as 26, 27, 28, or 29, particularly 26, 27, or 28. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38, 39, 40, 41, or 42, such as 38, 39, 40, or 41, particularly 38, 39, or 40.

[0087] In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 8, 9, 10, or 11. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 32, 33, 34, or 35. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 44, 45, 46, or 47.

[0088] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 2, a CDR2 comprising the sequence of SEQ ID NO: 26, and a CDR1 comprising the sequence of SEQ ID NO: 38. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 2, CDR2 consists of the sequence of SEQ ID NO: 26, and CDR1 consists of the sequence of SEQ ID NO: 38.

[0089] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 3, a CDR2 comprising the sequence of SEQ ID NO: 27, and a CDR1 comprising the sequence of SEQ ID NO: 39. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 3, CDR2 consists of the sequence of SEQ ID NO: 27, and CDR1 consists of the sequence of SEQ ID NO: 39.

[0090] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 4, a CDR2 comprising the sequence of SEQ ID NO: 28, and a CDR1 comprising the sequence of SEQ ID NO: 40. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 4, CDR2 consists of the sequence of SEQ ID NO: 28, and CDR1 consists of the sequence of SEQ ID NO: 40.

[0091] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 5, a CDR2 comprising the sequence of SEQ ID NO: 29, and a CDR1 comprising the sequence of SEQ ID NO: 41. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 5, CDR2 consists of the sequence of SEQ ID NO: 29, and CDR1 consists of the sequence of SEQ ID NO: 41.

[0092] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 6, a CDR2 comprising the sequence of SEQ ID NO: 30, and a CDR1 comprising the sequence of SEQ ID NO: 42. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 6, CDR2 consists of the sequence of SEQ ID NO: 30, and CDR1 consists of the sequence of SEQ ID NO: 42.

[0093] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 8, a CDR2 comprising the sequence of SEQ ID NO: 32, and a CDR1 comprising the sequence of SEQ ID NO: 44. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 8, CDR2 consists of the sequence of SEQ ID NO: 32, and CDR1 consists of the sequence of SEQ ID NO: 44.

[0094] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 9, a CDR2 comprising the sequence of SEQ ID NO: 33, and a CDR1 comprising the sequence of SEQ ID NO: 45. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 9, CDR2 consists of the sequence of SEQ ID NO: 33, and CDR1 consists of the sequence of SEQ ID NO: 45.

[0095] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 10, a CDR2 sequence of SEQ ID NO: 34, and a CDR1 sequence of SEQ ID NO: 46. In one embodiment, the CDR3 consists of the sequence of SEQ ID NO: 10, the CDR2 consists of the sequence of SEQ ID NO: 34, and the CDR1 consists of the sequence of SEQ ID NO: 46.

[0096] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 11, a CDR2 sequence of SEQ ID NO: 35, and a CDR1 sequence of SEQ ID NO: 47. In one embodiment, the CDR3 consists of the sequence of SEQ ID NO: 11, the CDR2 consists of the sequence of SEQ ID NO: 35, and the CDR1 consists of the sequence of SEQ ID NO: 47.

[0097] In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14-25, e.g., SEQ ID NOs: 14, 15, 16, 17, or 18, e.g., 14, 15, 16, or 17, particularly 14, 15, or 16. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1-A12 (of Table 3), e.g., sequence A1, A2, A3, A4, or A5, e.g., A1, A2, A3, or A4, particularly A1, A2, or A3. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 50 to 61, e.g., SEQ ID NOs: 50, 51, 52, 53, or 54, e.g., 50, 51, 52, or 53, particularly 50, 51, or 52.

[0098] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 14, a CDR2 comprising the sequence of sequence A1, and a CDR1 comprising the sequence of SEQ ID NO: 50. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 14, CDR2 consists of the sequence of sequence A1, and CDR1 consists of the sequence of SEQ ID NO: 50.

[0099] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 15, a CDR2 comprising the sequence of sequence A2, and a CDR1 comprising the sequence of SEQ ID NO: 51. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 15, CDR2 consists of the sequence of sequence A2, and CDR1 consists of the sequence of SEQ ID NO: 51.

[0100] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 16, a CDR2 comprising the sequence of sequence A3, and a CDR1 comprising the sequence of SEQ ID NO: 52. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 16, CDR2 consists of the sequence of sequence A3, and CDR1 consists of the sequence of SEQ ID NO: 52.

[0101] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 17, a CDR2 comprising the sequence of sequence A4, and a CDR1 comprising the sequence of SEQ ID NO: 53. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 17, CDR2 consists of the sequence of sequence A4, and CDR1 consists of the sequence of SEQ ID NO: 53.

[0102] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 18, a CDR2 comprising the sequence of sequence A5, and a CDR1 comprising the sequence of SEQ ID NO: 54. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 18, CDR2 consists of the sequence of sequence A5, and CDR1 consists of the sequence of SEQ ID NO: 54.

[0103] In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 20, 21, 22, or 23. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: A7, A8, A9, or A10. In one embodiment, the antibody or fragment thereof comprises (or consists of) a VL region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 56, 57, 58, or 59.

[0104] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 20, a CDR2 comprising the sequence of sequence A7, and a CDR1 comprising the sequence of SEQ ID NO: 56. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 20, CDR2 consists of the sequence of sequence A7, and CDR1 consists of the sequence of SEQ ID NO: 56.

[0105] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 21, a CDR2 comprising the sequence of sequence A8, and a CDR1 comprising the sequence of SEQ ID NO: 57. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 21, CDR2 consists of the sequence of sequence A8, and CDR1 consists of the sequence of SEQ ID NO: 57.

[0106] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 22, a CDR2 comprising the sequence of sequence A9, and a CDR1 comprising the sequence of SEQ ID NO: 58. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 22, CDR2 consists of the sequence of sequence A9, and CDR1 consists of the sequence of SEQ ID NO: 58.

[0107] In one embodiment, the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 23, a CDR2 comprising the sequence of sequence A10, and a CDR1 comprising the sequence of SEQ ID NO: 59. In one embodiment, CDR3 consists of the sequence of SEQ ID NO: 23, CDR2 consists of the sequence of sequence A10, and CDR1 consists of the sequence of SEQ ID NO: 59.

[0108] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 2, a CDR2 comprising the sequence of SEQ ID NO: 26, and a CDR1 comprising the sequence of SEQ ID NO: 38, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 14, a CDR2 comprising the sequence of sequence A1, and a CDR1 comprising the sequence of SEQ ID NO: 50. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 2, the HCDR2 consists of the sequence of SEQ ID NO: 26, the HCDR1 consists of the sequence of SEQ ID NO: 38, the LCDR3 consists of the sequence of SEQ ID NO: 14, the LCDR2 consists of the sequence of sequence A1, and the LCDR1 consists of the sequence of SEQ ID NO: 50.

[0109] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 3, a CDR2 comprising the sequence of SEQ ID NO: 27, and a CDR1 comprising the sequence of SEQ ID NO: 39, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 15, a CDR2 comprising the sequence of sequence A2, and a CDR1 comprising the sequence of SEQ ID NO: 51. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 3, the HCDR2 consists of the sequence of SEQ ID NO: 27, the HCDR1 consists of the sequence of SEQ ID NO: 39, the LCDR3 consists of the sequence of SEQ ID NO: 15, the LCDR2 consists of the sequence of sequence A2, and the LCDR1 consists of the sequence of SEQ ID NO: 51.

[0110] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 4, a CDR2 comprising the sequence of SEQ ID NO: 28, and a CDR1 comprising the sequence of SEQ ID NO: 40, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 16, a CDR2 comprising the sequence of sequence A3, and a CDR1 comprising the sequence of SEQ ID NO: 52. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 4, the HCDR2 consists of the sequence of SEQ ID NO: 28, the HCDR1 consists of the sequence of SEQ ID NO: 40, the LCDR3 consists of the sequence of SEQ ID NO: 16, the LCDR2 consists of the sequence of sequence A3, and the LCDR1 consists of the sequence of SEQ ID NO: 52.

[0111] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 5, a CDR2 comprising the sequence of SEQ ID NO: 29, and a CDR1 comprising the sequence of SEQ ID NO: 41, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 17, a CDR2 comprising the sequence of sequence A4, and a CDR1 comprising the sequence of SEQ ID NO: 53. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 5, the HCDR2 consists of the sequence of SEQ ID NO: 29, the HCDR1 consists of the sequence of SEQ ID NO: 41, the LCDR3 consists of the sequence of SEQ ID NO: 17, the LCDR2 consists of the sequence of sequence A4, and the LCDR1 consists of the sequence of SEQ ID NO: 53.

[0112] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 6, a CDR2 comprising the sequence of SEQ ID NO: 30, and a CDR1 comprising the sequence of SEQ ID NO: 42, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 18, a CDR2 comprising the sequence of sequence A5, and a CDR1 comprising the sequence of SEQ ID NO: 54. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 6, the HCDR2 consists of the sequence of SEQ ID NO: 30, the HCDR1 consists of the sequence of SEQ ID NO: 42, the LCDR3 consists of the sequence of SEQ ID NO: 18, the LCDR2 consists of the sequence of sequence A5, and the LCDR1 consists of the sequence of SEQ ID NO: 54.

[0113] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 7, a CDR2 comprising the sequence of SEQ ID NO: 31, and a CDR1 comprising the sequence of SEQ ID NO: 43, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 19, a CDR2 comprising the sequence of sequence A6, and a CDR1 comprising the sequence of SEQ ID NO: 55. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 7, the HCDR2 consists of the sequence of SEQ ID NO: 31, the HCDR1 consists of the sequence of SEQ ID NO: 43, the LCDR3 consists of the sequence of SEQ ID NO: 19, the LCDR2 consists of the sequence of sequence A6, and the LCDR1 consists of the sequence of SEQ ID NO: 55.

[0114] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 8, a CDR2 comprising the sequence of SEQ ID NO: 32, and a CDR1 comprising the sequence of SEQ ID NO: 44, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 20, a CDR2 comprising the sequence of sequence A7, and a CDR1 comprising the sequence of SEQ ID NO: 56. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 8, the HCDR2 consists of the sequence of SEQ ID NO: 32, the HCDR1 consists of the sequence of SEQ ID NO: 44, the LCDR3 consists of the sequence of SEQ ID NO: 20, the LCDR2 consists of the sequence of sequence A7, and the LCDR1 consists of the sequence of SEQ ID NO: 56.

[0115] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 9, a CDR2 comprising the sequence of SEQ ID NO: 33, and a CDR1 comprising the sequence of SEQ ID NO: 45, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 21, a CDR2 comprising the sequence of sequence A8, and a CDR1 comprising the sequence of SEQ ID NO: 57. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 9, the HCDR2 consists of the sequence of SEQ ID NO: 33, the HCDR1 consists of the sequence of SEQ ID NO: 45, the LCDR3 consists of the sequence of SEQ ID NO: 21, the LCDR2 consists of the sequence of sequence A8, and the LCDR1 consists of the sequence of SEQ ID NO: 57.

[0116] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 10, a CDR2 comprising the sequence of SEQ ID NO: 34, and a CDR1 comprising the sequence of SEQ ID NO: 46, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 22, a CDR2 comprising the sequence of sequence A9, and a CDR1 comprising the sequence of SEQ ID NO: 58. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 10, the HCDR2 consists of the sequence of SEQ ID NO: 34, the HCDR1 consists of the sequence of SEQ ID NO: 46, the LCDR3 consists of the sequence of SEQ ID NO: 22, the LCDR2 consists of the sequence of sequence A9, and the LCDR1 consists of the sequence of SEQ ID NO: 58.

[0117] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 11, a CDR2 comprising the sequence of SEQ ID NO: 35, and a CDR1 comprising the sequence of SEQ ID NO: 47, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 23, a CDR2 comprising the sequence of sequence A10, and a CDR1 comprising the sequence of SEQ ID NO: 59. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 11, the HCDR2 consists of the sequence of SEQ ID NO: 35, the HCDR1 consists of the sequence of SEQ ID NO: 47, the LCDR3 consists of the sequence of SEQ ID NO: 23, the LCDR2 consists of the sequence of sequence A10, and the LCDR1 consists of the sequence of SEQ ID NO: 59.

[0118] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 12, a CDR2 comprising the sequence of SEQ ID NO: 36, and a CDR1 comprising the sequence of SEQ ID NO: 48, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 24, a CDR2 comprising the sequence of sequence A11, and a CDR1 comprising the sequence of SEQ ID NO: 60. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 12, the HCDR2 consists of the sequence of SEQ ID NO: 36, the HCDR1 consists of the sequence of SEQ ID NO: 48, the LCDR3 consists of the sequence of SEQ ID NO: 24, the LCDR2 consists of the sequence of sequence A11, and the LCDR1 consists of the sequence of SEQ ID NO: 60.

[0119] In one embodiment, the VH region comprises a CDR3 comprising the sequence of SEQ ID NO: 13, a CDR2 comprising the sequence of SEQ ID NO: 37, and a CDR1 comprising the sequence of SEQ ID NO: 49, and the VL region comprises a CDR3 comprising the sequence of SEQ ID NO: 25, a CDR2 comprising the sequence of sequence A12, and a CDR1 comprising the sequence of SEQ ID NO: 61. In one embodiment, the HCDR3 consists of the sequence of SEQ ID NO: 13, the HCDR2 consists of the sequence of SEQ ID NO: 37, the HCDR1 consists of the sequence of SEQ ID NO: 49, the LCDR3 consists of the sequence of SEQ ID NO: 25, the LCDR2 consists of the sequence of sequence A12, and the LCDR1 consists of the sequence of SEQ ID NO: 61.

[0120] In one embodiment, the antibody or fragment thereof comprises one or more CDR sequences listed in Table 3. In a further embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1252_P01_C08 listed in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P01_E07 listed in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P02_G04 listed in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P02_B07 listed in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1251_P02_C05 listed in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1139_P01_E04 described in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P02_F07 described in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P01_G06 described in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1245_P01_G09 described in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1138_P01_B09 described in Table 3. In an alternative embodiment, the antibody or fragment thereof comprises one or more (e.g., all) CDR sequences of clone 1251_P02_G10 described in Table 3.

[0121] Preferably, the VH and VL regions listed above each comprise four framework regions (FR1-FR4). In one embodiment, the antibody or fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and / or FR4) comprising a sequence having at least 80% sequence identity to a framework region in any one of SEQ ID NOs: 62-85. In one embodiment, the antibody or fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and / or FR4) comprising a sequence having at least 90%, e.g., at least 95%, 97%, or 99% sequence identity to a framework region in any one of SEQ ID NOs: 62-85. In one embodiment, the antibody or fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and / or FR4) comprising a sequence in any one of SEQ ID NOs: 62-85. In one embodiment, the antibody or fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and / or FR4) consisting of a sequence in any one of SEQ ID NOs: 62-85.

[0122] The antibodies described herein may be defined by their complete light and / or heavy chain variable sequences. Thus, according to a further aspect of the present invention, there is provided an isolated anti-Vδ1 antibody, or a fragment thereof, comprising an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 62 to 85. According to a further aspect of the present invention, there is provided an isolated anti-Vδ1 antibody, or a fragment thereof, consisting of an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 62 to 85.

[0123] In one embodiment, the antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62 to 73. In one embodiment, the antibody or fragment thereof comprises a VH region consisting of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62 to 73. In a further embodiment, the VH region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62, 63, 64, 65, or 66, such as 62, 63, 64, or 65, particularly 62, 63, or 64. In a further embodiment, the VH region consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62, 63, 64, 65, or 66, such as 62, 63, 64, or 65, particularly 62, 63, or 64. In a further embodiment, the VH region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 68, 69, 70, 71, 72, or 73, such as 68, 69, 70, or 71. In a further embodiment, the VH region consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 68, 69, 70, 71, 72, or 73, such as 68, 69, 70, or 71.

[0124] In one embodiment, the antibody or fragment thereof comprises a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74 to 85. In one embodiment, the antibody or fragment thereof comprises a VL region consisting of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74 to 85. In a further embodiment, the VL region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74, 75, 76, 77, or 78, such as 74, 75, 76, or 77, particularly 74, 75, or 76. In a further embodiment, the VL region consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74, 75, 76, 77, or 78, such as 74, 75, 76, or 77, particularly 74, 75, or 76. In a further embodiment, the VL region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 80, 81, 82, 83, 84, or 85, e.g., 80, 81, 82, or 83. In a further embodiment, the VL region consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 80, 81, 82, 83, 84, or 85, e.g., 80, 81, 82, or 83.

[0125] In a further embodiment, the antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62 to 73, and a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74 to 85. In a further embodiment, the antibody or fragment thereof comprises a VH region consisting of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62 to 73, and a VL region consisting of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74 to 85.

[0126] In one embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 63 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 62 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 64 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 68 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 69 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 70 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 71 (1245_P01_G09).

[0127] In one embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 63 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 62 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 64 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 68 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 69 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 70 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 71 (1245_P01_G09).

[0128] In one embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 75 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 74 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 76 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 80 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 81 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 82 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VL region comprising the amino acid sequence of SEQ ID NO: 83 (1245_P01_G09).

[0129] In one embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 75 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 74 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 76 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 80 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 81 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 82 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VL region consisting of the amino acid sequence of SEQ ID NO: 83 (1245_P01_G09).

[0130] In one embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 63 (1252_P01_C08) and a VL region comprising the amino acid sequence of SEQ ID NO: 75 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 62 (1245_P01_E07) and a VL region comprising the amino acid sequence of SEQ ID NO: 74 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 64 (1245_P02_G04) and a VL region comprising the amino acid sequence of SEQ ID NO: 76 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 68 (1139_P01_E04) and a VL region comprising the amino acid sequence of SEQ ID NO: 80 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 69 (1245_P02_F07) and a VL region comprising the amino acid sequence of SEQ ID NO: 81 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 70 (1245_P01_G06) and a VL region comprising the amino acid sequence of SEQ ID NO: 82 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 71 (1245_P01_G06) and a VL region comprising the amino acid sequence of SEQ ID NO: 83 (1245_P01_G09).

[0131] In one embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 63 (1252_P01_C08) and a VL region consisting of the amino acid sequence of SEQ ID NO: 75 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 62 (1245_P01_E07) and a VL region consisting of the amino acid sequence of SEQ ID NO: 74 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 64 (1245_P02_G04) and a VL region consisting of the amino acid sequence of SEQ ID NO: 76 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 68 (1139_P01_E04) and a VL region consisting of the amino acid sequence of SEQ ID NO: 80 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 69 (1245_P02_F07) and a VL region consisting of the amino acid sequence of SEQ ID NO: 81 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 70 (1245_P01_G06) and a VL region consisting of the amino acid sequence of SEQ ID NO: 82 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises a VH region consisting of the amino acid sequence of SEQ ID NO: 71 (1245_P01_G06) and a VL region consisting of the amino acid sequence of SEQ ID NO: 83 (1245_P01_G09).

[0132] For fragments comprising both the VH and VL regions, these may be associated either covalently (e.g., by a disulfide bond or a linker) or non-covalently. The antibody fragments described herein may include scFvs, i.e., fragments comprising the VH and VL regions linked by a linker. In one embodiment, the VH and VL regions are linked by a (e.g., synthetic) polypeptide linker. The polypeptide linker may be (Gly4Ser) nA linker may be included, where n=1 to 8, for example, 2, 3, 4, 5, or 7. The polypeptide linker may be [(Gly4Ser) n (Gly3AlaSer) m ] p It may comprise a linker, where n=1 to 8, for example, 2, 3, 4, 5, or 7, m=1 to 8, for example, 0, 1, 2, or 3, and p=1 to 8, for example, 1, 2, or 3. In a further embodiment, the linker comprises SEQ ID NO: 98. In a further embodiment, the linker consists of SEQ ID NO: 98.

[0133] In one embodiment, the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 86-97. In a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of any one of SEQ ID NOs: 86-97. In yet a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 87 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 86 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 88 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 92 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 93 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 94 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NO: 95 (1245_P01_G09).

[0134] In one embodiment, the antibody or fragment thereof consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 86-97. In a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of any one of SEQ ID NOs: 86-97. In yet a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 87 (1252_P01_C08). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 86 (1245_P01_E07). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 88 (1245_P02_G04). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 92 (1139_P01_E04). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 93 (1245_P02_F07). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 94 (1245_P01_G06). In an alternative embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NO: 95 (1245_P01_G09).

[0135] Those skilled in the art will appreciate that scFv constructs can be designed and engineered to include N- and C-terminal modifications to aid translation, purification, and detection. For example, at the N-terminus of the scFv sequence, additional methionine and / or alanine amino acid residues can be included before the canonical VH sequence (e.g., starting with QVQ or EVQ). At the C-terminus (i.e., C-terminal to the canonical VL domain sequence ending according to the IMGT definition), additional sequences can be included, such as (i) a partial sequence of a constant domain and / or (ii) an additional synthetic sequence, including tags such as His-tags and Flag-tags, to aid purification and detection. In one embodiment, SEQ ID NO: 124 is added to the C-terminus of any one of SEQ ID NOs: 86, 88-90, 92-97. In one embodiment, SEQ ID NO: 125 is added to the C-terminus of any one of SEQ ID NOs: 86, 88-90, 92-97. In one embodiment, SEQ ID NO: 126 is added to the C-terminus of any one of SEQ ID NOs: 87 or 91. In one embodiment, SEQ ID NO: 127 is added to the C-terminus of any one of SEQ ID NOs: 87 or 91. It is well understood that the N- or C-terminal sequences of the scFvs described above are optional and may be removed, modified, or substituted if alternative scFv design, translation, purification, or detection strategies are employed.

[0136] As described herein, antibodies may be in any format. In a preferred embodiment, the antibody is an IgG1 format. Thus, in one embodiment, the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 111-122. In a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of any one of SEQ ID NOs: 111-122. In yet a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NOs: 111-116, e.g., SEQ ID NOs: 111-113 and 116. In yet a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NOs: 117-122, e.g., SEQ ID NOs: 117-120. In yet a further embodiment, the antibody or fragment thereof comprises the amino acid sequence of SEQ ID NOs: 111, 112, 116-120, e.g., SEQ ID NOs: 111, 112, or 116, or SEQ ID NOs: 117-120.

[0137] In one embodiment, the antibody or fragment thereof consists of an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 111-122. In a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of any one of SEQ ID NOs: 111-122. In yet a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NOs: 111-116, e.g., SEQ ID NOs: 111-113 and 116. In yet a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NOs: 117-122, e.g., SEQ ID NOs: 117-120. In yet a further embodiment, the antibody or fragment thereof consists of the amino acid sequence of SEQ ID NOs: 111, 112, 116-120, e.g., SEQ ID NOs: 111, 112 or 116, or SEQ ID NOs: 117-120.

[0138] In one embodiment, the antibody binds to or competes with the same or essentially the same epitope as an antibody or fragment thereof defined herein. Using routine methods known in the art, it is easy to determine whether an antibody binds to the same epitope as a reference anti-V51 antibody or competes with binding thereto. For example, to determine whether a test antibody binds to the same epitope as a reference anti-V51 antibody of the present invention, the reference antibody is allowed to bind to a V51 protein or peptide under saturating conditions. The ability of the test antibody to bind to the V51 chain is then evaluated. If the test antibody can bind to V51 after saturation binding with the reference anti-V51 antibody, it can be concluded that the test antibody binds to a different epitope than the reference anti-V51 antibody. On the other hand, if the test antibody cannot bind to the V51 chain after saturation binding with the reference anti-V51 antibody, the test antibody is likely to bind to the same epitope as the reference anti-V51 antibody of the present invention.

[0139] The present invention also encompasses anti-V51 antibodies that compete for binding to V51 with antibodies or fragments thereof defined herein, or with antibodies having the CDR sequences of any of the exemplary antibodies described herein. For example, competition assays can be performed using the antibodies of the present invention to determine which proteins, antibodies, and other antagonists compete with the antibodies of the present invention for binding to V51 and / or share epitopes. These assays are readily known to those skilled in the art and assess competition between antagonists or ligands for a limited number of binding sites on a protein (e.g., V51). The antibody (or fragment) is immobilized or insolubilized before or after competition, and the sample bound to the V51 chain is separated from the unbound sample, for example, by decantation (if the antibody was pre-insolubilized) or centrifugation (if the antibody was precipitated after the competition reaction). Competitive binding can also be determined by whether its function is altered by the binding or lack of binding of the antibody to the protein, for example, whether the antibody molecule inhibits or enhances, for example, the enzymatic activity of a label. ELISA and other functional assays may be used as known in the art and described herein.

[0140] Two antibodies bind to the same or overlapping epitopes if each competitively inhibits (blocks) the other's binding to a target antigen. That is, a 1-fold, 5-fold, 10-fold, 20-fold, or 100-fold excess of one antibody inhibits the other by at least 50%, but preferably 75%, 90%, or even 99%, as measured in a competitive binding assay. Alternatively, two antibodies have the same epitope if essentially all amino acid mutations in the target antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other.

[0141] Further routine experiments (e.g., peptide mutations and binding analysis) can then be performed to confirm whether the observed loss of binding of the test antibody is indeed due to binding to the same epitope as the reference antibody, or whether steric blocking (or another phenomenon) is responsible for the observed loss of binding. These types of experiments can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art.

[0142] In some embodiments, the antibody or fragment thereof contains an altered effector function through a modification to the sugar linked to Asn 297 (Kabat numbering scheme). In a further modification described above, Asn 297 indicates no or reduced fucosylation (i.e., a defucosylated or nonfucosylated antibody). Fucosylation involves the addition of the sugar fucose to a molecule, for example, the attachment of fucose to N-glycans, O-glycans, and glycolipids. Thus, in a defucosylated antibody, fucose is not attached to the carbohydrate chains of the constant region. An antibody can be modified to prevent or inhibit antibody fucosylation. Typically, glycosylation modification involves expressing the antibody or fragment thereof in a host cell with alternative glycosylation processing capabilities, either through targeted engineering or through targeted or serendipitous host or clonal selection (see, e.g., Example 13). These and other effector modifications are further described in recent reviews, for example, by Xinhua Wang et al. (2018) Protein & Cell 9:63-73 and by Pereira et al. (2018) mAbs 10(5):693-711, which are incorporated herein.

[0143] Antibody sequence modification Antibodies and fragments thereof can be modified using known methods. The sequence modifications to the antibody molecules described herein can be readily incorporated by one of ordinary skill in the art. The following examples are non-limiting.

[0144] During antibody discovery and sequence recovery from phage libraries, desired antibody variable domains can be reformatted into full-length IgGs by subcloning. To accelerate this process, variable domains are often transferred using restriction enzymes. These unique restriction sites may introduce additional / alternative amino acids, deviating from the canonical sequence (such canonical sequences may be found, for example, in the international ImMunoGeneTics [IMGT] information system; see http: / / www.imgt.org). These may be introduced as kappa or lambda light chain sequence modifications.

[0145] Kappa light chain modification The variable kappa light chain variable sequence can be cloned using restriction sites (e.g., Nhe1-Not1) during reformatting to full-length IgG. More specifically, an additional Ala-Ser sequence was introduced at the kappa light chain N-terminus to aid in cloning. Preferably, this additional AS sequence is then removed during further development, such as to generate the canonical N-terminal sequence. Thus, in one embodiment, the kappa light chain-containing antibodies described herein do not contain an AS sequence at their N-terminus; i.e., SEQ ID NOs: 74, 76-78, and 80-85 do not contain the initial AS sequence. In a further embodiment, SEQ ID NOs: 74 and 76-78 do not contain the initial AS sequence. It will be understood that this embodiment also applies to other sequences included herein that contain this sequence (e.g., SEQ ID NOs: 86, 88-90, and 92-97).

[0146] Additional amino acid changes may be made to aid in cloning. For example, for the antibodies described herein, a valine to alanine change was introduced at the kappa light chain variable domain / constant domain boundary to aid in cloning. This resulted in a kappa constant domain modification. Specifically, this results in a constant domain beginning with RTAAAPS (from the NotI restriction site). Preferably, this sequence can be modified during further development to generate a canonical kappa light chain constant region beginning with RTVAAPS. Thus, in one embodiment, the kappa light chain containing antibodies described herein comprise a constant domain described using the sequence RTV. Thus, in one embodiment, the sequence RTAAAPS in SEQ ID NOS: 111-114 and 117-122 is replaced with the sequence RTVAAPS. See, e.g., Example 13 and SEQ ID NOS: 129 and 130.

[0147] Lambda light chain modification Similar to the kappa example above, lambda light chain variable domains may also be cloned by introducing restriction sites (e.g., Nhe1-Not1) during reformatting to full-length IgG. More specifically, an additional Ala-Ser sequence may be introduced at the lambda light chain N-terminus to aid cloning. Preferably, this additional AS sequence is then removed during further development, such as to generate the canonical N-terminal sequence. Thus, in one embodiment, the lambda light chain containing antibodies described herein do not contain an AS sequence at their N-terminus; i.e., SEQ ID NOs: 75 and 79 do not contain the initial AS sequence. It will be understood that this embodiment also applies to other sequences included herein that contain this sequence (e.g., SEQ ID NOs: 87, 91, 115, and 116). In one embodiment, SEQ ID NO: 75 does not contain the first six residues; i.e., the ASSYEL sequence is removed.

[0148] As another example, for the antibodies described herein, a lysine to alanine sequence change was introduced at the lambda light chain variable domain / constant domain boundary to aid in cloning. This resulted in a lambda constant domain modification. Specifically, this resulted in a constant domain starting with GQPAAAPS (from the NotI restriction site). Preferably, this sequence can be modified during further development to generate a canonical lambda light chain constant region starting with GQPKAAPS. Thus, in one embodiment, the lambda light chain containing the antibody described herein comprises a constant domain starting with the sequence GQPK. Thus, in one embodiment, the sequence GQPAAAPS of SEQ ID NO: 115 or 116 is replaced with the sequence GQPKAAPS.

[0149] Heavy chain modifications Typically, human variable heavy chain sequences begin with either basic glutamine (Q) or acidic glutamate (E). However, both of these sequences are known to subsequently be converted to pyroglutamate (pE), an acidic amino acid residue. The Q-to-pE conversion alters the charge on the antibody, while the E-to-pE conversion does not. Therefore, to avoid variable charge changes over time, one option is to modify the starting heavy chain sequence from Q to E in the first instance. Thus, in one embodiment, the heavy chain of an antibody described herein contains a Q to E modification at the N-terminus. In particular, the initial residues of SEQ ID NOs: 62, 64, and / or 67-71 may be modified from Q to E. It will be understood that this embodiment also applies to other sequences contained herein that contain this sequence (e.g., SEQ ID NOs: 86, 88, 91-97, and 111, 112, 115, 117-120). See, for example, Example 13 and SEQ ID NOs:129, 130.

[0150] Furthermore, the C-terminus of the IgG1 constant domain ends with PGK. However, the terminal basic lysine (K) is often subsequently cleaved during expression (e.g., in CHO cells). This results in a change in the charge on the antibody through the variable loss of the C-terminal lysine residue. Therefore, one option is to remove the lysine in the first instance to obtain a uniform and consistent heavy chain C-terminal sequence ending with PG. Thus, in one embodiment, the heavy chain of the antibody described herein has the terminal K removed from its C-terminus. In particular, the antibody of the present invention may comprise any one of SEQ ID NOs: 111 to 122 in which the terminal lysine residue has been removed. See, for example, SEQ ID NO: 141.

[0151] Optional allotypic modification During antibody discovery, specific human allotypes can be used. Optionally, antibodies can be switched to different human allotypes during development. As a non-limiting example, for kappa chains, there are three human allotypes called Km1, Km1,2, and Km3, which define three Km alleles (using allotype numbering). Km1 correlates with valine 153 (IMGT V45.1) and leucine 191 (IMGT L101), Km1,2 correlates with alanine 153 (IMGT A45.1) and leucine 191 (IMGT L101), and Km3 correlates with alanine 153 (IMGT A45.1) and valine 191 (IMGT V101). Therefore, optionally, sequences can be altered from one allotype to another by standard cloning approaches. For example, the L191V (IMGT L101V) change converts the Km1,2 allotype to the Km3 allotype. For further reference to such allotypes, see Jefferis and Lefranc (2009) MAbs 1(4):332-8, which is incorporated herein by reference.

[0152] Thus, in one embodiment, the antibodies described herein contain amino acid substitutions derived from different human allotypes of the same gene. In a further embodiment, the antibodies contain a L191V (IMGT L101V) substitution in the kappa chain to convert the c domain from the km1,2 to km3 allotype. See, e.g., Example 13 and SEQ ID NOs: 129, 130.

[0153] Antibodies targeting epitopes Provided herein are antibodies (or fragments thereof) that bind to an epitope on the V51 chain of a γδ TCR. Such binding may optionally have an effect on γδ TCR activity, such as activation or inhibition.

[0154] In one embodiment, the epitope can be an activating epitope of γδ T cells. An "activating" epitope can include, for example, stimulating TCR function such as cell degranulation, TCR downregulation, cytotoxicity, proliferation, mobilization, increased survival or resistance to exhaustion, intracellular signaling, cytokine or growth factor secretion, phenotypic change, or altered gene expression. For example, binding of an activating epitope can stimulate the expansion (i.e., proliferation) of γδ T cell populations, preferably Vδ1 T cell populations. Thus, these antibodies can be used to regulate γδ T cell activation and thereby regulate immune responses. Thus, in one embodiment, binding of an activating epitope downregulates γδ TCRs. In additional or alternative embodiments, binding of an activating epitope activates γδ T cell degranulation. In further additional or alternative embodiments, binding of an activating epitope activates γδ T cell killing.

[0155] Alternatively, the antibody (or fragment thereof) may have a blocking effect by preventing the binding or interaction of another antibody or molecule. In one embodiment, the invention provides an isolated antibody or fragment thereof that blocks V51 and prevents TCR binding (e.g., by steric hindrance). By blocking V51, the antibody may prevent TCR activation and / or signaling. The epitope may be an inhibitory epitope of γδ T cells. An "inhibitory" epitope may include, for example, blocking TCR function, thereby inhibiting TCR activation.

[0156] In one embodiment, the present invention provides an isolated antibody or fragment thereof that binds to V51 but does not activate γδ T cells (i.e., a non-activating, non-inhibitory, or neutral-binding antibody or fragment thereof), e.g., does not interfere with TCR binding via its "private" CDR3-encoded paratrope. By binding to V51 but not inhibiting or activating γδ T cell activation, in some embodiments, the neutral-binding antibody or fragment thereof can be used to colocalize another molecule. For example, a neutral-binding V51 antibody can be conjugated to a therapeutic moiety such as a cytotoxin or a chemotherapeutic agent. Such a conjugate may be referred to as an immunoconjugate. The antibody can be linked to a cytotoxin, radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, toxin, peptide, or therapeutic agent at any position along the molecule, as long as it is capable of binding to its target. Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins. In one embodiment, the agent can be a second, different antibody directed against V51. In certain embodiments, the antibody may be conjugated to an agent specific for tumor cells or virus-infected cells. The type of therapeutic moiety that may be conjugated to the anti-V51 antibody takes into account the condition to be treated and the desired therapeutic effect to be achieved. In one embodiment, the agent may be a second antibody, or a fragment thereof, that binds to a molecule other than V51.

[0157] The epitope preferably consists of at least an extracellular, soluble, hydrophilic, external or cytoplasmic part of one of the V51 chains of a γδ TCR.

[0158] In particular, the epitope does not include epitopes found in the hypervariable region of the V51 chain of a γδ TCR, particularly in the CDR3 of the V51 chain. In a preferred embodiment, the epitope is within the non-variable region of the V51 chain of a γδ TCR. It will be appreciated that such binding allows for unique recognition of the V51 chain without being limited to the highly variable sequence of the TCR (particularly CDR3). Various γδ TCR complexes that recognize MHC-like peptides or antigens can be recognized in this manner solely by the presence of the V51 chain. It will be appreciated, therefore, that any V51 chain comprising a γδ TCR can be recognized using an antibody or fragment thereof defined herein, regardless of the specificity of the γδ TCR. In one embodiment, the epitope comprises one or more amino acid residues within amino acid regions 1-24 and / or 35-90 of SEQ ID NO: 1, e.g., portions of the V51 chain that are not part of the CDR1 and / or CDR3 sequence. In one embodiment, the epitope does not include amino acid residues within amino acid region 91-105 (CDR3) of SEQ ID NO:1.

[0159] In a manner similar to the well-characterized αβ T cells, γδ T cells utilize a distinct set of somatically rearranged variable (V), diversity (D), joining (J), and constant (C) genes, but γδ T cells contain fewer V, D, and J segments than αβ T cells. In one embodiment, the epitope bound by the antibody (or fragment thereof) does not include an epitope found in the J region of the V51 chain (e.g., one of the four J regions encoded in the human Delta1 chain germline: SEQ ID NO: 152 (J1*0), or 153 (J2*0), or 154 (J3*0), or 155 (J4*0)), or in the C region of the V51 chain (e.g., SEQ ID NO: 156 (C1*0), which contains the C-terminal membrane-proximal / transmembrane region). In one embodiment, the epitope bound by the antibody (or fragment thereof) does not include an epitope found in the N-terminal leader sequence of the V51 chain (e.g., SEQ ID NO: 150). Thus, the antibody or fragment may bind only within the V region of the V51 chain (e.g., SEQ ID NO: 151). Thus, in one embodiment, the epitope consists of an epitope in the V region of a γδ TCR (e.g., amino acid residues 1-90 of SEQ ID NO: 1).

[0160] Reference to the epitope is made in relation to the Vδ1 sequence, which is derived from the sequence described in Luoma et al. (2013) Immunity 39:1032-1042, and shown in SEQ ID NO: 1, RCSB Protein Data Bank entries: 4MNH and 3OMZ. AQKVTQAQSSVSMPVRKAVTLNCLYETSWWSYYIFWYKQLPSKEMIFLIRQGSDEQNAKSGRYSVNFKKAAKSVALTISALQLEDSAKYFCALGESLTRADKLIFGKGTRVTVEPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESS (SEQ ID NO: 1).

[0161] SEQ ID NO: 1 represents a soluble TCR comprising a V region (also referred to as a variable domain), a D region, a J region, and a TCR constant region. The V region comprises amino acid residues 1-90, the D region comprises amino acid residues 91-104, the J region comprises amino acid residues 105-115, and the constant region comprises amino acid residues 116-209. Within the V region, CDR1 is defined as amino acid residues 25-34 of SEQ ID NO: 1, CDR2 is defined as amino acid residues 50-54 of SEQ ID NO: 1, and CDR3 is defined as amino acid residues 93-104 of SEQ ID NO: 1 (Xu et al., PNAS USA 108(6):2414-2419 (2011)).

[0162] Thus, according to an aspect of the present invention, there is provided an isolated antibody or fragment thereof that binds to an epitope of the variable delta 1 (Vδ1) chain of the γδ T cell receptor (TCR) comprising one or more amino acid residues within the following amino acid region: (i) sequences 3 to 20 of SEQ ID NO: 1, and / or (ii) 37 to 77 of SEQ ID NO: 1.

[0163] In a further embodiment, the antibody or fragment thereof further recognizes a polymorphic V region comprising an epitope of amino acid residues 1-90 of SEQ ID NO: 128. Thus, amino acids 1-90 of SEQ ID NO: 1 and the polymorphic germline variant sequence (amino acids 1-90, SEQ ID NO: 128) can be considered interchangeable when defining the epitopes described herein. Studies presented herein demonstrate that the antibodies of the present invention are capable of recognizing both variants of this germline sequence. By way of example, when it is described that an antibody or fragment thereof defined herein recognizes an epitope comprising one or more amino acid residues within amino acid regions 1-24 and / or 35-90 of SEQ ID NO: 1, this also refers to the same regions of SEQ ID NO: 128, specifically amino acid regions 1-24 and / or 35-90 of SEQ ID NO: 128.

[0164] In one embodiment, the antibody or fragment thereof recognizes one or more amino acid residues within amino acid region 1-90 of SEQ ID NO: 1 and amino acids equivalently positioned in region 1-90 of SEQ ID NO: 128. More specifically, in one embodiment, the antibody or fragment thereof defined herein recognizes a human germline epitope, which germline encodes either alanine (A) or valine (V) at position 71 of SEQ ID NO: 1.

[0165] In one embodiment, an epitope includes one or more, eg, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid residues within the described region.

[0166] In a further embodiment, the epitope comprises one or more (e.g., five or more, e.g., ten or more) amino acid residues within amino acid region 3-20 of SEQ ID NO: 1. In an alternative embodiment, the epitope comprises one or more (e.g., five or more, e.g., ten or more) amino acid residues within amino acid region 37-77 (e.g., amino acid region 50-54) of SEQ ID NO: 1. In yet a further embodiment, the epitope comprises one or more (e.g., five or more, e.g., ten or more) amino acid residues within amino acid region 3-20 (e.g., 5-20 or 3-17) and one or more (e.g., five or more, e.g., ten or more) amino acid residues within amino acid region 37-77 (e.g., 62-77 or 62-69) of SEQ ID NO: 1.

[0167] It will be further understood that the above-described antibodies (or fragments thereof) need not bind to all amino acids within a defined range. Such epitopes may be referred to as linear epitopes. For example, an antibody that binds to an epitope comprising amino acid residues within amino acid region 5-20 of SEQ ID NO: 1 may bind only to one or more of the amino acid residues in the range, including, for example, only the amino acid residues at each end of the range (i.e., amino acids 5 and 20), optionally including amino acids within the range (i.e., amino acids 5, 9, 16, and 20).

[0168] In one embodiment, the epitope comprises at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO: 1. In a further embodiment, the epitope comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids selected from amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO: 1.

[0169] In one embodiment, the epitope comprises one or more amino acid residues within the following amino acid region of SEQ ID NO:1 (or SEQ ID NO:128 as described above): (i) 3-17, (ii) 5–20; (iii) 37-53, (iv) 50-64, (v)59-72, (vi) 59-77, (vii) 62-69, and / or (viii) 62-77.

[0170] In a further embodiment, the epitope comprises one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO: 1. In a further embodiment, the epitope consists of one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO: 1.

[0171] In a further embodiment, the epitope comprises, or preferably consists of, amino acid residues 3, 5, 9, 10, 12, 16, 17, 62, 64, 68, and 69 of SEQ ID NO: 1. In a further embodiment, the epitope comprises, or preferably consists of amino acid residues 5, 9, 16, 20, 62, 64, 72, and 77 of SEQ ID NO: 1. In still further embodiments, the epitope comprises, or preferably consists of, amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73, and 77 of SEQ ID NO: 1. In a further embodiment, the epitope comprises, or preferably consists of amino acid residues 50, 53, 59, 62, and 64 of SEQ ID NO: 1. In a further embodiment, the epitope comprises, or preferably consists of amino acid residues 59, 60, 68, and 72 of SEQ ID NO: 1.

[0172] In one embodiment, the epitope comprises one or more amino acid residues within amino acid regions 5-20 and / or 62-77 of SEQ ID NO: 1. In a further embodiment, the epitope consists of one or more amino acid residues within amino acid regions 5-20 and 62-77 of SEQ ID NO: 1. In alternative further embodiments, the epitope comprises one or more amino acid residues within amino acid regions 5-20 or 62-77 of SEQ ID NO: 1. An antibody or fragment thereof having such an epitope may have some or all of the sequence of 1245_P01_E07, or such an antibody or fragment thereof may be derived from 1245_P01_E07. For example, an antibody or fragment thereof having one or more CDR sequences of 1245_P01_E07, or one or both of the VH and VL sequences of 1245_P01_E07, may bind to such an epitope.

[0173] In one embodiment, the epitope comprises one or more amino acid residues within the amino acid region 50-64 of SEQ ID NO: 1. In a further embodiment, the epitope consists of one or more amino acid residues within the amino acid region 50-64 of SEQ ID NO: 1. An antibody or fragment thereof having such an epitope may have some or all of the sequence of 1252_P01_C08, or such an antibody or fragment thereof may be derived from 1252_P01_C08. For example, an antibody or fragment thereof having one or more CDR sequences of 1252_P01_C08, or one or both of the VH and VL sequences of 1252_P01_C08, may bind to such an epitope.

[0174] In one embodiment, the epitope comprises one or more amino acid residues within the amino acid region 37-53 and / or 59-77 of SEQ ID NO: 1. In a further embodiment, the epitope consists of one or more amino acid residues within the amino acid region 37-53 and 59-77 of SEQ ID NO: 1. In an alternative further embodiment, the epitope comprises one or more amino acid residues within the amino acid region 37-53 or 59-77 of SEQ ID NO: 1. An antibody or fragment thereof having such an epitope may have some or all of the sequence of 1245_P02_G04, or such an antibody or fragment thereof may be derived from 1245_P02_G04. For example, an antibody or fragment thereof having one or more CDR sequences of 1245_P02_G04, or one or both of the VH and VL sequences of 1245_P02_G04, may bind to such an epitope.

[0175] In one embodiment, the epitope comprises one or more amino acid residues within the amino acid region 59-72 of SEQ ID NO: 1. In a further embodiment, the epitope consists of one or more amino acid residues within the amino acid region 59-72 of SEQ ID NO: 1. An antibody or fragment thereof having such an epitope may have some or all of the sequence of 1251_P02_C05, or such an antibody or fragment thereof may be derived from 1251_P02_C05. For example, an antibody or fragment thereof having one or more CDR sequences of 1251_P02_C05, or one or both of the VH and VL sequences of 1251_P02_C05, may bind to such an epitope.

[0176] In one embodiment, the epitope does not include amino acid residues in the amino acid region 11-21 of SEQ ID NO: 1. In one embodiment, the epitope does not include amino acid residues in the amino acid region 21-28 of SEQ ID NO: 1. In one embodiment, the epitope does not include amino acid residues in the amino acid region 59 and 60 of SEQ ID NO: 1. In one embodiment, the epitope does not include amino acid residues in the amino acid region 67-82 of SEQ ID NO: 1.

[0177] In one embodiment, the epitope is not the same epitope bound by commercially available anti-V51 antibodies such as TS-1 or TS8.2. As described in WO 2017 / 197347, binding of TS-1 and TS8.2 to soluble TCRs was detected when the 51 chain included the V51 J1 and V51 J2 sequences, but not binding to the V51 J3 chain, indicating that binding of TS-1 and TS8.2 involves critical residues in the deltaJ1 and deltaJ2 regions.

[0178] References herein to "within" include the ends of the defined range. For example, "within amino acid region 5-20" refers to all amino acid residues from residue 5 through residue 20, inclusive.

[0179] Various techniques for determining which epitopes bind to antibodies are known in the art. Exemplary techniques include, for example, routine cross-blocking assays, alanine scanning mutation analysis, peptide blot analysis, peptide truncation analysis, crystallographic studies, and NMR analysis. Additionally, methods such as epitope excision, epitope extraction, and chemical modification of antigens can be used. Another method that can be used to identify amino acids in a polypeptide with which an antibody interacts is hydrogen / deuterium exchange detected by mass spectrometry (as described in Example 9). Generally speaking, the hydrogen / deuterium exchange method involves deuterium-labeling the protein of interest and then binding an antibody to the deuterium-labeled protein. The protein / antibody complex is then transferred to water, and exchangeable protons in amino acids protected by the antibody complex undergo back-exchange from deuterium to hydrogen at a slower rate than exchangeable protons in amino acids that are not part of the interface. As a result, amino acids that form part of the protein / antibody interface can retain deuterium and therefore exhibit a relatively higher mass than amino acids that are not included in the interface. After dissociation of the antibody, the target protein is subjected to protease cleavage and mass spectrometry, revealing deuterium-labeled residues that correspond to the specific amino acids with which the antibody interacts.

[0180] antibody binding The antibody or fragment thereof of the present invention has a solubility of 1.5×10 -7 In a preferred embodiment, the KD is less than 1.5 x 10 -7 In a further embodiment, the KD is less than 1.3 x 10 -7 M (i.e., 130 nM) or less, e.g., 1.0 x 10 -7 In yet a further embodiment, the KD is 5.0 x 10 -8 M (i.e., less than 50 nM), e.g., 4.0 x 10 -8 M (i.e., less than 40 nM), 3.0 × 10 -8M (i.e., 30 nM) or less than 2.0 × 10 -8 M (i.e., 20 nM). For example, according to some embodiments, the concentration is less than 1.5×10 as measured by surface plasmon resonance. -7 Human anti-V51 antibodies are provided that bind to the V51 chain of a γδ TCR with a binding affinity (KD) of less than M (i.e., 150 nM).

[0181] In one embodiment of the invention, 4.0×10 -8 M (i.e., less than 40 nM), 3.0 × 10 -8 M (i.e., 30 nM) or less than 2.0 × 10 -8 Antibodies or fragments thereof that bind to the Vδ1 chain of a γδ TCR with a binding affinity (KD) of less than M (i.e., 20 nM) are provided.

[0182] In one embodiment, the binding affinity of an antibody or fragment thereof is established by coating the antibody or fragment thereof directly or indirectly (e.g., by capturing with anti-human IgG Fc) onto a sensor surface (e.g., an amine high-capacity chip or equivalent), and the target bound by the antibody or fragment (i.e., the V51 chain of the γδ TCR) is flowed over the chip and binding detected. Suitably, a MASS-2 instrument (which may also be referred to as a Sierra SPR-32) is used at 25°C and 30 μL / min in PBS + 0.02% Tween 20 running buffer.

[0183] Other assays that can be used to define antibody function are described herein. For example, an antibody or fragment thereof described herein can be evaluated by measuring γδ TCR engagement, e.g., downregulation of γδ TCR upon antibody binding. Surface expression of γδ TCR (optionally displayed on the surface of cells) after application of the antibody or fragment thereof can be measured, for example, by flow cytometry. An antibody or fragment thereof described herein can also be evaluated by measuring γδ T cell degranulation. For example, expression of CD107a, a marker for cell degranulation, can be measured, for example, by flow cytometry, after application of the antibody or fragment thereof (optionally displayed on the surface of cells) to γδ T cells. An antibody or fragment thereof described herein can also be evaluated by measuring γδ T cell killing activity (to test whether the antibody has an effect on the killing activity of γδ T cells). For example, target cells can be incubated with γδ T cells in the presence of the antibody or fragment thereof (optionally displayed on the surface of cells). After incubation, the culture can be stained with a cell viability dye to distinguish between live and dead target cells. The proportion of dead cells can then be determined, for example, by flow cytometry.

[0184] Drugs for modulating gamma delta T cells The anti-vδ1 antibodies or fragments thereof described herein can be used to modulate delta variable 1 chain (Vδ1) T cells in patients in situ (i.e., in vivo).

[0185] Regulation of V51 T cells may include: - expansion of V51 T cells, e.g., by increasing the number of V51 T cells or promoting the survival of V51 T cells; - stimulation of V51 T cells, for example by increasing the potency of V51 T cells, i.e., by increasing target cell killing; - prevention of Vδ1 T cell exhaustion, e.g., by increasing Vδ1 T cell persistence, - degranulation of Vδ1 T cells, - immunosuppression of V51 T cells, for example by downregulating V51 TCR cell surface expression, i.e. by causing a decrease in V51 TCR internalization or V51 TCR protein expression, or by blocking V51 TCR binding, - Reducing V51 T cell numbers, for example by inhibiting V51 T cell proliferation or by inducing V51 T cell death (i.e. killing V51 T cells).

[0186] Drugs that regulate immune cell markers on Vδ1± cells When administered to a patient, the antibody or fragment thereof may modulate immune cell markers of V51+ cells.

[0187] The suitability of the antibodies or fragments thereof described herein for therapeutic use can also be assessed by measuring γδ T modulation, for example, by measuring changes in the levels of CD25, CD69, or CD107a present on V51 T cells or cells in a model system. Such markers are often used as markers of lymphocyte regulation (e.g., proliferation or degranulation) and can be measured, for example, by flow cytometry, after administration of the antibodies or fragments thereof described herein. Surprisingly, during such evaluations (see, e.g., Examples 7, 17, and 18), it was observed that the antibodies described herein confer measurably higher levels of CD25, CD69, or CD107a to target V51 T cells. Optionally, the phenotypic changes of V51 cells or populations tested in a model system can be compared with the phenotypic changes observed when an alternative comparator antibody (e.g., OKT-3, TS8.2, etc.) is applied to the equivalent γδ T cells described above.

[0188] Thus, in one aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of the γδ TCR for therapeutic use, the method comprising administering the antibody or fragment thereof to a cell population comprising V51+ cells and determining the effect on the levels of CD25 and / or CD69 and / or CD107a on the surface of the V51+ cells. The effect on the levels of CD25, CD69 and / or CD107a may be determined / measured over a period of time. It will be appreciated that the effect can be measured in comparison to the levels of CD25 and / or CD69 and / or CD107a on the surface of V51+ cells when said antibody is not applied to said cells for the same period of time. In a further aspect of the invention, there is provided a method for selecting, characterizing or comparing an antibody or fragment thereof described herein that binds to the V51 chain of the γδ TCR, by applying said antibody to a cell population comprising V51+ cells and then measuring the level (or expression) of CD25 or CD69 or CD107a on the surface of said V51+ cells.

[0189] Drugs that modulate the growth characteristics or number of Vδ1± cells When administered to a patient, the antibody or fragment thereof may modulate the growth properties of V51+ cells, for example, the antibody or fragment thereof may expand V51+ cells.

[0190] An alternative approach for measuring γδ T proliferation may involve measuring the change in the relative number of V51+ cells over time when an antibody or fragment thereof described herein is applied to a model system containing the cells described above. Surprisingly, during such an assessment, it was observed that the antibodies described herein are able to measurably increase the number of V51+ T cells described above (see, e.g., Examples 10, 17, and 18). Optionally, this change in number can then be compared to the change in number observed when an alternative comparator antibody (e.g., anti-OKT3) is applied to the model system described above.

[0191] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, the method comprising administering the antibody or fragment thereof to a cell population comprising V51+ cells and determining the effect on the number of V51+ cells in the population. The effect on cell number may be determined / measured over a period of time. It will be appreciated that the effect may be measured in comparison to the effect on cell number observed when said antibody is not applied to the cell population over the same period of time. In a further aspect of the invention, there is provided a method for selecting, characterizing or comparing antibodies or fragments thereof described herein that bind to the V51 chain of a γδ TCR by applying said antibody to a cell population comprising V51+ cells and then measuring the number of said cells over time.

[0192] Drugs that modulate the proliferative potential and number of Vδ1± cells An ideal therapeutic antibody or fragment thereof described herein that binds to the V51 chain of the γδ TCR would be capable of enhancing V51 cell proliferation in vivo. Such an antibody could then be used as a medicine designed to specifically increase the number of V51+ cells in a subject or patient. For example:

[0193] cancer: A relative increase in the number of V51+ cells has been reported as a positive prognostic indicator associated with improved outcome in many cancers (see, e.g., Gentles et al. (2015) Nature Immunology 21:938-945; Wu et al. (2019) Sci. Trans. Med. 11(513):eaax9364; Catellani et al. (2007) Blood 109(5):2078-2085). In one embodiment, provided herein are medicaments capable of increasing the relative or absolute number of V51+ cells in a system within a cancer patient.

[0194] Pathogenic / parasitic / viral infections: Vδ1 cell enrichment is observed during host defense against numerous acquired pathogenic / parasitic / viral infections. For a recent general review, see Zhao et al. (2018) Immunol. Res. 2018:5081634. Furthermore, increased Vδ1 cell numbers are also thought to be protective against various DNA and RNA viral infections. For example, increased numbers are also considered protective during allogeneic transplant-associated CMV infection (see van Dorp et al. (2011) Biology of Blood and Marrow Transplantation 17(2):S217). Furthermore, Vδ cell numbers increase in patients with coronavirus infection (Poccia et al. (2006) J. Infect. Dis. 193(9):1244-1249).

[0195] In another embodiment, provided herein is a medicament capable of increasing the relative or absolute number of V51+ cells in a subject or patient with a pathogen infection.

[0196] Stem cell transplant: Increased numbers of V51+ cells are also associated with reduced disease relapse during hematopoietic stem cell transplantation, reduced viral infections, increased overall survival and disease-free survival, and generally favorable clinical outcomes (see, e.g., Aruda et al. (2019) Blood 3(21):3436-3448, and Godder et al. (2007) Bone Marrow Transplantation 39:751-757). Accordingly, in another embodiment, presented herein are medicaments that can increase the relative or absolute number of V51+ cells in a subject as part of a therapeutic regimen adjunct to stem cell transplantation.

[0197] Consequently, pharmaceutical agents that can preferentially or specifically increase the number of V51+ cells in a system are highly desirable.

[0198] Drugs that maintain, induce, or increase Vδ1± cell cytokine secretion Cytokines are a large group of proteins, peptides, or glycoproteins secreted by specific cells of the immune system. Cytokines are a category of signaling molecules that mediate and regulate immunity, inflammation, and hematopoiesis. Some cytokines are involved in alleviating the signs and symptoms of disease through either direct or indirect regulation of tumor and cellular microenvironments, autoimmune tissue and related microenvironments, or virally infected tissue or cellular environments. Exemplary pro-inflammatory cytokines include tumor necrosis factor-alpha (TNFα) and interferon-gamma (IFNγ).

[0199] However, many of these cytokines exhibit undesirable toxicities when administered systemically. For example, TNFα can induce hemorrhagic necrosis of transplanted tumors and has been reported to exert synergistic antitumor effects when combined with other chemotherapeutic agents. However, various clinical trials using systemic recombinant human TNFα (rhTNFα) have highlighted significant dose-limiting side effects, including hypotension, rigors, phlebitis, thrombocytopenia, leukopenia, and hepatotoxicity, fever, fatigue, nausea / vomiting, malaise and weakness, headache, chest pain, back pain, diarrhea, and shortness of breath.

[0200] The use of recombinant IFNγ also faces similar systemic toxicity challenges. For example, in cancer settings, IFNγ can exert favorable pleiotropic effects, including upregulation of MHC class I and II to stimulate antitumor immunity, increased T cell infiltration, antiangiogenic effects, induction of chemokine / cytokine secretion, and direct antiproliferative effects on cancer cells. However, adverse side effects are also observed. These adverse side effects include fever, headache, chills, fatigue, diarrhea, nausea, vomiting, loss of appetite, transient increases in hepatic transaminases, and transient decreases in granulocyte and white blood cell counts.

[0201] For a recent review of both the potential and limitations of systemic recombinant TNFα and IFNγ, see Shen et al. (2018) Cell Prolif. 51(4):e12441.

[0202] Therefore, there is a need for more systemically controlled, more localized, and more tissue- or cell-specific production of such cytokines. For example, more controlled expression or induction of pro-inflammatory cytokines has been proposed as one approach that could transform "cold" tumors into "hot" tumors. Highly immunogenic tumors are sometimes referred to as "T cell inflammatory" because they also exhibit increased numbers or densities of CD45+ T cells. For a recent review, see Bonaventura et al. (2019) Front. Immunol. 10:168.

[0203] For this reason, an ideal therapeutic antibody or fragment thereof described herein that binds to the V51 chain of the γδ TCR would be one that is able to maintain or enhance or induce cytokine secretion in V51 cells in vivo. Such an antibody could then be used as a pharmaceutical designed to specifically increase or induce cytokines in a subject or patient in a more localized, less systemic manner, and in a manner that better correlates with the distribution of V51+ cells in said subject or patient.

[0204] Notably, when antibodies described herein that bind to the V51 chain of the γδ TCR are applied to V51+ cells, significantly higher levels of secreted cytokines are observed. More specifically, by way of non-limiting example, significantly higher levels of TNFα and IFNγ are observed. See Example 15.

[0205] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, the method comprising administering the antibody or fragment thereof to a cell population comprising V51 cells and determining the amount of at least one cytokine produced by the cell population. The amount of cytokine produced may be determined / measured over a period of time and, optionally, compared to the amount observed when the antibody is not applied to the cell population over the same period of time. In one embodiment, the observed level of cytokine produced when the antibody is administered to the cell population is about 10% greater, about 20% greater, about 30% greater, about 50% greater, about 100% greater, about 150% greater, about 200% greater, about 250% greater, about 300% greater, about 350% greater, about 400% greater, about 450% greater, about 500% greater, or about 1000% greater than the level of cytokine produced when the antibody is not applied. In a further aspect of the invention, the cytokine is a pro-inflammatory cytokine. In a further aspect of the invention, the cytokine is a TNF-α cytokine. In a further aspect of the invention, the cytokine is an IFN-γ cytokine.

[0206] In a further aspect of the invention there is provided a method for selecting or characterising or comparing an antibody or fragment thereof as described herein which binds to the V51 chain of a γδ TCR, by applying said antibody to a cell population comprising V51+ cells and then measuring the level of at least one cytokine produced. In a further aspect of the invention the cytokine measured is a TNF-α cytokine and / or an IFN-γ cytokine.

[0207] In a further aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of the γδ TCR by applying said antibody or fragment thereof to a cell population comprising V51+ cells and measuring the effect of the antibody on modulating a less or less immunogenic tumor to a more or more immunogenic tumor by determining the amount of pro-inflammatory cytokines produced in the tumor or tumor microenvironment and / or the number or density of CD45+ T cells present in the tumor or tumor microenvironment.

[0208] Drugs that maintain, induce, or increase Vδ1± cell granzyme B activity Granzyme B is a serine protease commonly found in the granules of natural killer cells (NK cells) and cytotoxic T cells. It is secreted by these cells, along with the pore-forming protein perforin, to mediate apoptosis in target cells, including diseased cells.

[0209] When V51+ cells are incubated in co-culture with target disease cells (e.g. cancer cells) in a model system, the level and activity of granzyme B levels can be measured in the target disease cells prior to lysis. Notably, when an antibody or fragment thereof described herein that binds to the V51 chain of the γδ TCR is then applied to such a co-culture of V51+ cells and cancer cells in such a model system, higher granzyme B levels and activity are observed in the disease cancer cells prior to cell death (see Example 16).

[0210] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, the method comprising administering the antibody or fragment thereof to a co-culture comprising V51 cells and disease cells (e.g., cancer cells) and measuring the effect on the amount of granzyme B produced by the disease cells in the co-culture. The amount of cytokine produced may be determined / measured over a period of time and, optionally, compared to the amount observed when the antibody is not applied to the co-culture over the same period of time. In one embodiment, the level of granzyme B measured when the antibody is applied to the co-culture is about 10% greater, about 20% greater, about 30% greater, about 40% greater, about 50% greater, about 70% greater, about 80% greater, about 90% greater, about 100% greater, or about 200% greater than the level of granzyme B observed when the antibody is not applied.

[0211] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or fragment thereof described herein that binds to the V51 chain of a γδ TCR, by applying said antibody to a co-culture comprising V51+ cells and diseased cells and then measuring the amount or activity of granzyme B in the diseased cells.

[0212] Drugs that expand polyclonal Vδ1± cell populations An ideal antibody drug might be one designed to ensure that expanding V51+ cells are not too clonally concentrated at the hypervariable CDR3 sequence level. Thus, an ideal antibody drug could be designed to avoid inducing V51+ cell proliferation by binding to specific or "private" 51+ CDR3 sequence paratropes. Rather, the antibody could bind via a conserved germline sequence present on all V51+ T cell receptors and in a gamma chain-independent manner, rather than binding to a sequence presented on only a subset of V51+ cells.

[0213] Thus, an ideal antibody drug would be able to stimulate the expansion of V51+ cells to generate multiple V51+ cells containing a mixture of CDR3 sequences, resulting in an in vivo expanded heterogeneous polyclonal population of V51+ cells displaying different CDR3 sequences on the delta-variable 1 chain. Notably, during analysis of expanded V51+ cell populations generated by adding the antibodies or fragments thereof described herein to a starting population of immune cells containing V51+ cells, extensive polyclonality is observed using RNAseq-based methodologies designed to sequence through the CDR3 hypervariable region of extracted RNA (see Example 10).

[0214] Thus, in one aspect, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, the method comprising administering the antibody or fragment thereof to a cell population comprising V51 cells and determining the polyclonality of the expanded V51 cells. Desirably, the antibody pharmaceutical generates an expanded polyclonal population that contains multiple V51 CDR3 sequences. Polyclonality can be determined using methods known in the art, such as nucleic acid sequencing approaches that can analyze the V51 chain hypervariable CDR3 content of V51 cells as described above.

[0215] Drugs that expand polyclonal Vδ1± cells over time An ideal antibody drug might be able to enhance, promote, or stimulate the proliferation of primary V51+ cells in vivo without depleting them. For example, by comparison, anti-CD3 drugs such as OKT3 (e.g., muronomab) can expand CD3-positive T cells while also eliminating or inducing anergy. Longer-term proliferation studies were performed to assess the ability of the antibodies described herein that bind to the V51 chain of the γδ TCR to trigger continued cell division of viable V51+ cells. Remarkably, these studies revealed that the antibodies described herein that bind to the V51 chain of the γδ TCR can trigger cell division / proliferation of viable and still functionally cytotoxic V51+ cells for over 40 days (see Example 10).

[0216] In one embodiment, a method is provided for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, comprising applying the antibody or fragment thereof to a cell population and monitoring the length of time that V51+ cell division occurs. Ideally, the antibody is able to stimulate V51+ cell division over a period of 5 to 60 days, for example, at least 7 to 45 days, 7 to 21 days, or 7 to 18 days.

[0217] In a further embodiment there is provided an antibody or fragment thereof as described herein which binds to the V51 chain of a γδ TCR and which, when administered to a patient, is capable of stimulating V51+ cell division and increasing their number by at least 2-fold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 100-fold, at least 200-fold, at least 300-fold, at least 400-fold, at least 500-fold, at least 600-fold, at least 1,000-fold.

[0218] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or fragment thereof described herein that binds to the V51 chain of a γδ TCR, by applying said antibody to V51+ cells or a mixed cell population containing V51+ cells and then measuring the number of V51+ cells over time.

[0219] Drugs that modulate non-Vδ1± immune cells by targeting Vδ1± immune cells The antibodies or fragments thereof described herein can also be evaluated by measuring V51+ cell-mediated regulation of other immune cells. For example, changes observed in the non-γδ T cell "fraction" can be measured after applying the antibodies or fragments thereof described herein to a model system containing a mixed population of immune cells, such as those containing human tissue αβ cells and γδ T cells. Furthermore, the effect on non-γδ cell types in the above-mentioned model can be measured by flow cytometry. For example, by measuring the relative change in CD8+ αβ T cell numbers when the antibodies or fragments thereof described herein are added to a mixed culture containing γδ T cells and non-γδ T cells. Optionally, the observed change in the number or phenotype of non-γδ T cell CD8+ lymphocyte populations can then be compared to the change in numbers when a surrogate comparator antibody (e.g., OKT-3) is applied to the above-mentioned mixed population.

[0220] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, comprising administering the antibody or fragment thereof to a mixed population of immune cells or tissues comprising V51+ cells and V51-negative immune cells and measuring the effect on V51-negative immune cells. The effect may be determined / measured over a period of time and, optionally, compared to the effect observed in V51-negative cells when the antibody is not applied over the same period of time. The effect may be measured as a change in the number of V51-negative immune cells. For example, the antibody may increase the number of V51-negative immune cells by more than about 10%, more than about 20%, more than about 30%, more than about 40%, more than about 50%, more than about 70%, more than about 80%, more than about 90%, more than about 100%, or more than about 500% compared to the level observed when the antibody is not applied.

[0221] In a further aspect of the invention, the regulated V51-negative cells are CD45+ cells. In a further aspect of the invention, the regulated cells are αβ T cells. In a further aspect of the invention, the regulated αβ+ cells are CD8+ lymphocytes. In a further aspect of the invention, the regulated αβ T cells, or a population thereof, show evidence of enhanced cell division. In a further aspect of the invention, there is provided a method for selecting, characterizing or comparing an antibody or fragment thereof described herein that binds to the V51 chain of the γδ TCR, by administering said antibody to a population of mixed immune cells comprising V51+ cells and V51-negative immune cells, and then measuring the effect on the V51-negative cell population by V51+ cells regulated by said antibody or fragment thereof.

[0222] Optionally, during the "V51+ cell-mediated immune system modulation" effected by the antibodies or fragments thereof described herein, a concomitant increase in the number of V51+ cells is also observed. Also, without being bound by this theory, it is possible that the increase in the number of V51+ cells described above is responsible for promoting the simultaneous expansion of co-existing V51-negative immune cells, such as αβ T cells. An alternative hypothesis may be that antibody-induced cytokine secretion from V51+ T cells stimulates the expansion of V51-negative immune cells.

[0223] In a further aspect of the invention, the increase observed in the αβ+CD8+ lymphocyte population is compared to a comparator antibody, such as the OKT3 antibody or an alternative anti-V51 antibody. In a further aspect of the invention, there is provided a method of selecting or characterizing or comparing antibodies or fragments thereof described herein that bind to the V51 chain of the γδ TCR, by applying the above-mentioned antibodies to a population of mixed immune cells comprising V51+ T cells and αβ T cells and then measuring the number of CD8+αβ+ T cell lymphocytes over time.

[0224] Drugs that regulate tumor-infiltrating lymphocytes (TIL) The antibodies or fragments thereof described herein can also be evaluated by measuring the effect on tumor-infiltrating corpuscles (TILs) in a model system. Surprisingly (see Example 18), during such evaluation, the antibodies described herein measurably modulated TIL corpuscles in human tumors. For example, changes in either the number or phenotype of γδ+ lymphocyte TIL corpuscles or non-γδ lymphocyte TIL corpuscles are measured after applying the antibodies or fragments thereof described herein to human tumors, such as human renal cell carcinoma. Optionally, the observed changes in either the number or phenotype of γδ+ lymphocyte TIL corpuscles or non-γδ lymphocyte TIL corpuscles can then be compared with the changes observed when an alternative comparator antibody (e.g., OKT-3) is applied to the above-mentioned model system.

[0225] Thus, in another aspect of the present invention, a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR is provided, comprising administering the antibody or fragment thereof to TILs located in or derived from a human tumor and determining the effect on the number of TILs. The effect may be determined / measured over a period of time and, optionally, compared to the number of TILs observed when the antibody is not administered over the same period of time. The effect may be an increase in the number of TILs. For example, the antibody may increase the number of TILs by about 10%, about 20%, about 30%, about 40%, about 50%, about 70%, about 80%, about 90%, or about 100% compared to the number of TILs observed when the antibody is not administered. In a further aspect, the number of TILs observed is γδ+ lymphocyte TIL cells and / or non-γδ lymphocyte TIL cells.

[0226] In a further aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or fragment thereof as described herein that binds to the V51 chain of a γδ TCR cell antibody by applying said antibody to a TIL or TILs located in or derived from a human tumor and then measuring the change in the number of TIL or TIL cells over a period of time.

[0227] Drugs that modulate human Vδ1± cytotoxicity The antibodies or fragments thereof described herein can also be evaluated by measuring the effect they have on V51+-mediated cytotoxicity. Surprisingly, during such evaluation of the antibodies described herein (see, e.g., Example 19), measurably enhanced V51+-mediated cytotoxicity was observed. For example, after applying the antibody or fragment thereof to a model system comprising a mixed culture containing V51+ cells and the above-mentioned cancer cells, a decrease in the number of cancer cells or an increase in the number of killed cancer cells is observed. Optionally, the decrease in the number of cancer cells or the increase in the number of killed cancer cells can then be compared with the results when an alternative comparator antibody (e.g., OKT-3) is applied to the above-mentioned model system.

[0228] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, comprising applying the antibody or fragment thereof to a mixed population of cells comprising V51 cells and cancer cells, and measuring the cytotoxicity of the V51 cells against the cancer cells. Cytotoxicity may be measured by the increase in the number of dead cancer cells over a period of time, optionally compared to the number of dead cancer cells observed when the antibody is not applied to the mixed population of cells over the same period of time. For example, the increase in dead cells observed when the antibody is applied may be greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 70%, greater than about 80%, greater than about 90%, greater than about 100%, greater than about 200%, or greater than about 500% relative to the number of dead cells observed when the antibody is not applied.

[0229] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or fragment thereof as described herein that binds to the V51 chain of γδ TCR, by adding said antibody to the above-mentioned collection of mixed immune cells comprising human V51+ cells and cancer cells, and then measuring the increase in dead cancer cells over time.

[0230] Drugs that modulate the Vδ1± cell target-to-effector cell ratio (T:E ratio) The antibodies or fragments thereof described herein may also be evaluated by measuring how they enhance V51+-mediated cancer cell cytotoxicity by determining the target cell-to-effector cell ratio resulting in 50% killing of target cells (EC50) in a model system, thereby evaluating the antibody as a potential therapeutic. For example, a mixed culture comprising target cancer cells and human V51+ effector cells. Surprisingly, during such evaluation (see, e.g., Example 19), the antibodies described herein desirably alter the EC50 T:E ratio in the model system. Such alteration can be measured as the number of V51+ cells required to observe 50% killing of cancer cells over a set period of time. This can also be reported as a change in cytotoxicity against the cancer cells, or as a fold or percentage improvement. Optionally, the T:E ratio provided by the antibodies of the present invention can then be compared to the T:E ratio when an alternative comparator antibody (e.g., OKT-3) is applied to the model system.

[0231] Thus, in another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, comprising applying the antibody or fragment thereof to a mixed population of cells comprising human V51 cells and cancer cells, and measuring the number of V51 cells required to kill 50% of the cancer cells. This may optionally be measured in comparison with the number of V51 cells required to kill 50% of the cancer cells over the same period of time without application of the antibody. For example, the reduction in the number of V51 cells required to kill 50% of the cancer cells when the antibody is applied can be greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 70%, greater than about 80%, greater than about 90%, greater than about 100%, greater than about 200%, or greater than about 500% compared to the number of V51 cells required to kill 50% of the cancer cells when the antibody is not applied.

[0232] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or fragment thereof described herein that binds to the V51 chain, by adding said antibody to an above-mentioned collection of cells comprising V51+ cells and cancer cells, and then measuring the number of V51+ cells required to kill 50% of the cancer cells.

[0233] Drugs that enhance EC50 cytotoxicity of Vδ1± cells An alternative way to measure the observed enhanced cytotoxicity of human V51+ cells or populations thereof is to measure the number of cells required to kill 50% of the cancer cells over a period of time in condition A (e.g., starting control) and compare this to the number of cells required to kill 50% of the cancer cells over a period of time in condition B (e.g., upon application of an antibody of the invention as described herein).

[0234] It is recognized that there are a variety of ways to measure such parameters, but to aid in understanding, the following non-limiting hypothetical example is outlined.

[0235] Hypothetically, the enhancement of effector cell cytotoxicity can be measured as follows: - In condition A (control treatment), it is observed that 1000 V51+ cells are required to kill 50% of the cancer cells over a given period of time (e.g., 5 hours). - In condition B (e.g., application of an antibody of the invention as described herein), it is observed that 500 V51+ cells are required to kill 50% of the cancer cells over the same period of time. Thus, in this example, application of the antibody has enhanced the cytotoxicity of the V51+ cell population by 200%. (1000 / 500) x 100 = 200%

[0236] For example (see Example 19), such enhancement rates are surprisingly observed for the antibodies of the invention described herein.

[0237] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or fragment thereof as described herein that binds to the V51 chain of a γδ TCR, by adding said antibody to said collection of mixed immune cells comprising V51+ cells and cancer cells and determining the relative change or percent change in cytotoxicity compared to an equivalent or control experiment in which said antibody is not applied to said mixture of cells.

[0238] A drug that enhances Vδ1± cell specificity to diseased cells while sparing healthy cells Another approach to evaluating the antibodies or fragments thereof described herein is to measure how the antibodies modulate disease cell-specific cytotoxicity. Surprisingly, during such studies, it was discovered that such antibodies specifically enhance Vδ1 cell-specific killing of disease cells, such as cancer cells (see, e.g., Example 19), while sparing healthy or non-disease cells. An ideal antibody drug administered to patients to alleviate cancer symptoms would provide enhanced cytotoxicity specifically against disease cells, while sparing healthy cells. Drugs that enhance effector cell cytotoxicity specifically against disease cells, such as cancer cells, can be said to exhibit an enhanced therapeutic index (TI) compared with drugs that do not selectively enhance effector cytotoxicity specifically against the aforementioned disease cells. The therapeutic index, also referred to as the therapeutic ratio, is a quantitative measure of a drug's relative safety. It is the comparison between the amount of a therapeutic agent that causes a therapeutic effect and the amount that causes toxicity, for example, by causing unwanted death in related or relevant healthy cell populations. The antibodies or fragments thereof described herein can be evaluated by measuring their ability to alter, enhance, or fold-improve the ability of V51+ cells to selectively kill diseased cells over and above healthy cells in a model system. For example, the model system described above can include V51+ effector cells, cancer cells, and control cells (e.g., healthy cells). Optionally, the fold improvement in selective diseased cell killing provided by the antibodies of the invention can then be compared to the fold improvement observed when an alternative comparator antibody (e.g., OKT-3) is applied to the model system described above.

[0239] The disease cell specificity and enhanced disease cell specificity of V51+ cells can be measured in cultures containing V51+ cells, diseased cells, and healthy cells. For example, V51+ specificity for diseased cells can be measured by observing the number of cancer cells killed by V51+ cells and then comparing the number of healthy cells killed by V51+ cells. Such comparisons can be controlled by including equal numbers of diseased and healthy cells in a model system that also contains V51+ cells (e.g., "triculture"). Alternative comparison methodologies can also be considered, for example, when analytical or instrumental limitations limit the ability to distinguish and track three or more cell types in parallel in a single assay (including V51+ cells, diseased cells, and non-disease cells). In the example above, comparing V51+ cytotoxicity against diseased cells in one experiment and then V51+ cytotoxicity against healthy cells in separate, equivalent experiments provides an alternative approach to such studies.

[0240] In another aspect of the invention, there is provided a method for evaluating an antibody or fragment thereof that binds to the V51 chain of a γδ TCR, the method comprising administering the antibody or fragment thereof to a cell population comprising V51 cells and target cells, and measuring the cytotoxic specificity of the cell against the target cells. In one embodiment, the cytotoxic specificity of the cell against a first target cell type can be compared to the cytotoxicity observed against a second target cell type, and the method may be repeated using a different target cell type. In a further aspect of the invention, the first target cell type is a diseased cell, and the second target cell type is a control cell, such as a healthy cell or a cell with a disease different from the first target cell type.

[0241] In a further aspect of the invention, there is provided a method for selecting or characterizing or comparing antibodies or fragments thereof described herein that bind to the V51 chain of a γδ TCR, wherein the effect caused by said antibodies on the cytotoxicity of V51+ cells against (i) a first cell type and (ii) a second cell type is measured and compared. In a further aspect of the invention, the antibodies are selected such that their specific cytotoxicity against the first cell type is enhanced relative to their specific cytotoxicity against the second cell type. In a further aspect of the invention, the first cell type is a diseased cell and the second cell type is a healthy cell.

[0242] As described herein, the antibody or fragment thereof used in the assay can be displayed on a surface, for example, the surface of a cell, such as a cell containing an Fc receptor. For example, the antibody or fragment thereof can be displayed on the surface of a THP-1 cell, such as a TIB-202™ cell (available from the American Type Culture Collection (ATCC)). Alternatively, the antibody or fragment thereof can be used directly in the assay.

[0243] In such functional assays, output can be measured by calculating the half maximal effective concentration, also referred to as "EC50" or "effective concentration at 50%." The term "IC50" refers to inhibitory concentration. Both EC50 and IC50 can be measured using methods known in the art, such as flow cytometry. In some cases, EC50 and IC50 can be considered to be the same value or equivalent. For example, the effective concentration (EC) of effector cells required to inhibit (e.g., kill) 50% of a particular cell type can also be considered the 50% inhibitory concentration (IC). For the avoidance of doubt, EC50 values ​​in this application, when referring to antibodies, are provided using IgG1-formatted antibodies. Such values ​​can be readily converted based on the molecular weight of the antibody format for equivalent values ​​as follows: (μg / ml) / (MW in kDa) = μM

[0244] The EC50 for down-regulation of γδ TCR upon antibody (or fragment) binding may be less than 0.50 μg / ml, for example, less than 0.40 μg / ml, less than 0.30 μg / ml, less than 0.20 μg / ml, less than 0.15 μg / ml, less than 0.10 μg / ml, less than 0.06 μg / ml, or less than 0.05 μg / ml. In a preferred embodiment, the EC50 for down-regulation of γδ TCR upon antibody (or fragment) binding is less than 0.10 μg / ml. In particular, the EC50 for down-regulation of γδ TCR upon antibody (or fragment) binding may be less than 0.06 μg / ml, for example, less than 0.05 μg / ml, less than 0.04 μg / ml, or less than 0.03 μg / ml. In particular, the above-mentioned EC50 values ​​are when the antibody is measured in IgG1 format. For example, EC50 γδ TCR down-modulation values ​​can be measured using flow cytometry (eg, as described in the assay in Example 6).

[0245] The EC50 for γδ T cell degranulation upon antibody (or fragment) binding may be less than 0.050 μg / ml, for example, less than 0.040 μg / ml, less than 0.030 μg / ml, less than 0.020 μg / ml, less than 0.015 μg / ml, less than 0.010 μg / ml, or less than 0.008 μg / ml. In particular, the EC50 for γδ T cell degranulation upon antibody (or fragment) binding may be less than 0.005 μg / ml, for example, less than 0.002 μg / ml. In a preferred embodiment, the EC50 for γδ T cell degranulation upon antibody (or fragment) binding is less than 0.007 μg / ml. In particular, the above-mentioned EC50 values ​​are when the antibody is measured in IgG1 format. For example, γδ T cell degranulation EC50 values ​​can be measured by detecting CD107a expression (i.e., a marker of cell degranulation) using flow cytometry (e.g., as described in the assay in Example 7). In one embodiment, CD107a expression is measured using an anti-CD107a antibody, such as anti-human CD107a BV421 (clone H4A3) (BD Biosciences).

[0246] The EC50 for γδ T cell killing upon antibody (or fragment) binding may be less than 0.50 μg / ml, for example, less than 0.40 μg / ml, less than 0.30 μg / ml, less than 0.20 μg / ml, less than 0.15 μg / ml, less than 0.10 μg / ml, or less than 0.07 μg / ml. In a preferred embodiment, the EC50 for γδ T cell killing upon antibody (or fragment) binding is less than 0.10 μg / ml. In particular, the EC50 for γδ T cell killing upon antibody (or fragment) binding may be less than 0.060 μg / ml, for example, less than 0.055 μg / ml, particularly less than 0.020 μg / ml. In particular, the above-mentioned EC50 values ​​are those when the antibody is measured in IgG1 format. For example, an EC50 γδ T cell killing value can be measured by detecting the percentage of dead cells (e.g., using a cell viability dye) using flow cytometry after incubation of the antibody, γδ T cells, and target cells (e.g., as described in the assay in Example 8). In one embodiment, target cell death is measured using a cell viability dye, where the viability dye is Viability Dye eFluor™ 520 (ThermoFisher).

[0247] In the assays described in these embodiments, the antibody or fragment thereof can be displayed on the surface of cells such as THP-1 cells, e.g., TIB-202™ (ATCC). The THP-1 cells are optionally labeled with a dye such as CellTracker™ Orange CMTMR (ThermoFisher).

[0248] Immunoconjugates The antibodies or fragments thereof of the present invention may be conjugated to a therapeutic moiety such as a cytotoxin or a chemotherapeutic agent. Such conjugates may also be referred to as immunoconjugates. As used herein, the term "immunoconjugate" refers to an antibody that is chemically or biologically linked to another moiety, such as a cytotoxin, radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, toxin, peptide or protein, or therapeutic agent. The antibody may be linked to the cytotoxin, radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, toxin, peptide, or therapeutic agent at any position along the molecule, as long as it is capable of binding to its target. Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins. In one embodiment, the agent may be a second, different antibody against V51. In certain embodiments, the antibody may be conjugated to an agent specific for tumor cells or virus-infected cells. The type of therapeutic moiety that may be conjugated to an anti-V51 antibody takes into account the condition to be treated and the desired therapeutic effect to be achieved. In one embodiment, the agent may be a second antibody, or a fragment thereof, that binds to a molecule other than V51.

[0249] multispecific antibodies Antibodies of the present invention may be monospecific, or they may bind to additional targets and thus be bispecific or multispecific. Multispecific antibodies may be specific for different epitopes of a single target polypeptide, or may be specific for more than one target polypeptide. Thus, in one embodiment, the antibody or fragment thereof comprises a first binding specificity for V51 and a second binding specificity for a second target epitope.

[0250] In various embodiments, the second target epitope is an epitope of a cancer antigen or a cancer-associated antigen, such as AFP, AKAP-4, ALK, alpha-fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCAA, Bcr-abl, beta-catenin, beta-HCG, beta-human chorionic gonadotropin, BORIS, BTAA, CA 125, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, CAM. 17.1, CAM43, carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33 / IL3Ra, CD68 / P1, CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG4), c-Met, CO-029, CSPG4, cyclin B1, cyclophilin C-related protein, CYP1B1, E2A-PRL, EGFR, EGFRvIII, ELF2M, EpCAM, EphA2, ephrin B2, Epstein-Barr virus antigen EBVA, ERG (TMPRSS2ETS fusion gene), ETV6-AML, FAP, FGF-5, Fos-related antigen 1, fucosyl GM1, G250, Ga733 / EpCAM, GAGE-1, GAGE-2, GD2, GD3, glioma-associated antigen, GloboH, glycolipid F77, GM3, GP 100, GP 100 (Pmel 17), H4-RET, HER-2 / neu, HER-2 / Neu / ErbB-2, high molecular weight melanoma-associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin growth factor (IGF)-I, intestinal carboxylesterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, legumain, LMP2, M344, MA-50, Mac-2 binding protein, MAD-CT-1, MAGE, MAGE A1, MAGE A3, MAGE-1, MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1, MART-1 / MelanA, M-CSF, melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin, MG7-Ag, ML-IAP, MN-CAIX, MOV18, MUC1, Mum-1, hsp70-2, MYCN, MYL-RAR, NA17, NB / 70K, neuron-glial antigen 2 (NG2), neutrophil elastase, nm-23H1, NuMa, NY-BR-1, NY-CO-1, NY-ESO, NY-ESO-1, NY-ESO-1, OY-TES1, p15, p16, p180erbB3, p185erbB2, p53, p53 mutant, Page4, PAX3, PAX5, PDGFR-based and TA, PLAC1, polysialic acid, prostate carcinoma tumor antigen-1 (PCTA-1), prostate-specific antigen, prostatic acid phosphatase (PAP), proteinase 3 (PR1), PSA, PSCA, PSMA, RAGE-1, Ras, Ras-mutant, RCAS1, RGS5, RhoC, ROR1, RU1, RU2 (AS), SART3, SDCCAG16, sLe(a), sperm protein 17, SSX2, STn, survivin, TA-90, TAAL6, a TAG-72, telomerase, thyroglobulin, Tie 2, TIGIT, TLP, Tn, TPS, TRP-1, TRP-2, TRP-2, TSP-180, tyrosinase, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72.

[0251] In various embodiments, the second target epitope is an epitope of a cluster of differentiation CD antigen, hi various embodiments, the CD antigens are CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CD13, CD14, CD15, CD16, CD16a, CD16b, CD17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32A , CD32B, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49a, CD 49b, CD49c, CD49d, CD49e, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60a, CD60b, CD60c, CD61, CD62E, CD62L, CD62P, C D63, CD64a, CD65, CD65s, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD75s, CD77, CD79A, CD79B , CD80, CD81, CD82, CD83, CD84, CD85A, CD85B, CD85C, CD85D, CD85F, CD85G, CD85H, CD85I, CD85J, CD85K, CD85M, CD86, CD87, CD88, CD89, CD90, CD9 1, CD92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107, CD107a, CD107b, CD108, CD109, CD11 0, CD111, CD112, CD113, CD114, CD115, CD116, CD117, CD118, CD119, CD120, CD120a, CD120b, CD121a, CD121b, CD122, CD123, CD124, CD125, CD126,CD127、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140A、CD140B、CD141、CD142、CD143、CD144、CDw145、CD146、CD147、CD148、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD157、CD158、CD158A、CD158B1、CD158B2、CD158C、CD158D、CD158E1、CD158E2、CD158F1、CD158F2、CD158G、CD158H、CD158I、CD158J、CD158K、CD159a、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CDw198、CDw199、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218a、CD218b、CD219、CD220、CD221、CD222、CD223、CD224、CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240CE、CD240D、CD241、CD242、CD243、CD244、CD245

[17] 、CD246、CD247、CD248、CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256, CD257, CD258, CD259, CD260, CD261, CD262, CD263, CD264, CD265, CD266, CD267, CD268, CD269, CD 270, CD271, CD272, CD273, CD274, CD275, CD276, CD277, CD278, CD279, CD280, CD281, CD282, CD283, CD284 , CD285, CD286, CD287, CD288, CD289, CD290, CD291, CD292, CDw293, CD294, CD295, CD296, CD297, CD298, C D299, CD300A, CD300C, CD301, CD302, CD303, CD304, CD305, CD306, CD307, CD307a, CD307b, CD307c, CD307 d, CD307e, CD308, CD309, CD310, CD311, CD312, CD313, CD314, CD315, CD316, CD317, CD318, CD319, CD320, CD321, CD322, CD323, CD324, CD325, CD326, CD327, CD328, CD329, CD330, CD331, CD332, CD333, CD334, CD3 35, CD336, CD337, CD338, CD339, CD340, CD344, CD349, CD351, CD352, CD353, CD354, CD355, CD357, CD358, CD360, CD361, CD362, CD363, CD364, CD365, CD366, CD367, CD368, CD369, CD370, and CD371.

[0252] Although the mechanisms by which γδ T cells recognize antigens and distinguish between healthy and diseased cells are not fully understood (Ming Heng and Madalene Heng, Antigen Recognition by γδ T-Cells. Madame Curie Bioscience Database [Internet], Austin (TX): Landes Bioscience; 2000-2013), the fact that γδ T cells can distinguish between healthy and diseased cells and exhibit significant disease cell multicytotoxicity (non-limiting examples of cell types in Table 1) means that they can be harnessed to provide improved medicines with improved therapeutic windows. Furthermore, exploiting this ability of γδ T cells offers the opportunity to treat disease while sparing healthy cells by colocalizing γδ T cells with diseased cells, even when specific cancer, inflammatory, or pathogen antigens are unknown or are also present on healthy cells in a particular patient. JPEG2025160218000002.jpg216167

[0253] As one non-limiting example, recent studies using CD3×HER2 polyspecifics highlight the challenges of current or conventional approaches. Specifically, the use of such conventional approaches can result in unfavorable toxicity profiles. This is because, like many other tumor-associated antigens (TAAs), the HER2 antigen is expressed not only in cancers such as breast cancer but also in healthy tissues such as cardiac cells. Therefore, the use of CD3×HER2 drugs that engage and colocalize all T cells with HER2-positive cells may result in a less favorable therapeutic window or therapeutic index. This is because such drugs engage all T cells in the circulation, the majority of which are αβ T cells (CD4+ positive, CD8+ positive, etc.). Furthermore, when αβ T cells colocalize with HER2-positive cells, such conventional αβ T cells exhibit limited ability to spare HER2+ healthy cells and limited ability to kill only HER2+ disease cells. As a result, according to this example, in cynomolgus monkey studies administered such CD3xHER2 bispecific drugs, early euthanasia (even on the day of administration) was necessary in some circumstances. Furthermore, during this example study (see Staflin et al. (2020) JCI Insight 5(7):e133757), it was concluded that retargeting T cells to kill HER2-expressing cells can induce adverse effects on HER2-expressing tissues. It was noted that all diseased or damaged tissues express HER2, except for the liver.

[0254] In a further non-limiting example, the second binding specificity may be directed against a tumor-associated moiety that is also involved in the control or regulation of immune cell function. For example, the second specificity may be designed to target so-called "checkpoint inhibitors," such as PD-L1 (CD274) or CD155. Again, neither PDL-1 nor CD155 is 100% disease-specific. Both proteins can also be expressed on healthy cells. However, multispecific antibodies designed to specifically colocalize Vδ1+ cells with either PD-L1- or CD155-positive cells can cause selective killing of PD-L1- or CD155-positive disease or cancer cells. Further targeting of disease-associated checkpoint inhibitors present on diseased cells, such as cancer cells, may not only co-localize V51+ cells to such tumors, but may also confer additional positive effects, for example, by modulating or attenuating PD-1 / PD-L1 or TIGIT / CD155 signaling that may otherwise negatively regulate T cell-mediated immune responses to disease.

[0255] Therefore, instead of using such conventional approaches, multispecific antibodies are provided herein that can bind to V51+ cells via at least one first binding specificity and to targets present on diseased tissues and cells via at least one second binding specificity. Use of such multispecific antibodies in this manner can result in colocalization of V51+ cells with diseased cells expressing the second target. Furthermore, because a given disease-associated target is often not 100% disease-specific, this approach of specifically targeting and colocalizing V51+ effector cells may be preferable to conventional approaches. This is because V51+ effector cells can recognize stress patterns in diseased or infected cells, thereby selectively killing diseased cells while sparing healthy cells expressing the same target.

[0256] In a further non-limiting example, a patient may have liver cancer for which the liver cancer-specific antigen is unknown. In this case, the second specificity of the multispecific antibody may be directed against an epitope present on many or all liver cells, such as asialoglycoprotein receptor 1. This then allows γδ T cells to co-localize with the liver, allowing the γδ T cells to kill liver cancer cells while sparing healthy liver cells. As a third non-limiting example, in a patient with lung cancer for which the lung cancer antigen is unknown, the second specificity of the multispecific antibody may be directed against an epitope on normal lung cells, such as SP-1. This allows γδ T cells to co-localize with the lung, allowing the γδ T cells to kill lung cancer cells while sparing healthy lung cells. As a fourth non-limiting example, in a patient with B cell lymphoma for which the B cell lymphoma antigen is unknown, the second specificity of the multispecific antibody may be directed against an epitope on normal B cells, such as CD19. This allows the γδ T cells to co-localize with B cells, allowing the γδ T cells to kill lymphoma cells while sparing healthy B cells. Cell-specific, cell-associated, tissue-specific, and tissue-associated antigens are well known in the art, and any such antigen may be targeted by the second specificity of the multispecific antibody of the invention.

[0257] The second binding specificity may target an antigen on the same cell as V51, or on a different cell of the same tissue type, or on a different cell of a different tissue type. In certain embodiments, the target epitopes may be on different cells, including different T cells, B cells, tumor cells, autoimmune tissue cells, or virus-infected cells. Alternatively, the target epitopes may be on the same cell.

[0258] Multispecific antibodies or fragments thereof can be produced in any format, so long as the antibody or fragment thereof has multiple specificities. Examples of multispecific antibody formats include, but are not limited to, CrossMab, DAF (two-in-one), DAF (four-in-one), DutaMab, DT-IgG, knob-in-hole (KIH), knob-in-hole (common light chain), charge pair, Fab arm exchange, SEED body, triomab, LUZ-Y, Fcab, κλ-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L,H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, and KIH. IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG (Four-in-One), Nanobody, Nanoby-HAS, BiTE, Diabody, DART, TandAb, sc diabody, sc diabody-CH3, diabody-CH3, triple body, mini antibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFV-CH-CL-scFv, F(ab')2, F(ab;)2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, sc diabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock, ImmTAC, HSA body, sc diabody-HAS, tandem scFv-toxin, IgG-IgG, ov-X-Body, duobody, mab 2 , and scFv1-PEG-scFv2 (see Spiess et al. (2015) Molecular Immunology 67:95-106).

[0259] The antibodies or fragments thereof described herein may also be evaluated by measuring their ability for enhanced function in multispecific formats, such as bispecific or trispecific formats. Surprisingly, through such studies, it is possible to identify even further functional improvements in the performance of the antibodies or fragments thereof described herein (see Examples 20 and 21).

[0260] A variety of antibody-derived multispecific formats have been described before, and are typically constructed empirically from component binding moieties.Typically, once constructed, the performance of such multispecific or multitarget binding formats described herein can be measured in one or more of the above-mentioned model systems (cell killing, cell proliferation, healthy cell spared / disease cell specific model, etc.).Optionally, they are also compared with the above-mentioned component moieties and other comparator molecules.

[0261] Generally, without being limited to this approach, when antibodies are constructed as multispecific antibodies, binding domain modules for each target (first, second, third, etc.) are optionally constructed from scFvs, Fabs, Fab's, F(ab')2s, Fvs, variable domains (e.g., VH or VL), diabodies, minibodies, or full-length antibodies. For example, each of the above-mentioned binding domains or modules is produced in one or more of the following non-limiting formats, in which binding domains comprising variable domains, and / or full-length antibodies, and / or antibody fragments are operably linked in series to generate the multispecific antibody.

[0262] Notably, multispecific antibodies comprising at least one (first) binding domain targeting the V51 chain of the γδ TCR described herein are further enhanced when said first binding domain is formatted together with a multispecific antibody format comprising at least one second binding domain directed against any tissue ("solid") and hematopoietic ("liquid") disease or cell type associated target.

[0263] Multispecific antibodies - non-limiting examples: To outline the applicability of the approach, a series of non-limiting example multispecific antibodies were constructed that comprise at least one (first) binding domain that targets the V51 chain of the γδ TCR and at least one (second) binding domain that targets a disease-associated target.

[0264] First example; Vδ1-EGFr multispecific antibody: In this example, one binding domain (against a first target) comprised an intact antibody moiety, specifically VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (against a second target) comprised an antibody fragment, specifically an scFv format. A linker was then utilized to fuse the two binding modules. The resulting bispecific format is sometimes referred to as the "Morrison format." In this case, the first binding domain targets the V51 chain of the γδ TCR, and the second binding domain targets EGFr (see Example 20).

[0265] Second example; Vδ1-EGFr multispecific antibody: For example, one binding domain (against a first target) comprises antibody variable domains (specifically comprising a VH and cognate VL domain), while the second binding domain (against a second target) comprises binding domains within the heavy chain constant domains (CH1-CH2-CH3) (see Table 1 of EP2546268 A1 / EP3487885 A1). The resulting bispecific comprises a first binding domain that targets the V51 chain of the γδ TCR and a second binding domain that targets the EGF receptor (see Example 20).

[0266] Third example; Vδ1-CD19 multispecific antibody: In this example, one binding domain (against a first target) comprised an intact antibody moiety, specifically comprising VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (against a second target) comprised an antibody fragment, specifically comprising an scFv format. A linker was then utilized to fuse the two binding modules. In this example, the resulting bispecific comprised a first binding domain targeting the V51 chain of the γδ TCR and a second binding domain targeting CD19 (see Example 21).

[0267] Notably, in all of the above examples comprising at least one (first) binding domain targeting the V51 chain of the γδ TCR, at least one second domain targeting a second epitope was observed to have enhanced functionality relative to the control and component moieties (see Examples 20 and 21 herein).

[0268] Taken together, these non-limiting examples highlight the flexibility of the antibodies or fragments thereof described herein. These non-limiting examples outline a multispecific antibody approach in which an antibody, fragment thereof, targeting the germline Vδ1 chain (amino acids 1-90 of SEQ ID NO: 1), can be further enhanced by combining it with a second binding domain to form a multispecific antibody. As a non-limiting example, multispecific antibodies with enhanced function are provided herein containing binding domains comprising intact antibodies (VH-CH1-CH2-CH3 and VL-CL), and / or variable domains (VH and cognate VL or VH-CH1 and cognate VL-CL), and / or antibody fragments (scFv).

[0269] In one embodiment, a multispecific antibody binding domain targeting the V51 chain of the γδ TCR (first target) may comprise: (i) one or two or more antibody binding domains, each comprising a heavy chain (VH-CH1-CH2-CH3) and a cognate light chain partner (VL-CL); and / or (ii) one or two or more antibody binding domains, each comprising a heavy chain variable domain (VH, or VH-CH1) and a cognate light chain variable domain partner (VL, or VL-VC); and / or (iii) one or two or more antibody binding domains, each comprising a CDR-containing antibody fragment.

[0270] In one embodiment, a multispecific antibody is provided, comprising at least one binding domain derived from a first antibody targeting the V51 chain of the γδ TCR, operably linked to at least one second antibody binding domain targeting a second epitope. Optionally, the binding domains comprise at least one or more VH and cognate VL binding domains, or one or more VH-CH1-CH2-CH3 and cognate VL-CL binding domains, or one or more antibody fragment binding domains. Optionally, the second binding domain targets a second epitope associated with or expressed on the cell surface of a cell. Optionally, the second epitope is located on a cell surface polypeptide associated with diseased or tumor cells, or virally infected or autoimmune tissue cells. Optionally, the second epitope or epitopes are located on a disease- and cell-type-associated CD19 or EGFr antigen. Optionally, said multispecific antibody comprising at least one first antibody-derived binding domain targeting the V51 chain of the γδ TCR is operably linked to a second binding domain that binds to the EGF receptor and comprises one or more of the following heavy chain modifications according to EU nomenclature: L358T and / or T359D and / or K360D and / or N361G and / or Q362P and / or N384T and / or G385Y and / or Q386G and / or D413S and / or K414Y and / or S415W and / or Q418Y and / or Q419K.

[0271] Optionally, the multispecific antibody comprises at least one first antibody-derived binding domain targeting the V51 chain of the γδ TCR, comprising SEQ ID NO: 147 or SEQ ID NO: 148 or SEQ ID NO: 149 or SEQ ID NO: 157 or a functionally equivalent binding variant thereof, operably linked to a second binding domain targeting either EGFr or CD19. Optionally, the resulting multispecific antibody comprises SEQ ID NO: 140 or SEQ ID NO: 141 or SEQ ID NO: 142 or SEQ ID NO: 144 or SEQ ID NO: 145 or SEQ ID NO: 146 or SEQ ID NO: 158 or SEQ ID NO: 159. The above-mentioned entities are included herein as non-limiting novel compositions to aid in understanding.

[0272] In one aspect of the invention, the multispecific antibodies of the invention may be used in a therapeutically effective amount to treat a disease or disorder so as to alleviate at least one sign or symptom of the disease or disorder.

[0273] In one embodiment, there is provided a method for selecting or characterizing or comparing antibodies or fragments thereof described herein that bind to the V51 chain of a γδ TCR in a multispecific antibody format, wherein said multispecific antibody is applied to V51+ cells in order to measure the effect exerted by said multispecific entity V51+ cells (e.g. on said V51+ phenotype and / or cytotoxicity and / or disease cell specificity and / or enhancement thereof).

[0274] Polynucleotides and Expression Vectors In one aspect of the present invention, there is provided a polynucleotide encoding an anti-V51 antibody or multispecific antibody, or fragment thereof, of the present invention. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to SEQ ID NOs: 99-110. In one embodiment, an expression vector comprises a VH region of SEQ ID NOs: 99-110. In another embodiment, an expression vector comprises a VL region of SEQ ID NOs: 99-110. In a further embodiment, the polynucleotide comprises or consists of SEQ ID NOs: 99-110. In a further aspect, there is provided a cDNA comprising the above-described polynucleotide.

[0275] In one aspect of the present invention, a polynucleotide encoding an anti-Vδ1 antibody or fragment of the present invention is provided. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to SEQ ID NOs: 99-101 or 105-108. In one embodiment, an expression vector comprises a VH region of SEQ ID NOs: 99-101 or 105-108. In another embodiment, an expression vector comprises a VL region of SEQ ID NOs: 99-101 or 105-108. In a further embodiment, the polynucleotide comprises or consists of SEQ ID NOs: 99-101 or 105-108. In a further aspect, a cDNA comprising the above-described polynucleotide is provided.

[0276] In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, for example at least 90%, such as at least 95%, for example at least 99% sequence identity to SEQ ID NOs: 99-101. In one embodiment, the expression vector comprises a VH region of SEQ ID NOs: 99-101. In another embodiment, the expression vector comprises a VL region of SEQ ID NOs: 99-101. In a further embodiment, the polynucleotide comprises or consists of SEQ ID NOs: 99-101. In a further aspect, there is provided a cDNA comprising the above-mentioned polynucleotide.

[0277] In one aspect of the invention, there is provided a polynucleotide comprising or consisting of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to any one of a portion of SEQ ID NOs: 99-110 encoding CDR1, CDR2 and / or CDR3 of the encoded immunoglobulin chain variable domain. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to any one of a portion of SEQ ID NOs: 99-101 or 105-108 encoding CDR1, CDR2 and / or CDR3 of the encoded immunoglobulin chain variable domain. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, for example at least 90%, such as at least 95%, for example at least 99% sequence identity to any one of the portions of SEQ ID NOs: 99-101 encoding CDR1, CDR2 and / or CDR3 of the encoded immunoglobulin chain variable domain.

[0278] In one aspect of the invention, there is provided a polynucleotide comprising or consisting of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to any one of a portion of SEQ ID NOs: 99-110 encoding FR1, FR2, FR3 and / or FR4 of the encoded immunoglobulin chain variable domain. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity to any one of a portion of SEQ ID NOs: 99-101 or 105-108 encoding FR1, FR2, FR3 and / or FR4 of the encoded immunoglobulin chain variable domain. In one embodiment, the polynucleotide comprises or consists of a sequence having at least 70%, such as at least 80%, for example at least 90%, such as at least 95%, for example at least 99% sequence identity to any one of the portions of SEQ ID NOs: 99-101 encoding FR1, FR2, FR3 and / or FR4 of the encoded immunoglobulin chain variable domain.

[0279] Polynucleotides and expression vectors of the invention may also be described with reference to the encoded amino acid sequence. Thus, in one embodiment, a polynucleotide comprises or consists of a sequence encoding the amino acid sequence of any one of SEQ ID NOs: 62-85. In one embodiment, an expression vector comprises a sequence encoding the amino acid sequence of any one of SEQ ID NOs: 62-73. In another embodiment, an expression vector comprises a sequence encoding the amino acid sequence of any one of SEQ ID NOs: 74-85.

[0280] To express an antibody or fragment thereof, polynucleotides encoding the partial or full-length light and heavy chains described herein are inserted into an expression vector such that the genes are operably linked to transcriptional and translational control sequences. Accordingly, one aspect of the present invention provides an expression vector comprising a polynucleotide sequence defined herein. In one embodiment, the expression vector comprises a VH region of SEQ ID NO: 99-110, e.g., SEQ ID NO: 99, 100, 101, 105, 106, 107, or 108. In another embodiment, the expression vector comprises a VL region of SEQ ID NO: 99-110, e.g., SEQ ID NO: 99, 100, 101, 105, 106, 107, or 108.

[0281] It is understood that the nucleotide sequences described herein include additional sequences encoding amino acid residues to aid in translation, purification, and detection; however, alternative sequences may be used depending on the expression system used. For example, the first (5'-end) 9 nucleotides of SEQ ID NOs: 99-110, the last (3'-end) 36 nucleotides of SEQ ID NOs: 99-100, 102-103, and 105-110, or the last (3'-end) 39 nucleotides of SEQ ID NOs: 101 and 104 are optional sequences. These optional sequences can be removed, modified, or substituted if alternative design, translation, purification, or detection strategies are employed.

[0282] Mutations can be made to the DNA or cDNA encoding a polypeptide that are silent with respect to the amino acid sequence of the polypeptide but provide preferred codons for translation in a particular host. For example, preferred codons for translation of nucleic acids in E. coli and S. cerevisiae, as well as mammals, particularly humans, are known.

[0283] Mutations in polypeptides can be achieved, for example, by substitutions, additions, or deletions to the nucleic acid encoding the polypeptide. Substitutions, additions, or deletions to the nucleic acid encoding the polypeptide can be introduced by a number of methods, including, for example, error-prone PCR, shuffling, oligonucleotide-directed mutagenesis, assembly PCR, PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive ensemble mutagenesis, exponential ensemble mutagenesis, site-specific mutagenesis, gene reassembly, artificial gene synthesis, gene site saturation mutagenesis (GSSM), synthetic ligation reassembly (SLR), or a combination of these methods. Modifications, additions, or deletions to nucleic acids can also be introduced by methods including recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, uracil-containing template mutagenesis, gapped duplex mutagenesis, point mismatch repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiolytic decay mutagenesis, deletion mutagenesis, restriction-selection mutagenesis, restriction-purification mutagenesis, ensemble mutagenesis, chimeric nucleic acid multimer generation, or a combination thereof.

[0284] In particular, artificial gene synthesis can be used. Genes encoding the polypeptides of the present invention can be synthetically produced, for example, by solid-phase DNA synthesis. The entire gene can be synthesized de novo without the need for precursor template DNA. To obtain the desired oligonucleotide, building blocks are sequentially attached to a growing oligonucleotide chain in the order required by the product sequence. Once chain assembly is complete, the product is released from the solid phase into solution, deprotected, and collected. The product can be isolated by high-performance liquid chromatography (HPLC) to obtain the desired oligonucleotide in high purity.

[0285] Expression vectors include, for example, plasmids, retroviruses, cosmids, yeast artificial chromosomes (YACs), and Epstein-Barr virus (EBV)-derived episomes. A polynucleotide is ligated into a vector such that transcriptional and translational control sequences within the vector perform their intended function of regulating the transcription and translation of the polynucleotide. Expression and / or control sequences may include promoters, enhancers, transcription terminators, a start codon (i.e., ATG) 5' to the coding sequence, splicing signals for introns, and stop codons. Expression vectors and expression control sequences are selected to be compatible with the expression host cell used. SEQ ID NOs: 99-110 include nucleotide sequences encoding single-chain variable fragments of the invention, comprising VH and VL regions connected by a synthetic linker (encoding SEQ ID NO: 98). It will be understood that the polynucleotides or expression vectors of the invention may comprise a VH region, a VL region, or both (optionally including a linker). Thus, polynucleotides encoding the VH and VL regions can be inserted into separate vectors, or alternatively, sequences encoding both regions are inserted into the same expression vector. Polynucleotides are inserted into expression vectors by standard methods (e.g., ligation of complementary restriction sites on the polynucleotide and vector, or blunt-end ligation if no restriction sites are present).

[0286] A convenient vector is one that encodes a functionally complete human CH or CL immunoglobulin sequence with appropriate restriction sites engineered to allow for the easy insertion and expression of any VH or VL sequence, as described herein. Expression vectors can also encode a signal peptide that facilitates secretion of the antibody (or fragment thereof) from the host cell. A polynucleotide can be cloned into the vector such that the signal peptide is linked in-frame to the amino terminus of the antibody. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

[0287] In one aspect of the present invention, a cell (e.g., a host cell) is provided that contains a polynucleotide or expression vector as defined herein.It will be understood that the cell may contain a first vector encoding the light chain of an antibody or its fragment, and a second vector encoding the heavy chain of an antibody or its fragment.Alternatively, both the heavy chain and the light chain are encoded on the same expression vector introduced into the cell.

[0288] In one embodiment, the polynucleotide or expression vector encodes a membrane anchor or transmembrane domain fused to an antibody or fragment thereof, such that the antibody or fragment thereof is displayed on the extracellular surface of the cell.

[0289] Transformation can be by any known method for introducing polynucleotide into host cell.The method for introducing heterologous polynucleotide into mammalian cell is well known in the art, and includes dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of polynucleotide in liposome, biolistic injection and direct microinjection of DNA into nucleus.In addition, nucleic acid molecule can be introduced into mammalian cell by virus vector.

[0290] Mammalian cell lines available as expression hosts are well known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC). These cell lines include, among others, Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney (COS) cells, human hepatocellular carcinoma cells (e.g., HepG2), A549 cells, 3T3 cells, and several other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cow, horse, and hamster cells. A particular preferred cell line is selected by determining which cell line has a high expression level. Other cell lines that can be used are insect cell lines such as Sf9 cells, amphibian cells, bacterial cells, plant cells, and fungal cells. Antigen-binding fragments of antibodies, such as ScFv and Fv fragments, can be isolated and expressed in E. coli using methods known in the art.

[0291] The antibody is produced by culturing the host cells for a period of time sufficient to allow expression of the antibody in the host cells, or more preferably, secretion of the antibody into the medium that the host cells are grown in. The antibody can be recovered from the culture medium using standard protein purification methods.

[0292] The antibodies (or fragments) of the present invention can be obtained and engineered using, for example, the techniques disclosed in Green and Sambrook, Molecular Cloning: A Laboratory Manual (2012) 4th Edition, Cold Spring Harbour Laboratory Press.

[0293] Monoclonal antibodies can be produced using hybridoma technology by fusing specific antibody-producing B cells with myeloma (B-cell cancer) cells selected for the ability to grow in tissue culture and the absence of antibody chain synthesis.

[0294] Monoclonal antibodies directed against the determined antigens can be prepared, for example, by Antibodies can be obtained by: a) immortalizing lymphocytes obtained from the peripheral blood of an animal previously immunized with a determined antigen with immortal cells, preferably myeloma cells, to form hybridomas; b) culturing the formed immortalized cells (hybridomas) and recovering the cells that produce antibodies with the desired specificity.

[0295] Alternatively, the use of hybridoma cells is not necessary. Antibodies capable of binding to the target antigens described herein can be isolated from suitable antibody libraries through conventional practices, for example, using phage display, yeast display, ribosome display, or mammalian display techniques known in the art. Thus, monoclonal antibodies can be isolated, for example, from a) cloning into a vector, in particular a phage, more particularly a filamentous bacteriophage, DNA or cDNA sequence obtained from lymphocytes, in particular peripheral blood lymphocytes, of an animal (preferably previously immunized with the determined antigen); b) transforming a prokaryotic cell with the vector described above under conditions that allow the production of the antibody; c) selecting antibodies by performing antigen affinity selection; and d) recovering antibodies having the desired specificity.

[0296] Optionally, isolated polynucleotides encoding the antibodies or fragments thereof described herein and binding to the V51 chain of gamma delta can also be readily produced in sufficient quantities for use as pharmaceuticals to ameliorate signs or symptoms of disease. When used as pharmaceuticals in this manner, typically, the polynucleotide of interest is first operably linked to an expression vector or expression cassette designed to express the antibody or fragment thereof in a subject or patient. Such expression cassettes and methods for delivering polynucleotides, or sometimes referred to as "nucleic acid-derived" drugs, are well known in the art. For a recent review, see Hollevoet and Declerck (2017) J. Transl. Med. 15(1):131.

[0297] Pharmaceutical Compositions According to a further aspect of the present invention, there is provided a composition comprising an antibody or fragment thereof as defined herein. In such embodiments, the composition may comprise the antibody, optionally in combination with other excipients. Also included are compositions comprising one or more additional active agents (e.g., active agents suitable for treating the diseases referred to herein).

[0298] According to a further aspect of the present invention, a pharmaceutical composition is provided comprising an antibody or fragment thereof defined herein together with a pharmaceutically acceptable diluent or carrier. The antibody of the present invention can be incorporated into a pharmaceutical composition suitable for administration to a subject. Typically, the pharmaceutical composition comprises the antibody of the present invention and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonicity agents, and absorption delaying agents. Examples of pharmaceutically acceptable carriers include one or more of water, saline, salt, phosphate-buffered saline, dextrose, glycerol, ethanol, etc., and combinations thereof. In many cases, it is preferable to include an isotonicity agent, such as a sugar, a polyalcohol such as mannitol, sorbitol, or sodium chloride, in the composition. Pharmaceutically acceptable substances, such as wetting or minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives, or buffers, that enhance the shelf life or effectiveness of the antibody or fragment thereof, are also included.

[0299] The compositions of the present invention can be in various forms.These compositions include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories.Preferred forms depend on intended administration modes and therapeutic applications.Typical preferred compositions are in the form of injectable and infusible solutions.

[0300] The preferred mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular, intrathecal). In a preferred embodiment, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

[0301] Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration.

[0302] It is within the scope of the present invention to use the pharmaceutical compositions of the present invention in therapeutic methods for the treatment of the diseases described herein as an adjunct to, or in conjunction with, other established therapies commonly used to treat such diseases.

[0303] In a further embodiment of the invention, the antibody, composition or pharmaceutical composition is administered sequentially, simultaneously or separately with at least one active agent.

[0304] Treatment method According to a further aspect of the present invention, there is provided an isolated anti-V51 antibody or fragment thereof as defined herein for use as a pharmaceutical. As used herein, reference to an antibody or fragment thereof "for use" as a pharmaceutical or in therapy is limited to administration of the antibody or fragment thereof to a subject. Such use does not include administering the antibody or fragment thereof to a cell culture (i.e., in vitro or ex vivo), where the cell culture or derived cell therapy product is used as a therapeutic agent.

[0305] In one embodiment, the anti-V51 antibody or fragment thereof is for use in treating cancer, an infectious disease, or an inflammatory disease. In one embodiment, the invention is a method of treating a disease or disorder in a subject in need thereof, comprising administering an anti-V51 antibody or fragment thereof to the subject. In various embodiments, the disease or disorder is cancer, an infectious disease, or an inflammatory disease. In one embodiment, the anti-V51 antibody or fragment thereof is for use in treating cancer, an infectious disease, or an inflammatory disease that causes the death of diseased cells while sparing healthy cells. In a further embodiment, the antibody or fragment thereof is for use in treating cancer.

[0306] In one embodiment, the antibody or fragment thereof is for use in the treatment of cancer, an infectious disease, or an inflammatory disease, hi a further embodiment, the antibody or fragment thereof is for use in the treatment of cancer.

[0307] According to a further aspect of the present invention, there is provided a pharmaceutical composition as defined herein for use as a medicament. In one embodiment, the pharmaceutical composition is for use in the treatment of cancer, an infectious disease, or an inflammatory disease. In a further embodiment, the pharmaceutical composition is for use in the treatment of cancer.

[0308] According to a further aspect of the invention, there is provided a method of modulating an immune response in a subject in need thereof, the method comprising administering a therapeutically effective amount of an isolated anti-V51 antibody or fragment thereof as defined herein. In various embodiments, modulating the immune response in the subject comprises binding or targeting γδ T cells, activating γδ T cells, causing or increasing γδ T cell proliferation, causing or increasing γδ T cell expansion, causing or increasing γδ T cell degranulation, causing or increasing γδ T cell killing activity, causing or increasing γδ T cell killing activity while sparing healthy cells, causing or increasing γδ T cytotoxicity, causing or increasing γδ T cytotoxicity while sparing healthy cells, causing or increasing γδ T cell recruitment, increasing γδ T cell survival, or increasing resistance to γδ T cell depletion.

[0309] According to a further aspect of the invention there is provided a method of treating cancer, an infectious disease or an inflammatory disease in a subject in need thereof, comprising administering a therapeutically effective amount of an isolated anti-V51 antibody or fragment thereof as defined herein. Alternatively, a therapeutically effective amount of a pharmaceutical composition is administered.

[0310] According to a further aspect of the invention there is provided the use of an antibody or fragment thereof as defined herein for the manufacture of a medicament, for example in the treatment of cancer, an infectious disease or an inflammatory disease.

[0311] In one embodiment, the antibody or fragment thereof is administered to a subject, wherein the subject has cancer, an infectious disease, or an inflammatory disease.

[0312] According to a further aspect of the present invention there is provided a pharmaceutical composition as defined herein for use as a medicament, in one embodiment the pharmaceutical composition is administered to a subject having cancer, an infectious disease or an inflammatory disease.

[0313] According to a further aspect of the invention, there is provided a method of administering a therapeutically effective amount of an isolated anti-V51 antibody or fragment thereof as defined herein to a subject, wherein the subject has cancer, an infectious disease or an inflammatory disease. Alternatively, a therapeutically effective amount of a pharmaceutical composition is administered.

[0314] According to a further aspect of the invention there is provided the use of an antibody or fragment thereof as defined herein for the manufacture of a medicament, e.g., for administration to a subject, wherein the subject has cancer, an infectious disease or an inflammatory disease.

[0315] In various embodiments, cancers treatable by the methods and compositions of the present disclosure include, but are not limited to, acute lymphoblastic, acute myeloid leukemia, adrenocortical carcinoma, appendix cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, osteosarcoma, and malignant fibrous histiocytoma, brain stem glioma, brain tumor, central nervous system atypical teratoid rhabdoid tumor, central nervous system embryonal tumor, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, craniopharyngioma, ependymoblastoma, ependymoma, medulloblastoma, medulloepithelioma, intermediate pineal parenchymal tumor, epiprimitive neuroectodermal tumor, and pineoblastoma, Gliomas of the visual pathway and hypothalamus, tumors of the brain and spinal cord, breast cancer, bronchial tumors, Burkitt's lymphoma, carcinoid tumors, gastrointestinal carcinoid tumors, atypical teratoid rhabdoid tumors of the central nervous system, embryonal tumors of the central nervous system, central nervous system lymphomas, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, esophageal cancer, Ewing's family of tumors, extragonadal germ cell tumors, extrahepatic bile duct cancer, intraocular melanoma, retinoblastoma, Gallbladder cancer, gastric cancer (gastric (stomach) cancer), gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (gist), germ cell tumor, gestational trophoblastic tumor, glioma, brain stem glioma, cerebral astrocytoma glioma, visual pathway and hypothalamic glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, Langerhans cell histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway glioma, intraocular melanoma, islet cell tumor, kidney (renal cell) cancer, Langerhans cell histiocytosis, laryngeal cancer, acute lymphoblastic leukemia, acute myeloma Myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, lip and oral cavity cancer, liver cancer, non-small cell lung cancer, small cell lung cancer, AIDS-related lymphoma, Burkitt's lymphoma, cutaneous T-cell lymphoma, non-Hodgkin's lymphoma, primary central nervous system lymphoma, Waldenstrom's macroglobulinemia, malignant fibrous histiocytoma and osteosarcoma of bone, medulloblastoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell neck cancer of unknown primary, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis, mycosis fungoides, myelodysplastic syndrome,Myelodysplastic / myeloproliferative disorders, myeloid leukemia, myeloid leukemia, acute myeloid leukemia, multiple myeloma, myeloproliferative disorders, cancer of the nasal cavity and paranasal sinuses, nasopharyngeal carcinoma, neuroblastoma, non-small cell lung cancer, oral cancer, oral cavity cancer, oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, intermediate pineal parenchymal tumor, pineoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter, respiratory tract carcinoma with nut gene on chromosome 15 These include retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Ewing's family of tumors, Kaposi's sarcoma, soft tissue sarcoma, uterine sarcoma, Sézary syndrome, skin cancer (non-melanoma), skin cancer (melanoma), Merkel cell skin carcinoma, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell cervical cancer, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumor, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, gestational trophoblastic tumor, urethral cancer, endometrial cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, and Wilms' tumor. In various embodiments, the methods and compositions of the present disclosure treat treatable cancers while sparing healthy cells.

[0316] In various embodiments, inflammatory diseases that can be treated by the methods and compositions of the present disclosure include, but are not limited to, achalasia, acute disseminated encephalomyelitis (ADEM), acute motor axonal neuropathy, acute respiratory distress syndrome (ARDS), Addison's disease, steatohepatitis dolorosa, adult-onset Still's disease, adult-onset Still's disease, agammaglobulinemia, alopecia areata, amyloidosis, amyotrophic lateral sclerosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, anti-N-methyl D-aspartate (anti-NMDA) receptor encephalitis, antiphospholipid syndrome, antiphospholipid syndrome (APS), APLS), antisynthetase syndrome, antiglomerular basement membrane nephritis, aplastic anemia, atopic allergy, atopic dermatitis, autoimmune angioedema, autoimmune complications, autoimmune autonomic dysfunction, autoimmune encephalomyelitis, autoimmune enteropathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis (AIP), autoimmune peripheral neuropathy, autoimmune polyendocrine syndrome (APS) type 1, autoimmune polyendocrine syndrome Autoimmune polyendocrine syndrome (APS) type 2, autoimmune polyendocrine syndrome (APS) type 3, autoimmune retinopathy, autoimmune thrombocytopenic purpura, autoimmune thyroiditis, autoimmune urticaria, autoimmune uveitis, autoimmune vasculitis, axonal neuronal neuropathy (AMAN), Baro concentric sclerosis, Baro disease, Behçet's disease, benign mucous membrane pemphigoid, Bickerstaff encephalitis, Blau syndrome, bullous pemphigoid, Castleman disease (CD), celiac disease, Chagas disease, chronic fatigue syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic obstructive pulmonary disease, chronic relapsing multiple sclerosis Critically Ill Osteomyelitis (CRMO), Churg-Strauss Syndrome (CSS) or Eosinophilic Granulomatosis (EGPA), Pemphigoid, Cogan's Syndrome, Cold Agglutinin Disease, Complement Component 2 Deficiency, Complex Regional Pain Syndrome, Congenital Heart Block, Connective Tissue, Systemic and Multiorgan, Contact Dermatitis, Coxsackie Myocarditis, Crest Syndrome, Crohn's Disease, Cushing's Syndrome, Cutaneous Leukocytoclastic Vasculitis, Degos' Disease, Dermatitis Herpetiformis, Dermatomyositis, Devic's Disease (Neuromyelitis Optica), Type 1 Diabetes, Gastrointestinal System, Discoid Lupus, Dressler's Syndrome, Drug-Induced Lupus, Eczema, Endometriosis, Enthesitis-Associated Arthritis,Eosinophilic esophagitis (EoE), eosinophilic fasciitis, eosinophilic gastroenteritis, eosinophilic granulomatosis with polyangiitis (EGPA), eosinophilic pneumonia, epidermolysis bullosa acquisita, erythema nodosum, erythroblastosis fetalis, esophageal achalasia, essential mixed cryoglobulinemia, Evans syndrome, exocrine glands, Felty syndrome, fibrodysplasia ossificans progressiva, fibromyalgia, fibrosing alveolitis, gastritis, gastrointestinal pemphigoid, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture syndrome syndrome, granulomatosis with polyangiitis, Graves' ophthalmopathy, Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis, Hashimoto's encephalopathy, hemolytic anemia, Henoch-Schönlein purpura (HSP), herpes gestationis or pemphigoid of gestationis (PG), hidradenitis suppurativa (HS) (acne inversa), hypogammaglobulinemia, idiopathic giant cell myocarditis, idiopathic inflammatory demyelinating disease, idiopathic pulmonary fibrosis, IgA neuropathy, IgA vasculitis (IgAV), IgG4-related disease, IgG4-related sclerosis inflammatory bowel disease, immune thrombocytopenic purpura (ITP), inclusion body myositis (IBM), inflammatory bowel disease, intermediate uveitis, interstitial cystitis (IC), interstitial lung disease, IPEX syndrome, juvenile arthritis, juvenile diabetes mellitus (type 1 diabetes), juvenile myositis (JM), Kawasaki disease, Lambert-Eaton syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, lignified conjunctivitis, linear immunoglobulin A disease (LAD), lupus, lupus nephritis, lupus vasculitis, chronic Lyme disease, Majeed syndrome, Nierre's disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's ulcer, morphea, Mukka-Habermann disease, multifocal motor neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myocarditis, myositis, narcolepsy, neonatal lupus, nervous system, neuromyelitis optica, neuromyotonia, neutropenia, ocular cicatricial pemphigoid, opsoclonus-myoclonus syndrome, optic neuritis, Ord's thyroiditis thyroiditis), Oshtran's syndrome, relapsing rheumatoid arthritis (PR), paraneoplastic cerebellar degeneration (PCD), paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg syndrome, Parsonage-Turner syndrome, pars planitis (peripheral uveitis), Streptococcus-associated paediatric autoimmune neuropsychiatric disorders (PANDAS), pelvic inflammatory disease (PID), pemphigus, pemphigus vulgaris, peripheral neuropathy, perivenous encephalomyelitis,Pernicious anemia (PA), acute pityriasis lichenoides, POEMS syndrome, polyarteritis nodosa, polyglandular syndrome type I, II, III, polymyalgia rheumatica, polymyositis, post-myocardial infarction syndrome, post-pericardiotomy syndrome, primary biliary cholangitis (PBC), primary biliary cirrhosis, primary immunodeficiency, primary sclerosing cholangitis, progesterone dermatitis, progressive inflammatory neuropathy, psoriasis, psoriatic arthritis, pure red cell aplasia (PRCA), pyoderma gangrenosum, Rasmussen encephalitis, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, relapsing polychondritis, restless legs syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, rheumatoid vasculitis, sarcoidosis, schizophrenia, schwannosis ... These include Mitt syndrome, Schnitzler syndrome, scleritis, scleroderma, serum sickness, Sjögren's syndrome, spermatozoonotic autoimmune reactions, spondyloarthropathy, stiff-person syndrome (SPS), subacute bacterial endocarditis (SBE), Sazac syndrome, Sweet's syndrome, Sydenham chorea, sympathetic ophthalmia (SO), systemic lupus erythematosus (SLE), Takayasu's arteritis, temporal arteritis / giant cell arteritis, thrombocytopenia, thrombocytopenic purpura (TTP), thyroid disease, Tolosa-Hunt syndrome (THS), transverse myelitis, type 1 diabetes, ulcerative colitis (UC), undifferentiated connective tissue disease (UCTD), undifferentiated spondyloarthritis, urticarial vasculitis, urticaria, uveitis, vasculitis, vitiligo, and Vogt-Koyanagi-Harada disease. In various embodiments, inflammatory diseases treatable by the methods and compositions of the present disclosure are treated while sparing healthy cells. ,

[0317] In various embodiments, infectious diseases that can be treated by the methods and compositions of the present disclosure include, but are not limited to, Acinetobacter infections, actinomycosis, acute flaccid myelitis (AFM), African sleeping sickness (African trypanosomiasis), AIDS (acquired immune deficiency syndrome), amebic infections, amebic dysentery, Anaplasma phagocytophilum infection, anaplasmosis, angiostrongyliasis, anisakiasis, anthrax, arboviral diseases, neuroinvasive and non-neuroinvasive, Arcanobacterium haemolyticum infection, Argentine hemorrhagic fever, ascariasis, aspergillosis, astrovirus infection, avian influenza, babesiosis, Bacillus cereus infection, bacterial infection, bacterial meningitis, bacterial pneumonia, bacterial vaginosis, Bacteroides infection, Balantidiosis, Bartonellosis, Baylisascaris infection, BK virus infection, Black sand mites disease, Blastocystosis, Bolivian hemorrhagic fever, botulism, botulism (foodborne), botulism (infants), botulism (other), botulism (wound), Brazilian hemorrhagic fever, brucellosis, bubonic plague, Burkholderia infection, Buruli ulcer, calicivirus infection (norovirus and sapovirus), California serogroup viral disease, Campylobacter, campylobacteriosis, Candida auris, clinical, candidiasis (moloniasis, thrush), capillariasis, carbapenemase-producing carbapenem-resistant enterobacteriaceae (CP-CRE), carbapenem-resistant infections (CRE / CRPA), Carrion's disease, cat scratch disease, cellulitis, Chagas' disease (trypanosomiasis), chancroid, chickenpox, chikungunya virus infection (chikungunya), chlamydia, Chlamydia trachomatis, Chlamydia pneumonia infection, cholera, chromoblastomycosis, chytridiomycosis, ciguatera, Clonorchiasis, Clostridium difficile enteritis, Clostridium difficile infection, Clostridium perfringens, Coccidioides fungal infection (Valley fever), Colorado tick fever (CTF), cold (acute viral nasopharyngitis, acute coryza), congenital syphilis, conjunctivitis, COVID-19 (coronavirus disease 2019), CP-CRE, Enterobacterspp., CP-CRE, Escherichia coli (E. coli), CP-CRE, Klebsiella spp., Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy (CJD), Creutzfeldt-Jakob disease (CJD), Crimean-Congo hemorrhagic fever (CCHF), crusted scabies, cryptococcosis, cryptosporidiosis (Crypto), cutaneous larva migrans (CLM), Cyclospora, cyclosporosis, cysticercosis, cytomegalovirus infection, dengue virus infection, dengue types 1, 2, 3, and 4 (dengue fever), dengue-like illness, Desmodesmus infection, diarrheal illness, dientamebiasis, diphtheria, diphyllobothriasis, dracunculiasis, E. coli, E. coli Infection, Shiga toxin-producing (STEC), Eastern Equine Encephalitis disease, Ebola hemorrhagic fever (Ebola), Echinococcosis, Ehrlichia chaffeensis infection, Ehrlichia ewingii infection, Ehrlichiosis, Anaplasmosis, Encephalitis, Arbovirus or parasitic infection, Helminthiasis (pinworm infection), Enterococcus infection, Enterovirus infection, D68 (EV-D68), Enterovirus infection, Non-polio (non-polio enterovirus), Epstein-Barr virus, Infectious mononucleosis (Mono), Erythema infectiosum (5th disease), Exanthema subitum (6th disease), Fascioliasis, Clostridium histolyticum, Fatal familial insomnia (FFI), 5th disease, Filariasis, Flu (seasonal), Food poisoning, Clostridium perfringens food poisoning, free-living amoeba infections, fungal infections, Fusobacterium infections, gas gangrene (clostridial myonecrosis), genital herpes, genital warts, diotrichosis, rubella, Gerstmann-Straussler-Scheinker syndrome (GSS), giardiasis, glanders, gnathostomiasis, gonorrhea, groin granuloma, groin granuloma (donovanosis), group A streptococcal infections, group A streptococcus, group B streptococcus infections, Guanarito virus, Haemophilus influenzae, type B (Hib or H-flu), Haemophilus influenzae infection, hand, foot and mouth disease (HFMD), leprosy, hantavirus infection, hantavirus pulmonary syndrome (HPS), heartland virus disease, Helicobacterpylori infection, hemolytic uremic syndrome (HUS), hemorrhagic fever with renal syndrome (HFRS), Hendra virus infection, hepatitis A (Hep A), hepatitis B (Hep B), hepatitis C (Hep C), hepatitis D (Hep D), hepatitis E (Hep E), herpes, herpes B virus, herpes simplex, shingles, chickenpox (VZV), Hib disease, histoplasmosis infection (histoplasmosis), hookworm infection, HPV (human papillomavirus), human bocavirus infection, human ehrlichiosis, human granulocytic anaplasmosis (HGA), human immunodeficiency virus / AIDS (HIV / AIDS), human metapneumovirus infection, human monocytic ehrlichiosis, human papillomavirus (HPV) infection, human parainfluenza virus infection, microtaeniasis, impetigo, influenza (flu), influenza (seasonal), invasive pneumococcal infection, isosporosis, Junin virus, Kawasaki syndrome, keratitis, Kingella kingae infection, kuru, Lassa fever, Lassa virus, Legionnaires' disease (Legionnaires' disease), leishmaniasis, leprosy Disease), Leptospirosis, Listeriosis (Listeria), Lujo Virus, Lyme Disease, Lymphatic Filariasis (Elephantiasis), Lymphocytic Choriomeningitis (LCMV), Lymphogranuloma venereum Infection (LGV), Machupo Virus, Malaria, Marburg Virus Infection, Measles, Melioidosis (Whitmore Disease), Meningitis, Bacterial Meningitis, Viral Meningitis, Meningococcal Infection, Clonorchiasis, Microsporidiosis, Middle East Respiratory Syndrome (MERS), Molluscum Contagiosum (MC), Monkeypox, Mononucleosis, Mosquito-Borne Disease, MRSA, Mumps, Typhus (Typhus), Mycetoma, Mycoplasma genitalium Infection, Mycoplasmal Pneumonia, Myiasis, Neisseria meningitidis, neonatal conjunctivitis (ophthalmia neonatorum), Nipah virus infection, nocardiosis, norovirus, onchocerciasis (river blindness), opisthorchiasis, orf virus (mouth sores), paracoccidioidomycosis (South American blastomycosis), paragonimiasis, paralytic shellfish poisoning (paralytic shellfish poisoning, ciguatera), pasteurellosis, PEP, parasitic infection, pertussis (whooping cough)cough), conjunctivitis, pneumococcal infection, pneumocystis pneumonia (PCP), pneumonia, pneumonic plague, acute poliomyelitis (polio), acute poliomyelitis, paralytic, poliovirus infection, Pontiac fever, Powassan virus disease, Prevotella infection, primary amebic meningoencephalitis (PAM), progressive multifocal leukoencephalopathy, protozoan infection, psittacosis (parrot fever), papule (smallpox, monkeypox, cowpox), rabies, raccoon roundworm, rat-bite fever, recreational water diseases Illnesses, relapsing fever, respiratory syncytial virus infection, Reye's syndrome, rhinosporidiosis, rhinovirus infection, rickettsial infection, rickettsiosis (Rocky Mountain spotted fever), Rift Valley fever (RVF), ringworm, rotavirus infection, rubella, sabi virus, salmonella, Salmonella Paratyphi infection, Salmonella Typhi infection, salmonellosis, SARS (Severe Acute Respiratory Syndrome), scabies, scarlet fever, schistosomiasis, scombroidosis, sepsis, septic shock, septicemic plague, severe acute respiratory syndrome (SARS), Shiga toxin-producing Escherichia coli, Shigella, dysentery, shingles, herpes zoster zoster), smallpox, mouth sores (orf virus), sporotrichosis, spotted fever rickettsiosis, St. Louis encephalitis virus disease, Staphylococcus aureus infection, Staphylococcus aureus infection (methicillin-resistant (MRSA)), Staphylococcus aureus food poisoning, Staphylococcus aureus infection (vancomycin-intermediately resistant (VISA)), streptococcal pharyngitis, hemolytic streptococcal infection, group A (invasive) (Strep A (invasive), hemolytic streptococcal infection, group B (Strep-B), Staphylococcus aureus toxic shock syndrome, strongyloidiasis, subacute sclerosing panencephalitis, syphilis, taeniasis, tetanus infection, tick-borne diseases, tinea barbae, tinea capitis, tinea corporis, tinea cruris, tinea manubriata, tinea nigricans, tinea pedis, tinea unguium, tinea versicolor, toxic shock syndrome, toxocariasis (ocular larva migrans (OLM)), toxocariasis (visceral larva migrans (VLM)), toxoplasmosis, trachoma, trichinosis, trichomoniasis, trichinosis infection (trichinosis), trichuriasis (whipworm infection), tuberculosis (TB), tularemia (rabbit fever), typhoid fever, group D, typhoid, typhoid fever, Ureaplasmaurealyticum infection, vaginitis, Valley fever, variant Creutzfeldt-Jakob disease (vCJD, nvCJD), chickenpox (varicella), Venezuelan equine encephalitis virus disease, Venezuelan hemorrhagic fever, Vibrio cholerae (cholera), Vibrio parahaemolyticus, Vibrio vulnificus infection, vibriosis, viral infection, viral hemorrhagic fever, viral hemorrhagic fever (Ebola, Lassa, Marburg), viral hemorrhagic fever (VHF), viral pneumonia, West Nile virus disease, Western equine encephalitis virus disease, white sand cripple disease (ringworm), whooping cough, yellow fever, Yersinia In various embodiments, infectious diseases treatable by the methods and compositions of the present disclosure are treated while sparing healthy cells.

[0318] In one embodiment, the invention is a method of activating at least one γδ T cell in a subject, the method comprising administering an anti-Vδ1 antibody or fragment thereof as defined herein.

[0319] In one embodiment, the invention is a method for causing or increasing proliferation of γδ T cells in a subject, the method comprising administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0320] In one embodiment, the invention is a method for causing or increasing the expansion of γδ T cells in a subject, the method comprising administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0321] In one embodiment, the invention is a method for causing or increasing γδ T cell degranulation in a subject, the method comprising administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0322] In one embodiment, the invention is a method of causing or increasing γδ T cell killing activity in a subject, the method comprising the step of administering to the subject an anti-V51 antibody or fragment thereof as defined herein. In one embodiment, the invention is a method of causing or increasing γδ T cell killing activity in a subject, while sparing healthy cells, the method comprising the step of administering to the subject an anti-V51 antibody or fragment thereof as defined herein.

[0323] In one embodiment, the invention is a method of causing or increasing γδ T cytotoxicity in a subject, the method comprising administering to the subject an anti-V51 antibody or fragment thereof as defined herein. In one embodiment, the invention is a method of causing or increasing γδ T cytotoxicity in a subject, while sparing healthy cells, the method comprising administering to the subject an anti-V51 antibody or fragment thereof as defined herein.

[0324] In one embodiment, the invention is a method of causing or increasing γδ T cell recruitment in a subject, the method comprising administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0325] In one embodiment, the invention is a method for increasing survival of γδ T cells in a subject, the method comprising administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0326] In one embodiment, the invention is a method for increasing resistance to depletion of γδ T cells in a subject, the method comprising the step of administering to the subject an anti-Vδ1 antibody or fragment thereof as defined herein.

[0327] According to a further aspect of the present invention there is provided a method of stimulating an immune response in a subject, the method comprising administering to the subject an anti-V51 antibody or fragment thereof in an amount effective to stimulate the immune response.

[0328] Uses of antibodies or fragments thereof According to a further aspect of the present invention, there is provided a use of an anti-V51 antibody or fragment thereof as described herein for studying antigen recognition, activation, signaling or function of γδ T cells (particularly V51 T cells). As described herein, the antibodies have been shown to be active in assays that can be used to examine γδ T cell function. Such antibodies may also be useful for inducing proliferation of γδ T cells and therefore may be used in methods of expanding γδ T cells (e.g., V51 T cells).

[0329] Antibodies that bind to the V51 chain can be used to detect γδ T cells. For example, the antibodies can be labeled with a detectable label or reporter molecule, or can be used as a capture ligand to selectively detect and / or isolate V51 T cells in a sample. Labeled antibodies find use in many methods known in the art, such as immunohistochemistry and ELISA.

[0330] The detectable label or reporter molecule may be 3 H, 14 C. 32 P, 35 S or 125 The fluorescent label may be a radioisotope such as I, a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate or rhodamine, or an enzyme such as alkaline phosphatase, β-galactosidase, horseradish peroxidase, or luciferase. Fluorescent labels applied to antibodies of the invention can then be used in fluorescence-activated cell sorting (FACS) methods.

[0331] Thus, in various embodiments, the present invention includes in vivo methods of modulating γδ T cells, methods of binding to γδ T cells, methods of targeting γδ T cells, methods of activating γδ T cells, methods of proliferating γδ T cells, methods of expanding γδ T cells, methods of detecting γδ T cells, methods of inducing γδ T cell degranulation, methods of inducing γδ T cell killing activity, methods of selecting an antibody or fragment thereof, and methods comprising the step of administering an anti-γδ antibody or fragment thereof to a subject as described herein.

[0332] section A series of clauses defining the invention and its preferred embodiments are set forth below.

[0333] 1. A method for treating cancer, an infectious disease or an inflammatory disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-vδ1 antibody or a fragment thereof.

[0334] 2. The anti-Vδ1 antibody or fragment thereof CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26 to 37 and A1 to A12 (in Table 3), and / or 2. The method of claim 1, comprising one or more of the CDR1s comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38-61.

[0335] 3. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2-13, such as SEQ ID NOs: 8, 9, 10, or 11.

[0336] 4. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26-37, such as SEQ ID NOs: 32, 33, 34, or 35.

[0337] 5. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38-49, such as SEQ ID NOs: 44, 45, 46, or 47.

[0338] 6. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 8, a CDR2 comprising the sequence of SEQ ID NO: 32, and a CDR1 comprising the sequence of SEQ ID NO: 44.

[0339] 7. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 9, a CDR2 comprising the sequence of SEQ ID NO: 33, and a CDR1 comprising the sequence of SEQ ID NO: 45.

[0340] 8. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 10, a CDR2 comprising the sequence of SEQ ID NO: 34, and a CDR1 comprising the sequence of SEQ ID NO: 46.

[0341] 9. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 11, a CDR2 comprising the sequence of SEQ ID NO: 35, and a CDR1 comprising the sequence of SEQ ID NO: 47.

[0342] 10. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14-25, such as SEQ ID NOs: 20, 21, 22, or 23.

[0343] 11. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1-A12, e.g., SEQ ID NO: A7, A8, A9, or A10.

[0344] 12. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 50-61, such as SEQ ID NOs: 56, 57, 58, or 59.

[0345] 13. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 20, a CDR2 comprising the sequence of sequence A7, and a CDR1 comprising the sequence of SEQ ID NO: 56.

[0346] 14. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 21, a CDR2 comprising the sequence of sequence A8, and a CDR1 comprising the sequence of SEQ ID NO: 57.

[0347] 15. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 22, a CDR2 comprising the sequence of sequence A9, and a CDR1 comprising the sequence of SEQ ID NO: 58.

[0348] 16. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 23, a CDR2 comprising the sequence of sequence A10, and a CDR1 comprising the sequence of SEQ ID NO: 59.

[0349] 17. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 6, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 13.

[0350] 18. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 7 and the CDR1, CDR2 and CDR3 sequences defined in clause 14.

[0351] 19. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 8 and the CDR1, CDR2 and CDR3 sequences defined in clause 15.

[0352] 20. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises the CDR1, CDR2 and CDR3 sequences defined in clause 9 and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 16.

[0353] 21. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62-85.

[0354] 22. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62-73.

[0355] 23. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62, 63, 64, 68, 69, 70 or 71.

[0356] 24. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74-85.

[0357] 25. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74, 75, 76, 80, 81, 82 or 83.

[0358] 26. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 62 and a VL region comprising the amino acid sequence of SEQ ID NO: 74.

[0359] 27. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 63 and a VL region comprising the amino acid sequence of SEQ ID NO: 75.

[0360] 28. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 64 and a VL region comprising the amino acid sequence of SEQ ID NO: 76.

[0361] 29. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 68 and a VL region comprising the amino acid sequence of SEQ ID NO: 80.

[0362] 30. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 69 and a VL region comprising the amino acid sequence of SEQ ID NO: 81.

[0363] 31. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 70 and a VL region comprising the amino acid sequence of SEQ ID NO: 82.

[0364] 32. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 71 and a VL region comprising the amino acid sequence of SEQ ID NO: 83.

[0365] 33. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises a VH region and a VL region, and the VH region and the VL region are linked by a linker, such as a polypeptide linker.

[0366] 34. The method of clause 33, wherein the linker comprises a (Gly4Ser)n format, where n=1-8.

[0367] 35. The method of clause 33 or clause 34, wherein the linker comprises a [(Gly4Ser)n(Gly3AlaSer)m]p linker, where n, m and p=1-8.

[0368] 36. The method of clause 33 or clause 35, wherein the linker comprises SEQ ID NO: 98.

[0369] 37. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 86-97.

[0370] 38. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises the amino acid sequence of any one of SEQ ID NOs: 86-97.

[0371] 39. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises SEQ ID NO: 86.

[0372] 40. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises SEQ ID NO: 87.

[0373] 41. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises SEQ ID NO: 88.

[0374] 42. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises SEQ ID NO: 92.

[0375] 43. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises SEQ ID NO: 93.

[0376] 44. The method of clause 1, wherein the anti-V51 antibody or fragment thereof comprises SEQ ID NO: 94.

[0377] 45. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises SEQ ID NO: 95.

[0378] 46. ​​The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof binds to or competes with the same or essentially the same epitope as an antibody or fragment thereof defined in any one of clauses 1 to 45.

[0379] 47. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof comprises and binds an epitope of the variable delta 1 (Vδ1) chain of the gamma delta T cell receptor (TCR) comprising one or more amino acid residues within amino acids 1 to 90 of SEQ ID NO: 1.

[0380] 48. The method of clause 47, wherein the epitope comprises at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO:1.

[0381] 49. The method of clause 47 or 48, wherein the epitope comprises one or more amino acid residues within amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:1.

[0382] 50. The method of any one of clauses 47-49, wherein the epitope consists of one or more amino acid residues within amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:1.

[0383] 51. The method of any one of clauses 47-50, wherein the epitope comprises one or more amino acid residues within amino acid regions 5-20 and 62-77 of SEQ ID NO:1.

[0384] 52. The method of any one of clauses 47-51, wherein the epitope comprises one or more amino acid residues within amino acid region 50-64 of SEQ ID NO:1.

[0385] 53. The method of any one of clauses 47-52, wherein the epitope comprises one or more amino acid residues within amino acid regions 37-53 and 59-77 of SEQ ID NO:1.

[0386] 54. The method of any one of clauses 47-53, wherein the epitope is an activating epitope of γδ T cells.

[0387] 55. The method of clause 54, wherein binding of the activating epitope (i) downregulates γδ TCR, (ii) activates γδ T cell degranulation, and / or (iii) activates γδ T cell killing.

[0388] 56. The method of any one of clauses 47-55, wherein the antibody or fragment thereof binds only to an epitope in the V region of the Vδ1 chain of a γδ TCR.

[0389] 57. The method of any one of clauses 47-56, wherein the antibody or fragment thereof does not bind to an epitope found in the CDR3 of the Vδ1 chain of the γδ TCR.

[0390] 58. The method of clause 57, wherein the antibody or fragment thereof does not bind to an epitope within amino acid region 91-105 (CDR3) of SEQ ID NO:1.

[0391] 59. The method of any one of clauses 1 to 58, wherein the antibody or fragment thereof binds to the variable delta 1 (Vδ1) chain of the gamma delta T cell receptor (TCR) with a binding affinity (KD) of less than 1.5 x 107 M, as measured by surface plasmon resonance.

[0392] 60. The method of clause 59, wherein the antibody or fragment thereof exhibits a KD of less than 1.3x10-7M or less, such as less than 1.0x10-7M, in particular less than 5.0x10-8M.

[0393] 61. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for down-regulation of γδ TCR upon binding of less than 0.5 μg / ml.

[0394] 62. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for down-regulation of γδ TCR upon binding of less than 0.06 μg / ml.

[0395] 63. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for γδ T cell degranulation upon binding of less than 0.05 μg / ml.

[0396] 64. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for γδ T cell degranulation upon binding of less than 0.005 μg / ml, such as less than 0.002 μg / ml.

[0397] 65. The method of clause 63 or clause 64, wherein the γδ T cell degranulation EC50 value is measured by detecting CD107a expression.

[0398] 66. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for γδ T cell killing upon binding of less than 0.5 μg / ml.

[0399] 67. The method of clause 1, wherein the antibody or fragment thereof has an EC50 value for γδ T cell killing upon binding of less than 0.055 μg / ml, such as less than 0.020 μg / ml.

[0400] 68. The method of any one of clauses 61 to 67, wherein the EC50 value is measured using flow cytometry.

[0401] 69. The method of clause 1, wherein the antibody or fragment thereof defined in any one of clauses 1 to 68 is an scFv, Fab, Fab', F(ab')2, Fv, a variable domain (e.g., VH or VL), a diabody, a minibody, or a full-length antibody.

[0402] 70. The method of clause 69, wherein the antibody or fragment thereof is an scFv, or a full-length antibody, such as an IgG1.

[0403] 71. The method of clause 70, wherein the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 111-122.

[0404] 72. The method of clause 70 or clause 71, wherein the antibody or fragment thereof comprises the amino acid sequence of any one of SEQ ID NOs: 111-122.

[0405] 73. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 111.

[0406] 74. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 112.

[0407] 75. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 116.

[0408] 76. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 117.

[0409] 77. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 118.

[0410] 78. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 119.

[0411] 79. The method of clause 71 or clause 72, wherein the antibody or fragment thereof comprises SEQ ID NO: 120.

[0412] 80. The method of any one of clause 1 or clause 79, wherein the antibody or fragment thereof is human.

[0413] 81. The method of clause 1, wherein the anti-Vδ1 antibody or fragment thereof has an EC50 value for down-regulation of γδ TCR upon binding of less than 0.5 μg / ml, an EC50 value for γδ T cell degranulation upon binding of less than 0.05 μg / ml, and / or an EC50 value for γδ T cell killing upon binding of less than 0.5 μg / ml.

[0414] 82. A method of modulating an immune response in a subject in need thereof, comprising administering to the subject an anti-vδ1 antibody or fragment thereof as defined in any one of clauses 1 to 81.

[0415] 83. The method of clause 82, wherein the subject has cancer, an infectious disease, or an inflammatory disease.

[0416] 84. The method of clause 82, wherein modulating the immune response in the subject comprises at least one selected from the group consisting of activating γδ T cells, causing or increasing γδ T cell proliferation, causing or increasing γδ T cell expansion, causing or increasing γδ T cell degranulation, causing or increasing γδ T cell killing activity, causing or increasing γδ T cell toxicity, causing or increasing γδ T cell recruitment, increasing γδ T cell survival, and increasing resistance to γδ T cell depletion.

[0417] 85. The method of any one of clauses 1-84, wherein diseased cells are killed while healthy cells are spared.

[0418] 86. An isolated multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first target antigen of the at least two target antigens is Vδ1; CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26 to 37 and A1 to A12 (in Table 3), and / or An isolated multispecific antibody or a fragment thereof, comprising one or more CDR1s comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 61.

[0419] 87. The isolated multispecific antibody or fragment thereof defined in clause 86, comprising a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2-13, such as SEQ ID NOs: 8, 9, 10, or 11.

[0420] 88. The isolated multispecific antibody or fragment thereof as defined in clause 86 or clause 87, comprising a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26-37, such as SEQ ID NOs: 32, 33, 34, or 35.

[0421] 89. The isolated multispecific antibody or fragment thereof defined in any one of clauses 86-88, comprising a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38-49, such as SEQ ID NOs: 44, 45, 46, or 47.

[0422] 90. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 89, comprising a VH region comprising: a CDR3 comprising the sequence of SEQ ID NO: 8; a CDR2 comprising the sequence of SEQ ID NO: 32; and a CDR1 comprising the sequence of SEQ ID NO: 44.

[0423] 91. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 89, comprising a VH region comprising: a CDR3 comprising the sequence of SEQ ID NO: 9; a CDR2 comprising the sequence of SEQ ID NO: 33; and a CDR1 comprising the sequence of SEQ ID NO: 45.

[0424] 92. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 89, comprising a VH region comprising: a CDR3 comprising the sequence of SEQ ID NO: 10; a CDR2 comprising the sequence of SEQ ID NO: 34; and a CDR1 comprising the sequence of SEQ ID NO: 46.

[0425] 93. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 89, comprising a VH region comprising: a CDR3 comprising the sequence of SEQ ID NO: 11; a CDR2 comprising the sequence of SEQ ID NO: 35; and a CDR1 comprising the sequence of SEQ ID NO: 47.

[0426] 94. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86-93, comprising a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14-25, such as SEQ ID NOs: 20, 21, 22, or 23.

[0427] 95. The isolated multispecific antibody or fragment thereof defined in any one of clauses 86-94, comprising a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1-A12, e.g., SEQ ID NO: A7, A8, A9, or A10.

[0428] 96. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86-95, comprising a VL region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 50-61, such as SEQ ID NOs: 56, 57, 58, or 59.

[0429] 97. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 95, comprising a VL region comprising: a CDR3 comprising the sequence of SEQ ID NO: 20; a CDR2 comprising the sequence of sequence A7; and a CDR1 comprising the sequence of SEQ ID NO: 56.

[0430] 98. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 95, comprising a VL region comprising: a CDR3 comprising the sequence of SEQ ID NO: 21; a CDR2 comprising the sequence of sequence A8; and a CDR1 comprising the sequence of SEQ ID NO: 57.

[0431] 99. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 95, comprising a VL region comprising: a CDR3 comprising the sequence of SEQ ID NO: 22; a CDR2 comprising the sequence of sequence A9; and a CDR1 comprising the sequence of SEQ ID NO: 58.

[0432] 100. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 95, comprising a VL region comprising: a CDR3 comprising the sequence of SEQ ID NO: 23; a CDR2 comprising the sequence of sequence A10; and a CDR1 comprising the sequence of SEQ ID NO: 59.

[0433] 101. An isolated multispecific antibody or fragment thereof, comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in paragraph 90, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in paragraph 97.

[0434] 102. An isolated multispecific antibody or fragment thereof, comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in paragraph 91, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in paragraph 98.

[0435] 103. An isolated multispecific antibody or fragment thereof, comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 92, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 100.

[0436] 104. An isolated multispecific antibody or fragment thereof, comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 93, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 100.

[0437] 105. An isolated multispecific antibody or fragment thereof, comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62 to 85.

[0438] 106. The isolated multispecific antibody or fragment thereof defined in clause 49, comprising a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62-73.

[0439] 107. The isolated multispecific antibody or fragment thereof as defined in clause 50, wherein the VH region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62, 63, 64, 68, 69, 70, or 71.

[0440] 108. The isolated multispecific antibody or fragment thereof defined in any one of clauses 105-107, comprising a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74-85.

[0441] 109. The isolated multispecific antibody or fragment thereof defined in clause 108, wherein the VL region comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74, 75, 76, 80, 81, 82, or 83.

[0442] 110. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 62 and a VL region comprising the amino acid sequence of SEQ ID NO: 74.

[0443] 111. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 63, and a VL region comprising the amino acid sequence of SEQ ID NO: 75.

[0444] 112. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 64, and a VL region comprising the amino acid sequence of SEQ ID NO: 76.

[0445] 113. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 68, and a VL region comprising the amino acid sequence of SEQ ID NO: 80.

[0446] 114. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 69 and a VL region comprising the amino acid sequence of SEQ ID NO: 81.

[0447] 115. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 70, and a VL region comprising the amino acid sequence of SEQ ID NO: 82.

[0448] 116. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105 to 109, comprising a VH region comprising the amino acid sequence of SEQ ID NO: 71 and a VL region comprising the amino acid sequence of SEQ ID NO: 83.

[0449] 117. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 105-116, wherein the VH region and the VL region are linked by a linker, such as a polypeptide linker.

[0450] 118. The isolated multispecific antibody or fragment thereof defined in paragraph 117, wherein the linker comprises a (Gly4Ser)n format, where n=1-8.

[0451] 119. The isolated multispecific antibody or fragment thereof defined in clause 117 or clause 118, wherein the linker comprises a [(Gly4Ser)n(Gly3AlaSer)m]p linker, and n, m and p=1-8.

[0452] 120. The isolated multispecific antibody or fragment thereof defined in any one of clauses 117-118, wherein the linker comprises SEQ ID NO: 98.

[0453] 121. The isolated multispecific antibody or fragment thereof defined in paragraph 120, wherein the linker consists of SEQ ID NO: 98.

[0454] 122. An isolated multispecific antibody or fragment thereof, comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 86 to 97.

[0455] 123. The isolated multispecific antibody or fragment thereof as defined in clause 122, comprising the amino acid sequence of any one of SEQ ID NOs: 86-97.

[0456] 124. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 86.

[0457] 125. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 87.

[0458] 126. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 88.

[0459] 127. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 92.

[0460] 128. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 93.

[0461] 129. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 94.

[0462] 130. The isolated multispecific antibody or fragment thereof as defined in clause 94 or clause 122, comprising SEQ ID NO: 95.

[0463] 131. An isolated multispecific antibody or fragment thereof that binds to or competes with the same or essentially the same Vδ1 epitope as an antibody or fragment thereof defined in any one of paragraphs 86 to 130.

[0464] 132. An isolated human anti-TCR delta variable 1 multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first target antigen of the at least two target antigens is Vδ1, and wherein the multispecific antibody or fragment thereof that binds to an epitope of Vδ1 comprises one or more amino acid residues within amino acids 1 to 90 of SEQ ID NO: 1.

[0465] 133. The human isolated multispecific antibody or fragment thereof defined in clause 132, wherein the epitope comprises at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO:1.

[0466] 134. The human isolated multispecific antibody or fragment thereof defined in clause 132 or clause 133, wherein the epitope comprises one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:1.

[0467] 135. The human isolated multispecific antibody or fragment thereof defined in clause 134, wherein the epitope consists of one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:1.

[0468] 136. The human isolated multispecific antibody or fragment thereof defined in any one of clauses 132-135, wherein the epitope comprises one or more amino acid residues within amino acid regions 5-20 and 62-77 of SEQ ID NO:1.

[0469] 137. The human isolated multispecific antibody or fragment thereof defined in any one of clauses 132-135, wherein the epitope comprises one or more amino acid residues within amino acid region 50-64 of SEQ ID NO:1.

[0470] 138. The human isolated multispecific antibody or fragment thereof defined in any one of clauses 132-135, wherein the epitope comprises one or more amino acid residues within amino acid regions 37-53 and 59-77 of SEQ ID NO:1.

[0471] 139. The human isolated multispecific antibody or fragment thereof defined in any one of clauses 132-138, wherein the epitope is an activating epitope of γδ T cells.

[0472] 140. A human isolated multispecific antibody or fragment thereof as defined in clause 139, wherein binding of the activating epitope (i) downregulates γδ TCR, (ii) activates γδ T cell degranulation, and / or (iii) activates γδ T cell killing.

[0473] 141. A human isolated multispecific antibody or fragment thereof as defined in any one of clauses 132-140, which binds only to an epitope in the V region of the Vδ1 chain of a γδ TCR.

[0474] 142. A human isolated multispecific antibody or fragment thereof as defined in any one of clauses 132-141, which does not bind to an epitope found in the CDR3 of the Vδ1 chain of a γδ TCR.

[0475] 143. The human isolated multispecific antibody or fragment thereof defined in clause 142, wherein the antibody or fragment does not bind to an epitope within amino acid region 91-105 (CDR3) of SEQ ID NO:1.

[0476] 144. An isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 143, which binds to the variable delta 1 (Vδ1) chain of a gamma delta T cell receptor (TCR) with a binding affinity (KD) of less than 1.5 x 107 M as measured by surface plasmon resonance.

[0477] 145. The isolated multispecific antibody or fragment thereof as defined in clause 144, wherein the KD is less than 1.3x10-7M, such as less than 1.0x10-7M, in particular less than 5.0x10-8M.

[0478] 146. An isolated multispecific antibody or fragment thereof, which has an EC50 value for down-regulation of γδ TCR upon binding of less than 0.5 μg / ml.

[0479] 147. An isolated multispecific antibody or fragment thereof, which has an EC50 value for down-regulation of γδ TCR upon binding of less than 0.06 μg / ml.

[0480] 148. An isolated multispecific antibody or fragment thereof, which has an EC50 value for γδ T cell degranulation upon binding of less than 0.05 μg / ml.

[0481] 149. An isolated multispecific antibody or fragment thereof, which has an EC50 value for γδ T cell degranulation upon binding of less than 0.005 μg / ml, for example less than 0.002 μg / ml.

[0482] 150. The isolated multispecific antibody or fragment thereof defined in clause 148 or clause 149, wherein the gamma delta T cell degranulation EC50 value is measured by detecting CD107a expression.

[0483] 151. An isolated multispecific antibody or fragment thereof, which has an EC50 value for γδ T cell killing upon binding of less than 0.5 μg / ml.

[0484] 152. An isolated multispecific antibody or fragment thereof, which has an EC50 value for γδ T cell killing upon binding of less than 0.055 μg / ml, such as less than 0.020 μg / ml.

[0485] 153. The isolated multispecific antibody or fragment thereof defined in any one of clauses 146 to 152, wherein the EC50 value is measured using flow cytometry.

[0486] 154. An isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 153, which is an scFv, Fab, Fab', F(ab')2, Fv, a variable domain (e.g., VH or VL), a diabody, a minibody, or a full-length antibody.

[0487] 155. An isolated multispecific antibody or fragment thereof as defined in clause 154, which is an scFv, or a full-length antibody, e.g., an IgG1.

[0488] 156. The isolated multispecific antibody defined in clause 155, comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 111-122.

[0489] 157. The isolated multispecific antibody as defined in clause 155 or clause 156, comprising the amino acid sequence of any one of SEQ ID NOs: 111-122.

[0490] 158. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 111.

[0491] 159. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 112.

[0492] 160. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 116.

[0493] 161. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 117.

[0494] 162. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 118.

[0495] 163. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 119.

[0496] 164. The isolated multispecific antibody defined in clause 155 or clause 156, comprising SEQ ID NO: 120.

[0497] 165. The isolated multispecific antibody or fragment thereof defined in any one of clauses 86 to 164, which is human.

[0498] 166. The second target antigen of the at least two target antigens is selected from the group consisting of CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CD13, CD14, CD15, CD16, CD16a, CD16b, CD17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, and CD3 2A, CD32B, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60a, CD60b, CD60c, CD61, CD62E, CD62L, CD62P, CD63, CD64a , CD65, CD65s, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD75s, CD77, CD79A, CD79B, CD80, CD81, CD82 , CD83, CD84, CD85A, CD85B, CD85C, CD85D, CD85F, CD85G, CD85H, CD85I, CD85J, CD85K, CD85M, CD86, CD87, CD88, CD89, CD90, CD91, CD92, CD93, CD94, CD9 5, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107, CD107a, CD107b, CD108, CD109, CD110, CD111, CD112, CD113, CD114 , CD115, CD116, CD117, CD118, CD119, CD120, CD120a, CD120b, CD121a, CD121b, CD122, CD123, CD124, CD125, CD126, CD127, CD129, CD130, CD131, CD132,CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140A、CD140B、CD141、CD142、CD143、CD144、CDw145、CD146、CD147、CD148、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD157、CD158、CD158A、CD158B1、CD158B2、CD158C、CD158D、CD158E1、CD158E2、CD158F1、CD158F2、CD158G、CD158H、CD158I、CD158J、CD158K、CD159a、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CDw198、CDw199、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218a、CD218b、CD219、CD220、CD221、CD222、CD223、CD224、CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240CE、CD240D、CD241、CD242、CD243、CD244、CD245

[17] 、CD246、CD247、CD248、CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261, CD262, CD263, CD264, CD265, CD266, CD267, CD268, CD269, CD270, CD271, CD272, CD273, CD274, CD27 5, CD276, CD277, CD278, CD279, CD280, CD281, CD282, CD283, CD284, CD285, CD286, CD287, CD288, CD289, CD2 90, CD291, CD292, CDw293, CD294, CD295, CD296, CD297, CD298, CD299, CD300A, CD300C, CD301, CD302, CD303 , CD304, CD305, CD306, CD307, CD307a, CD307b, CD307c, CD307d, CD307e, CD308, CD309, CD310, CD311, CD312 , CD313, CD314, CD315, CD316, CD317, CD318, CD319, CD320, CD321, CD322, CD323, CD324, CD325, CD326, CD3 27, CD328, CD329, CD330, CD331, CD332, CD333, CD334, CD335, CD336, CD337, CD338, CD339, CD340, CD344, CD 349, CD351, CD352, CD353, CD354, CD355, CD357, CD358, CD360, CD361, CD362, CD363, CD364, CD365, CD366, CD367, CD368, CD369, CD370, and CD371.

[0499] 167. The second target antigen of the at least two target antigens is AFP, AKAP-4, ALK, alpha-fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCAA, Bcr-abl, beta-catenin, beta-HCG, beta-human chorionic gonadotropin, BORIS, BTAA, CA 125, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, or CAM. 17.1, CAM43, carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33 / IL3Ra, CD68 / P1, CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG4), c-Met, CO-029, CSPG4, cyclin B1, cyclophilin C-related protein, CYP1B1, E2A-PRL, EGFR, EGFRvIII, ELF2M, EpCAM, EphA2, ephrin B2, Epstein-Barr virus antigen EBVA, ERG (TMPRSS2ETS fusion gene), ETV6-AML, FAP, FGF-5, Fos-related antigen 1, fucosyl GM1, G250, Ga733 / EpCAM, GAGE-1, GAGE-2, GD2, GD3, glioma-associated antigen, GloboH, glycolipid F77, GM3, GP 100, GP 100 (Pmel 17), H4-RET, HER-2 / neu, HER-2 / Neu / ErbB-2, high molecular weight melanoma-associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin growth factor (IGF)-I, intestinal carboxylesterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, legumain, LMP2, M344, MA-50, Mac-2 binding protein, MAD-CT-1, MAGE, MAGE A1, MAGE A3, MAGE-1, MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1, MART-1 / MelanA, M-CSF, melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin, MG7-Ag, ML-IAP, MN-CAIX, MOV18, MUC1, Mum-1, hsp70-2, MYCN, MYL-RAR, NA17, NB / 70K, neuron-glial antigen 2 (NG2), neutrophil elastase, nm-23H1, NuMa, NY-BR-1, NY-CO-1, NY-ESO, NY-ESO-1, NY-ESO-1, OY-TES1, p15, p16, p180erbB3, p185erbB2, p53, p53 mutant, Page4, PAX3, PAX5, PDGFR-based TA, PLAC1, polysialic acid, prostate carcinoma tumor antigen-1 (PCTA-1), prostate-specific antigen, prostatic acid phosphatase (PAP), proteinase 3 (PR1), PSA, PSCA, PSMA, RAGE-1, Ras, Ras-mutants, RCAS1, RGS5, RhoC, ROR1, RU1, RU2 (AS), SART3, SDCCAG16, sLe(a), sperm protein 17, SSX2, STn, survivin, TA-90, TAAL6, a 166. The isolated polyspecific antibody or fragment thereof defined in any one of clauses 86 to 165, wherein the antibody or fragment is selected from the group consisting of TAG-72, telomerase, thyroglobulin, Tie 2, TIGIT, TLP, Tn, TPS, TRP-1, TRP-2, TRP-2, TSP-180, tyrosinase, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72.

[0500] 168. Multispecific antibody formats include CrossMab, DAF (two-in-one), DAF (four-in-one), DutaMab, DT-IgG, knob-in-hole (KIH), knob-in-hole (common light chain), charge pair, Fab arm exchange, SEED body, triomab, LUZ-Y, Fcab, κλ-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L,H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG (four-in-one), nanobody, Nanoby-HAS, BiTE, diabody, DART, TandAb, sc diabody, sc diabody-CH3, diabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 166. The isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 165, wherein the antibody or fragment is selected from the group consisting of KIH, Fab-scFv, scFV-CH-CL-scFv, F(ab')2, F(ab;)2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, sc diabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock-and-lock, ImmTAC, HSA body, sc diabody-HAS, tandem scFv-toxin, IgG-IgG, ov-X-Body, duobody, mab2, and scFv1-PEG-scFv2.

[0501] 169. A polynucleotide sequence encoding a multispecific antibody or fragment thereof as defined in any one of paragraphs 86 to 168.

[0502] 170. A polynucleotide sequence encoding a multispecific antibody or a fragment thereof comprising a sequence having at least 70% sequence identity to SEQ ID NOs: 99 to 110.

[0503] 171. Polynucleotide sequence encoding a multispecific antibody or a fragment thereof comprising the sequence of SEQ ID NO: 99 to 110.

[0504] 172. An expression vector encoding a multispecific antibody or fragment thereof comprising a polynucleotide sequence as defined in any one of paragraphs 169 to 171.

[0505] 173. An expression vector encoding a multispecific antibody or a fragment thereof comprising a VH region of SEQ ID NO: 99 to 110.

[0506] 174. An expression vector encoding a multispecific antibody or a fragment thereof comprising a VL region of SEQ ID NO: 99 to 110.

[0507] 175. An expression vector encoding a multispecific antibody or fragment thereof comprising the VH region of clause 173 and the VL region of clause 174.

[0508] 176. A cell comprising a polynucleotide sequence encoding a multispecific antibody or fragment thereof as defined in any one of paragraphs 169-171 or an expression vector as defined in any one of paragraphs 172-175.

[0509] 177. A cell comprising a first expression vector encoding a multispecific antibody or fragment thereof as defined in paragraph 173, and a second expression vector encoding a multispecific antibody or fragment thereof as defined in paragraph 174.

[0510] 178. A cell comprising an expression vector encoding a multispecific antibody or fragment thereof as defined in paragraph 175.

[0511] 179. A cell as defined in any one of clauses 176-178, wherein the polynucleotide or expression vector encoding the multispecific antibody or fragment thereof encodes a membrane anchor or transmembrane domain fused to the antibody or fragment thereof, and the antibody or fragment thereof is displayed on the extracellular surface of the cell.

[0512] 180. A composition comprising a multispecific antibody or fragment thereof as defined in any one of paragraphs 86 to 168.

[0513] 181. A pharmaceutical composition comprising a multispecific antibody or fragment thereof as defined in any one of paragraphs 86-168 together with a pharmaceutically acceptable diluent or carrier.

[0514] 182. An isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 168, or a pharmaceutical composition as defined in clause 125, for use as a medicament.

[0515] 183. An isolated multispecific antibody or fragment thereof, or pharmaceutical composition as defined in clause 182, for use in treating cancer, an infectious disease, or an inflammatory disease.

[0516] 184. A method of treating cancer, an infectious disease or an inflammatory disease in a subject in need thereof, comprising administering a therapeutically effective amount of an isolated multispecific antibody or fragment thereof as defined in any one of clauses 86 to 168, or a pharmaceutical composition as defined in clause 125.

[0517] 184. An isolated antigen comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 123, for use in generating a multispecific antibody or fragment thereof.

[0518] 185. A method for producing a multispecific antibody or fragment thereof, comprising: (i) designing a series of antigens comprising a TCR delta variable 1 (TRDV1) amino acid sequence, wherein the CDR3 sequence of TRDV1 is the same for all antigens in the series; (ii) exposing the first antigen designed in step (i) to an antibody library; and (iii) isolating antibodies or fragments thereof that bind to the antigen. (iv) exposing the isolated antibody or fragment thereof to the second antigen designed in step (i); (v) isolating an antibody or fragment thereof that binds to both the first and second antigens.

[0519] 186. The method defined in clause 185, further comprising exposing the isolated antibodies or fragments thereof to a second set of antigens comprising a gamma delta TCR with a different delta variable chain, e.g., TCR delta variable 2 (TRDV2) or TCR delta variable 3 (TRDV3), and then deselecting antibodies or fragments thereof that also bind to the second set of antigens.

[0520] 187. The method defined in clause 185 or clause 186, wherein the first and / or second set of antigens are presented as leucine zipper and / or Fc fusions.

[0521] 188. The method defined in any one of clauses 185 to 187, wherein the set of antigens is in heterodimeric and / or homodimeric format.

[0522] 189. An antibody obtained by a method defined in any one of paragraphs 185 to 188.

[0523] 190. An anti-vδ1 antibody or fragment thereof for use in a method for treating cancer, an infectious disease or an inflammatory disease.

[0524] 191. An anti-Vδ1 antibody or a fragment thereof CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26 to 37 and A1 to A12 (in Table 3), and / or 191. The anti-vδ1 antibody or fragment thereof of clause 190, comprising one or more of the CDR1s comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38-61.

[0525] 192. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 2-13, such as SEQ ID NOs: 8, 9, 10, or 11.

[0526] 193. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 26-37, such as SEQ ID NOs: 32, 33, 34, or 35.

[0527] 194. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38-49, such as SEQ ID NOs: 44, 45, 46, or 47.

[0528] 195. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 8, a CDR2 comprising the sequence of SEQ ID NO: 32, and a CDR1 comprising the sequence of SEQ ID NO: 44.

[0529] 196. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 9, a CDR2 comprising the sequence of SEQ ID NO: 33, and a CDR1 comprising the sequence of SEQ ID NO: 45.

[0530] 197. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 10, a CDR2 comprising the sequence of SEQ ID NO: 34, and a CDR1 comprising the sequence of SEQ ID NO: 46.

[0531] 198. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 11, a CDR2 comprising the sequence of SEQ ID NO: 35, and a CDR1 comprising the sequence of SEQ ID NO: 47.

[0532] 199. The anti-vδ1 antibody or fragment thereof of clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 14-25, such as SEQ ID NOs: 20, 21, 22, or 23.

[0533] 200. The anti-vδ1 antibody or fragment thereof of clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR2 comprising a sequence having at least 80% sequence identity to any one of sequences A1 to A12, e.g., SEQ ID NO: A7, A8, A9, or A10.

[0534] 201. The anti-vδ1 antibody or fragment thereof of clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 50-61, such as SEQ ID NOs: 56, 57, 58, or 59.

[0535] 202. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 20, a CDR2 comprising the sequence of sequence A7, and a CDR1 comprising the sequence of SEQ ID NO: 56.

[0536] 203. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 21, a CDR2 comprising the sequence of sequence A8, and a CDR1 comprising the sequence of SEQ ID NO: 57.

[0537] 204. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 22, a CDR2 comprising the sequence of sequence A9, and a CDR1 comprising the sequence of SEQ ID NO: 58.

[0538] 205. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 23, a CDR2 comprising the sequence of sequence A10, and a CDR1 comprising the sequence of SEQ ID NO: 59.

[0539] 206. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 6, and a VL region comprising the CDR1, CDR2 and CDR3 sequences defined in clause 202.

[0540] 207. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises CDR1, CDR2 and CDR3 sequences as defined in clause 7 and a VL region comprising CDR1, CDR2 and CDR3 sequences as defined in clause 203.

[0541] 208. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises CDR1, CDR2 and CDR3 sequences as defined in clause 8 and a VL region comprising CDR1, CDR2 and CDR3 sequences as defined in clause 204.

[0542] 209. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises CDR1, CDR2 and CDR3 sequences as defined in clause 9 and a VL region comprising CDR1, CDR2 and CDR3 sequences as defined in clause 205.

[0543] 210. The anti-Vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62-85.

[0544] 211. The anti-vδ1 antibody or fragment thereof of clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62-73.

[0545] 212. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 62, 63, 64, 68, 69, 70 or 71.

[0546] 213. The anti-vδ1 antibody or fragment thereof of clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74-85.

[0547] 214. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising an amino acid sequence having at least 80% sequence identity to any one of SEQ ID NOs: 74, 75, 76, 80, 81, 82 or 83.

[0548] 215. The anti-vδ1 antibody or fragment thereof according to clause 190, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising the amino acid sequence of SEQ ID NO: 62 and ...

Claims

1. An anti-vδ1 antibody or fragment thereof for use in a method for treating cancer, an infectious disease or an inflammatory disease.

2. the anti-Vδ1 antibody or fragment thereof CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; a CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26-37 and A1-A12 (of Table 3); and / or 2. The anti-vδ1 antibody or fragment thereof of claim 1, comprising one or more CDR1s comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 38 to 61.

3. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 2, a CDR2 comprising the sequence of SEQ ID NO: 26, and a CDR1 comprising the sequence of SEQ ID NO:

38.

4. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 4, a CDR2 comprising the sequence of SEQ ID NO: 28, and a CDR1 comprising the sequence of SEQ ID NO:

40.

5. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region comprising a CDR3 comprising the sequence of SEQ ID NO: 3, a CDR2 comprising the sequence of SEQ ID NO: 27, and a CDR1 comprising the sequence of SEQ ID NO:

39.

6. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 14, a CDR2 comprising the sequence of sequence A1, and a CDR1 comprising the sequence of SEQ ID NO:

50.

7. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 16, a CDR2 comprising the sequence of sequence A3, and a CDR1 comprising the sequence of SEQ ID NO:

52.

8. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VL region comprising a CDR3 comprising the sequence of SEQ ID NO: 15, a CDR2 comprising the sequence of sequence A2, and a CDR1 comprising the sequence of SEQ ID NO:

51.

9. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof comprises a VH region and a VL region, and the VH region and VL region are linked by a linker, such as a polypeptide linker.

10. The anti-Vδ1 antibody or fragment thereof according to claim 1, wherein the anti-Vδ1 antibody or fragment thereof binds to or competes with the same or essentially the same epitope as the antibody or fragment thereof defined in any one of claims 1 to 9.

11. The anti-vδ1 antibody or fragment thereof of claim 1, wherein the anti-Vδ1 antibody or fragment thereof binds to an epitope of the variable delta 1 (Vδ1) chain of the gamma delta T cell receptor (TCR) comprising one or more amino acid residues within amino acids 1 to 90 of SEQ ID NO:

1.

12. The anti-vδ1 antibody or fragment thereof of claim 11, wherein the epitope comprises at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO:

1.

13. The anti-vδ1 antibody or fragment thereof of claim 11 or 12, wherein the epitope comprises one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:

1.

14. The anti-vδ1 antibody or fragment thereof according to any one of claims 11 to 13, wherein the epitope consists of one or more amino acid residues within the amino acid regions 5 to 20 and 62 to 77; 50 to 64; 37 to 53 and 59 to 72; 59 to 77; or 3 to 17 and 62 to 69 of SEQ ID NO:

1.

15. The anti-vδ1 antibody or fragment thereof according to any one of claims 11 to 14, wherein the epitope comprises one or more amino acid residues within amino acid regions 5 to 20 and 62 to 77 of SEQ ID NO:

1.

16. The anti-vδ1 antibody or fragment thereof according to any one of claims 11 to 15, wherein the epitope comprises one or more amino acid residues within the amino acid region 50 to 64 of SEQ ID NO:

1.

17. The anti-vδ1 antibody or fragment thereof according to any one of claims 11 to 16, wherein the epitope comprises one or more amino acid residues within the amino acid regions 37-53 and 59-77 of SEQ ID NO:

1.

18. The anti-vδ1 antibody or fragment thereof according to any one of claims 11 to 17, wherein the epitope is an activation epitope of γδ T cells.

19. The anti-vδ1 antibody or fragment thereof according to claim 1, wherein the antibody or fragment thereof defined in any one of claims 1 to 18 is an scFv, Fab, Fab', F(ab')2, Fv, a variable domain (e.g., VH or VL), a diabody, a minibody, or a full-length antibody.

20. 20. The anti-vδ1 antibody or fragment thereof of claim 19, wherein the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

111.

21. 20. The anti-vδ1 antibody or fragment thereof of claim 19, wherein the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

112.

22. 20. The anti-vδ1 antibody or fragment thereof of claim 19, wherein the antibody or fragment thereof comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

116.

23. 1. An isolated multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first target antigen of said at least two target antigens is Vδ1; CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2 to 25; a CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 26-37 and A1-A12 (of Table 3); and / or An isolated multispecific antibody or fragment thereof, comprising one or more of CDR1 comprising a sequence having at least 80% sequence identity to any one of SEQ ID NOs: 38 to 61.

24. 24. The isolated multispecific antibody or fragment thereof of claim 23, comprising a VH region and a VL region, wherein the VH region and VL region are linked by a linker, such as a polypeptide linker.

25. 25. An isolated multispecific antibody or fragment thereof that binds to or competes with the same or essentially the same V51 epitope as an antibody or fragment thereof defined in any one of claims 23 or 24.

26. 1. An isolated human anti-TCR delta variable 1 multispecific antibody or fragment thereof that binds to at least two target antigens, wherein a first target antigen of said at least two target antigens is Vδ1, and wherein said multispecific antibody or fragment thereof that binds to an epitope of Vδ1 comprises one or more amino acid residues within amino acids 1 to 90 of SEQ ID NO:

1.

27. 133. The human isolated multispecific antibody or fragment thereof of claim 132, wherein the epitope comprises at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72, or 77 of SEQ ID NO:

1.

28. 28. The human isolated multispecific antibody or fragment thereof according to claim 26 or 27, wherein the epitope comprises one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:

1.

29. 29. The human isolated multispecific antibody or fragment thereof of claim 28, wherein the epitope consists of one or more amino acid residues within the amino acid regions 5-20 and 62-77; 50-64; 37-53 and 59-72; 59-77; or 3-17 and 62-69 of SEQ ID NO:

1.

30. 30. The human isolated multispecific antibody or fragment thereof according to any one of claims 26 to 29, wherein said epitope comprises one or more amino acid residues within amino acid regions 5 to 20 and 62 to 77 of SEQ ID NO:

1.

31. 31. The human isolated multispecific antibody or fragment thereof according to any one of claims 26 to 30, wherein said epitope comprises one or more amino acid residues within the amino acid region 50 to 64 of SEQ ID NO:

1.

32. 32. The human isolated multispecific antibody or fragment thereof according to any one of claims 26 to 31, wherein said epitope comprises one or more amino acid residues within the amino acid regions 37 to 53 and 59 to 77 of SEQ ID NO:

1.

33. 33. The human isolated multispecific antibody or fragment thereof according to any one of claims 26 to 32, wherein said epitope is an activating epitope of γδ T cells.

34. 27. The isolated multispecific antibody of claim 26, comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

111.

35. 27. The isolated multispecific antibody of claim 26, comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

112.

36. 27. The isolated multispecific antibody of claim 26, comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

116.

37. 37. The isolated multispecific antibody or fragment thereof according to any one of claims 23 to 36, wherein a second target antigen of the at least two target antigens is a target antigen selected from the group consisting of EGFR and CD19.

38. 38. The isolated multispecific antibody or fragment thereof according to any one of claims 23 to 37 for use as a medicament.

39. 38. The isolated multispecific antibody or fragment thereof according to any one of claims 23 to 37 for use in the treatment of cancer, an infectious disease or an inflammatory disease.

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