Anti-TCRδ chain variable region 1 antibody

Abstract

The present invention provides high affinity anti-TCR delta variable region 1 (anti-V delta 1) antibodies and antibody fragments thereof. The present invention also provides compositions and pharmaceutical compositions comprising such antibodies, as well as methods for producing such antibodies. The present invention also provides methods of therapeutic and medical use comprising the antibodies.

Description

[Technical Field]

[0001] The present invention relates to antibodies that specifically bind to T cell receptors on γδT cells, as well as fragments and variants thereof. [Background technology]

[0002] The growing interest in T-cell immunotherapy for cancer focuses on the apparent ability of subsets of CD8+ and CD4+ αβT cells to recognize cancer cells and mediate host-protective functionality when desuppressed by clinically mediated antagonisms of inhibitory pathways exerted particularly by PD-1, CTLA-4, and other receptors. However, αβT cells are MHC-restricted and can cause graft-versus-host disease.

[0003] Gamma delta T cells (γδ T cells) are a subset of T cells that express different characteristic γδ T cell receptors (TCRs) on their surface. These TCRs consist of one gamma (γ) chain and one delta (δ) chain, each undergoing chain rearrangement, but with 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 the non-functional genes TRGV10, TRGV11, TRGVA, and TRGVB. The most frequent TRDV gene segments encode Vδ1, Vδ2, and Vδ3, and also encode several V segments with both Vδ and Vα names (Adams et al., 296:30-40 (2015) Cell Immunol.). Human γδT cells can be broadly classified based on their TCR chains, as specific γ and δ types are widely, though not exclusively, found in one or more tissue types. For example, most blood-resident γδT cells express the Vδ2 TCR, commonly Vγ9Vδ2, while this is less common in tissue-resident γδT cells, such as those in the skin. Tissue-resident γδT cells more frequently use the Vδ1 TCR paired with the γ chain, for example, the Vδ1 TCR paired with Vγ4 in the intestines.

[0004] γδT cells play a crucial role in immune surveillance, recognizing malignant or transformed cells (such as cancer cells) based on stress marker patterns, and subsequently exerting potent and selective cytotoxicity. Therefore, γδT cells can act as orchestrators of the immune response. In vivo modulation of these cells may lead to increased immunogenicity even in low-mutagram-burden tumors, which have proven difficult to treat with other immunotherapies. γδT cell recognition of tumors is not dependent on any single tumor antigen; therefore, γδT cell modulators have potential in a wide range of disease indications, including both hematological and solid malignancies. The γδT cell recognition mechanism is not MHC-restricted.

[0005] The authors of WO2019147735 hypothesize that some γδ cells have tumorigenic activity or inhibit the anti-cancer immune response mediated by αβ T cells. They hypothesize that γδ T cells are immunosuppressants and therefore propose that antibodies should be used to eliminate, inhibit, or block them in cancer.

[0006] However, despite the prevailing view that anti-γδ antibodies negatively regulate the function of γδ cells by blocking or killing them, a positive correlation has been found between γδT cell infiltration and patient prognosis and / or survival.

[0007] Compared to αβ TCR receptor / ligand interactions, our understanding of vδ1 TCR receptor / ligand interactions is limited. Due to this lack of understanding, antibodies that recognize vδ1 TCRs have primarily served as exploratory tools for investigating this interaction. Typically, such tools are crude blocking antibodies suggesting that TCR receptor / ligand interactions result in the blockade, suppression, or removal of vδ1+ cells. For example, the tool antibodies TS8.2 and TS-1 have been used as anti-γδ blocking antibodies in studies, suggesting that these antibodies reduce the cytotoxicity of vδ1 cells. These studies, combined with others, suggest that such anti-vδ1 antibodies would not be useful in favorably regulating vδ1 cell cytotoxicity in in-vivo disease settings; therefore, there is a need for antibodies that increase, rather than decrease, vδ1 cytotoxicity.

[0008] To utilize γδT cells in immunotherapy, either a means of growing the cells in situ in vivo or a means of harvesting them and growing them ex vivo before reinjection is required. The latter approach, using the addition of exogenous cytokines, has been previously described. See, for example, WO2017 / 072367 and WO2018 / 212808. Methods for growing a patient's own γδT cells using pharmacologically modified forms of hydroxymethylbuta-2-enylpyrophosphate (HMBPP) or clinically approved aminobisphosphonates have been described. With these approaches, more than 250 cancer patients have been treated seemingly safely, although the occurrence of complete remission is extremely rare. Nevertheless, there is still a need for activators that have demonstrated the ability to grow a large number of γδT cells.

[0009] Furthermore, conjugates or activators that can preferentially target, bind to, recognize, specifically modulate, or increase the number of Vδ1+ cells in vivo are highly desirable as pharmaceutical agents.

[0010] However, while there are drugs that can potentially modulate Vδ2+ cells, including aminobisphosphonates such as Zometa® (zoledronic acid), these drugs are primarily designed to delay bone resorption. Furthermore, regardless of the above-mentioned modulation of Vδ2+, there is a need to develop drugs specifically designed to bind to, target, modulate, activate, or increase the number of Vδ1+ cells. This is because, for example, repeated modulation of Vδ2+ can lead to a persistent and progressive exhaustion phenotype.

[0011] Furthermore, considering that Vδ1+ cells are primarily tissue-resident, an ideal agent capable of modulating Vδ1+ would also exhibit smaller, undesirable "off-target" effects and rapid renal clearance. Typically, these undesirable effects can manifest when small-molecule chemicals are used. For example, the aforementioned aminobisphosphonates, which have been shown to modulate a distinct class of Vδ2+ cells (as a secondary effect to their primary modulatory effect on bone), are associated with nephrotoxicity, which manifests as worsening of renal function and potential renal failure (e.g., Markowitz et al. (2003) Kidney Int. 64(1):281-289). Additional undesirable effects of Zometa listed by the European Medicines Agency include anemia, hypersensitivity reactions, hypertension, atrial fibrillation, myalgia, generalized pain, malaise, increased blood urea, vomiting, joint swelling, and chest pain.

[0012] The in vivo environment in which vδ1+ cells are found must also be considered. For example, it has been previously shown that non-hematopoietic tissue-resident γδ T cells show a strong proliferative response when initially isolated from tissue, but only when they are not in direct cell contact with autologous fibroblasts. It has been found that non-hematopoietic tissue-resident T cells (γδ T cells) must be isolated from non-hematopoietic cells (e.g., stromal cells, especially fibroblasts) in order to function. This is because direct contact between lymphocytes and stromal or epithelial cells appears to inhibit the proliferation of tissue-resident γδ T cells. The observation that pre-activation cells exist in a further suppressed state in vivo is another reason why vδ1 cells have not been considered a promising therapeutic target until now. In fact, until the findings described herein, no method has been devised to suitably and selectively regulate blood and tissue vδ1+ cells in vivo when these cells are typically considered to be in a “resting,” “pre-activation,” or “inactivated” state.

[0013] Various formats of bispecific and multispecific antibodies have been developed for a wide range of therapeutic applications. Bispecific and multispecific antibodies can be classified into overlapping but distinct classes based on the type of biological target and mode of action. For example, such multispecific antibodies can be classified into classes such as cytotoxic effector cell redirectors (also known as bispecific T cell recruiting antibodies, bispecific T cell engagers, TCEs, or BiTEs) and dual immunomodulators (DIs).

[0014] T-cell engagers (TCEs) are intended to enhance a patient's immune response to tumors by targeting T cells to tumor cells or vice versa. They function by targeting a primary epitope (usually CD3) and a secondary epitope, which is a cancer-associated antigen such as a cancer antigen or tumor-associated antigen (TAA), on the T cell receptor complex of T cells. Such antibodies co-localize on tumor cells and T cells to promote tumor cell killing. Examples of BiTEs include blinatumomab, a CD3 x CD19 bispecific antibody; catumaxomab, a CD3 x EpCAM bispecific antibody; and erzmaxomab, a CD3 x HER2 bispecific antibody. TCEs such as BiTEs are usually supplied in scFv format, but other formats are also available. For example, BiKEs are similar to BiTEs but target CD16 on NK cells instead of CD3.

[0015] The T cell receptor is described as the most complex receptor structure in the mammalian immune system. It constitutes a transmembrane multiprotein receptor complex containing several CD3 chains in close proximity to the T cell receptor. For example, in mammals, a typical such complex includes the T cell receptor, CD3γ chain, CD3δ chain, and two CD3ε chains. These chains associate with the T cell receptor (TCR) near the bound ζ (zeta) chain, subsequently generating a typical activation signal in T lymphocytes. However, alternative complexes have also been reported. For example, T cell receptor complexes containing the T cell receptor and ζ chain homodimers have been described. Additional co-receptors, such as CD4 and CD8, may also assist TCR function.

[0016] Regardless of the receptor complex composition, it has been established that the above complex converts cell surface binding events into intracellular phosphorylation signaling cascades. These phosphorylation events ultimately lead to the activation of transcription factors such as NFAT and NFκB, which in turn increase the expression of cytokines and effector proteins such as granzymes and perforins.

[0017] However, while the use of such TCEs to treat cancer remains a compelling concept, to date, and even after 30 years of coordinated efforts to advance TCEs in early clinical development, many of these bispecific antibodies have shown subpar safety, efficacy, and manufacturability profiles. In fact, as of January 2020, blinatumomab remains the only TCE that has not been withdrawn after approval. This TCE multispecific antibody fragment binds to the T cell receptor complex with its first binding arm and to the CD19 target with its second binding arm.

[0018] Bispecific T-cell recruiting antibodies are described in Lejeune et al., 2020, Front Immunol., 11:762. However, existing bispecific antibodies in this category, particularly those that recruit T cells via CD3 binding, have significant off-target effects that result in severe adverse effects, given the potency of the CD3 antigen as a signaling molecule and its ubiquity in the patient's T-cell population. Therefore, systemic delivery (e.g., intravenous) is practically impossible for examples of CD3-targeting bispecific antibodies such as catumaxomab (now retracted). Instead, more restricted deliveries, such as intraoperative, intraperitoneal, and intraperitoneal delivery, are more frequently intended. This limits the selectivity and use of the above bispecific drugs as pharmaceuticals. In fact, even anti-CD3 antibodies with attenuated effector function (i.e., T-cell complex engagers that target CD3 but are not bispecific) are difficult to deliver intravenously due to associated toxicity. For example, foralumab, an anti-CD3 antibody, is now almost always intended for oral delivery (e.g., in the treatment of gastrointestinal diseases) to limit exposure and reduce toxicity.

[0019] Many of the current problems observed with such TCEs in early clinical trials are often attributed to the high-affinity T cell complex binding domains used. Furthermore, it has been suggested that this is because the designers of these TCEs did not adequately consider the low affinity of the TCR complex binding event in nature, and severe dose-limiting toxicity hindered their development, resulting in an extremely narrow therapeutic range. In this regard, it has been highlighted that many early TCE drug developers relied on three anti-CD3 T cell complex binding domains derived from OKT3, SP34, and UCHT1. All of these original binding domains bind with relatively high affinities in the nM range of single to low double digits, equivalent to an affinity approximately 1,000 times higher than the binding event in nature. As a result, it has been suggested that T cell activation is affected in a significantly different (and often unfavorable) way compared to the binding of the T cell receptor complex in nature. For example, TCE developers using platforms based on the higher-affinity OKT3 may be hampered by the fact that OKT3 induces T cell apoptosis in the presence of IL-2.

[0020] For these reasons, the binding of lower affinity T cell complexes has become an important consideration when determining the design parameters for bispecific antibody therapeutics that associate with T cells.

[0021] Another issue in designing the above TCEs is the need to attenuate Fc function. In fact, since the binding of Fc to the Fcγ receptor (FcγR) triggers the activation of immune effector cells, TCEs typically require complete suppression of Fc-mediated effector function to maximize therapeutic efficacy and minimize off-target toxicity. Indeed, the vast majority of CD3-targeting bispecific antibodies currently used in clinical practice either have an Fc domain with reduced binding activity to FcγR or are bispecific fragments that intentionally omit the Fc region. Generally, TCEs with unattenuated Fc function are expected to induce antibody-dependent cell-mediated cytotoxicity (ADCC), thereby depleting the antibody-recognized γδ T cell population. However, again, attenuating such functions to avoid toxicity / safety complexities, for example by involving CD16+, CD32+, or CD64+ immune cells, or by reducing the bispecific half-life (e.g., by using smaller bispecific antibody fragments such as BITE), may also attenuate potentially important efficacy aspects. Methods that reduce the interaction between FcγR and TCE (e.g., by using an IgG format designed to reduce such interaction) are expected to reduce the immobilization of TCE mediated by Fc and reduce TCR clustering by crosslinking with immobilized TCE.

[0022] To address some, though not all, of these issues, numerous companies, including Xencor (Pasadena, CA), Macrogenics (Gaithersburg, MD), and Genentech (San Francisco, CA), have very recently reported reducing the binding affinity of the T cell receptor complex binding arm in their respective TCE platforms. However, this reduction in binding affinity can lead to decreased efficacy and selectivity in terms of TCE design and functionality. For example, it has now been demonstrated that the affinity of the binding domain in such TCEs is a determinant of the in vivo distribution profile. Specifically, the distribution of TCEs is usually observed to be biased towards the target with the highest affinity. Therefore, reducing the affinity of the TCE binding domain to the T cell complex generally leads to a biased distribution away from T cells, which are precisely the cells needed to produce efficacy for such TCEs. This is partly why the therapeutic range of TCEs is said to be "extremely narrow."

[0023] Furthermore, significant hurdles remain for immunomodulatory therapies, particularly in solid tumors. For example, the current state-of-the-art approach involves CD3-targeted bispecific drugs, where the first domain binds to CD3 and the second domain targets TAAs. However, these have often proven problematic. For instance, Middelburg J, et al. Overcoming Challenges for CD3-Bispecific Antibody Therapy in Solid Tumors. Cancers. 2021;13(2):287 summarizes some of these hurdles for multispecific immunomodulatory moieties that associate with T cells in solid tumor space, including: i. On-target / off-tumor toxicity issues: For example, the authors emphasize that TAAs in solid tumors are often expressed in the tissues of healthy organs, which can lead to organ failure that may result in immunopathology and death, as demonstrated in preclinical studies in mice using CD3 bispecific drugs. ii. The availability of effector cells in the tumor microenvironment (TME): For example, normal CD3+ "naive" T cells are mainly found in the blood and lymphatic systems without being further activated by dendritic cells. iii. Quality of tumor-infiltrating lymphocytes (TILs), considering that TILs may include CD3+ Treg cells and / or exhausted CD3+ cells: For example, CD3-targeted bispecific drugs may induce TIL apoptosis through activation-induced cell death, which hinders a potent antitumor response.

[0024] Therefore, there is a need for improved multispecific immunomodulatory drugs in which at least one binding domain binds to T cells and at least one second binding domain targets TAAs.

[0025] There is also a need for improved drugs specifically designed to target Vδ1+ cells and treat infections, autoimmune conditions, and cancer. Specifically, there is a need for drugs that can be administered to improve the signs and symptoms of disease by specifically binding to, targeting, specifically activating, specifically enhancing the proliferation and / or cytotoxic activity of Vδ1+ cells, or specifically inhibiting the activation of Vδ1+ cells. [Overview of the project]

[0026] This invention provides a high-affinity anti-TCRδ variable region 1 (anti-Vδ1) antibody and its antibody fragment. The antibody of this invention has a favorable functional profile. In particular, unlike conventional anti-Vδ1 antibodies that focus on Vδ1 T cell depletion, the antibody of this invention is useful for Vδ1 T cell activation. The antibody of this invention can induce downregulation of the TCR on the T cells to which they bind, but does not cause Vδ1 T cell depletion; rather, it stimulates T cells, and therefore may be useful in therapeutic situations where activation of this compartment of T cells is beneficial. Vδ1 T cell activation is evident from downregulation of the TCR, activation markers such as CD25, Ki67, the degranulation marker CD107a, NCR (innate cytotoxic receptor), and / or changes in 4-1BB. Vδ1 T cell activation then triggers the release of inflammatory cytokines such as INFγ and TNFα, promoting immune licensing. Surprisingly, antibodies with appropriately high affinity for TRDV1 induce increased Vδ1 T cell killing, and unlike antibodies targeting (e.g.) CD3, high-affinity antibodies can be supplied without the side effects associated with large-scale activation by CD3. As a result, high-affinity antibodies can induce a potent immunostimulatory effect by tumor-infiltrating lymphocytes (TILs). This can be achieved with minimal exhaustion or killing of Vδ1 cells. Therefore, the antibodies of the present invention can be considered agonist antibodies.

[0027] According to a first aspect of the present invention, VHCDR3 containing an amino acid sequence having at least 80% sequence identity to any of SEQ ID NOs. 55-78; and / or VLCDR3 contains an amino acid sequence having at least 80% sequence identity to any of sequence numbers 82-105, or VHCDR3 containing an amino acid sequence having at least 80% sequence identity to any of SEQ ID NOs: 133-143; and / or An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, which contains VLCDR3 having an amino acid sequence having at least 80% sequence identity to any of SEQ ID NOs. 147 to 157.

[0028] According to a second aspect of the present invention, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, HCDR1 sequence containing the sequence of GDSVSSKSX1A (sequence number 158); HCDR2 sequence containing sequence number 53; An HCDR3 sequence containing the sequence X2WX3X4X5X6DX7 (sequence number 162), but not sequence number 54; LCDR1 sequence containing sequence number 79; LCDR2 sequence containing sequence number 80; and QQX8YX9X 10 X 11 X 12 X 13 An LCDR3 sequence containing the sequence T (sequence number 166), wherein the LCDR3 sequence is not sequence number 81, and the LCDR3 sequence contains the said LCDR3 sequence, X1~X 13 Each of these is a naturally occurring amino acid, the anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, or An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, HCDR1 sequence containing sequence number 130; HCDR2 sequence containing sequence number 131; An HCDR3 sequence containing the sequence X1X2YX3X4AFDI (sequence number 183), but not sequence number 132; LCDR1 sequence containing sequence number 144; LCDR2 sequence containing sequence number 145; and An LCDR3 sequence containing the sequence QQX5X6X7X8LX9T (sequence number 186), wherein the LCDR3 sequence is not sequence number 146, and includes the said LCDR3 sequence, The anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, wherein each of X1 to X9 is a naturally occurring amino acid.

[0029] In a third aspect of the present invention, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof is provided, wherein the anti-Vδ1 antibody or antigen-binding fragment is an affinity-matured variant of a parent anti-Vδ1 antibody or its antigen-binding fragment, wherein the parent anti-Vδ1 antibody or its antigen-binding fragment comprises a VH sequence containing the amino acid sequence of SEQ ID NO: 1 and a VL sequence containing the amino acid sequence of SEQ ID NO: 26, or the parent anti-Vδ1 antibody or its antigen-binding fragment comprises a VH sequence containing the amino acid sequence of SEQ ID NO: 106 and a VL sequence containing the amino acid sequence of SEQ ID NO: 118. Optionally, the first residue of the VH sequence in any affinity-matured variant may be Q or E. Optionally, the first two residues of the VL sequence in any affinity-matured variant may be absent in any affinity-matured variant compared to the original sequence.

[0030] In a fourth aspect of the present invention, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, wherein the anti-Vδ1 antibody or its antigen-binding fragment is an affinity-mature variant of the parental anti-Vδ1 antibody or its antigen-binding fragment, and the parental anti-Vδ1 antibody or its antigen-binding fragment is a) VH containing the amino acid sequence of SEQ ID NO: 273 and VL containing the amino acid sequence of SEQ ID NO: 282; b) VH containing the amino acid sequence of SEQ ID NO: 274 and VL containing the amino acid sequence of SEQ ID NO: 283; c) VH containing the amino acid sequence of SEQ ID NO: 275 and VL containing the amino acid sequence of SEQ ID NO: 284; d) VH containing the amino acid sequence of SEQ ID NO: 276 and VL containing the amino acid sequence of SEQ ID NO: 285; e) VH containing the amino acid sequence of SEQ ID NO: 277 and VL containing the amino acid sequence of SEQ ID NO: 286; f) VH containing the amino acid sequence of SEQ ID NO: 278 and VL containing the amino acid sequence of SEQ ID NO: 287; g) VH containing the amino acid sequence of SEQ ID NO: 279 and VL containing the amino acid sequence of SEQ ID NO: 288; h) VH containing the amino acid sequence of SEQ ID NO: 280 and VL containing the amino acid sequence of SEQ ID NO: 289; i) VH containing the amino acid sequence of SEQ ID NO: 281 and VL containing the amino acid sequence of SEQ ID NO: 290; or j) Includes VH containing the amino acid sequence of SEQ ID NO: 312 and VL containing the amino acid sequence of SEQ ID NO: 313.

[0031] A fifth aspect of the present invention provides an anti-TCRδ variable region 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof, comprising a κ light chain variable sequence wherein the residue at position 74 of the κ light chain variable sequence is not serine according to the IMGT numbering system. In some embodiments, serine may be substituted with an amino acid at position 74 in a non-human germline. In some embodiments, the substitution may be a non-conservative mutation, such as substituting serine with a nonpolar amino acid. In some embodiments, serine may be substituted with leucine.

[0032] In a sixth aspect of the present invention, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, which specifically binds to the variable δ1 (Vδ1) chain of the γδT cell receptor (TCR) and competes with any of the antibodies or antigen-binding fragments of the first to fifth aspects of the present invention for binding to the variable δ1 (Vδ1) chain of the γδT cell receptor (TCR).

[0033] A seventh aspect of the present invention provides a polynucleotide sequence encoding an anti-Vδ1 antibody or its antigen-binding fragment. For example, a polynucleotide sequence is provided that includes a sequence having at least 70% sequence identity with SEQ ID NOs. 199-222, 224-247, 249-259, or 261-271, encoding an anti-Vδ1 antibody or its antibody-binding fragment.

[0034] An eighth aspect of the present invention provides an expression vector comprising the polynucleotide sequence of the present invention. A host cell comprising the polynucleotide sequence of the present invention or the expression vector of the present invention is also provided. A method for producing any antibody or antigen-binding fragment thereof of the present invention is also provided, comprising culturing the host cell of the present invention in a cell culture medium.

[0035] In further aspects of the present invention, a composition comprising the antibody of the present invention or an antibody-conjugated fragment thereof is provided. A pharmaceutical composition comprising the antibody of the present invention or an antibody-conjugated fragment thereof and a pharmaceutically acceptable diluent or carrier is also provided. The composition and the pharmaceutical composition may optionally further comprise one or more additional therapeutic agents.

[0036] In a further aspect of the present invention, a kit is provided comprising the anti-Vδ1 antibody or antibody-conjugated fragment of the present invention, or the pharmaceutical composition of the present invention, and optionally comprising instructions for use and / or additional therapeutic agents.

[0037] Further aspects of the present invention provide a method for treating a disease or disorder of interest, comprising administering an anti-Vδ1 antibody or antibody-conjugated fragment of the present invention, or a pharmaceutical composition of the present invention, to the subject. Also provided is a method for modulating an immune response in a subject, comprising administering an anti-Vδ1 antibody or antibody-conjugated fragment of the present invention, or a pharmaceutical composition of the present invention, to the subject. The administration of the antibody to the subject may be in a therapeutically effective dose.

[0038] In yet another aspect of the present invention, a method for mutating an antibody or an antigen-binding fragment thereof is provided, comprising providing an antibody comprising a κ light chain having serine at position 74 according to the IMGT numbering system, and substituting (e.g., mutating) the serine with a different amino acid. In some embodiments, the serine may be substituted with the 74th amino acid of a non-human germline. In some embodiments, the substitution may be a non-conservative mutation, such as substituting serine with a nonpolar amino acid. In some embodiments, the serine may be substituted with leucine.

[0039] A further aspect of the present invention relates to a method for preparing an anti-TCRδ variable region 1 (anti-Vδ1) antibody variant or its antigen-binding fragment, a) VH containing the amino acid sequence of SEQ ID NO: 1 and VL containing the amino acid sequence of SEQ ID NO: 26; b) VH containing the amino acid sequence of SEQ ID NO: 106 and VL containing the amino acid sequence of SEQ ID NO: 118; c) VH containing the amino acid sequence of SEQ ID NO: 273 and VL containing the amino acid sequence of SEQ ID NO: 282; d) VH containing the amino acid sequence of SEQ ID NO: 274 and VL containing the amino acid sequence of SEQ ID NO: 283; e) VH containing the amino acid sequence of SEQ ID NO: 275 and VL containing the amino acid sequence of SEQ ID NO: 284; f) VH containing the amino acid sequence of SEQ ID NO: 276 and VL containing the amino acid sequence of SEQ ID NO: 285; g) VH containing the amino acid sequence of SEQ ID NO: 277 and VL containing the amino acid sequence of SEQ ID NO: 286; h) VH containing the amino acid sequence of SEQ ID NO: 278 and VL containing the amino acid sequence of SEQ ID NO: 287; i) VH containing the amino acid sequence of SEQ ID NO: 279 and VL containing the amino acid sequence of SEQ ID NO: 288; j) VH containing the amino acid sequence of SEQ ID NO: 280 and VL containing the amino acid sequence of SEQ ID NO: 289; k) VH containing the amino acid sequence of SEQ ID NO: 281 and VL containing the amino acid sequence of SEQ ID NO: 290; or l) To provide a parent antibody comprising VH containing the amino acid sequence of SEQ ID NO: 312 and VL containing the amino acid sequence of SEQ ID NO: 313. The provided method includes subjecting an antibody to affinity maturation, wherein the produced antibody has a stronger affinity for the variable δ1 (Vδ1) chain of the γδ T cell receptor (TCR) compared to the parent antibody.

[0040] A further aspect of the present invention provides a method for preparing a pharmaceutical composition, comprising providing an antibody prepared according to a method for preparing an anti-TCRδ variable portion 1 (anti-Vδ1) antibody variant or antigen-binding fragment of the present invention, and co-formulating the antibody with at least one pharmaceutically acceptable diluent or carrier.

[0041] In yet another aspect of the present invention, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment for medical use, or a pharmaceutical composition of the present invention, or a kit of the present invention is provided. The use of the anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment in the manufacture of a drug is also provided. [Brief explanation of the drawing]

[0042] [Figure 1] ELISA detection of directly coated antigens using anti-Vδ1Ab (REA173, Miltenyi Biotec). Detection was observed only with antigens containing the Vδ1 domain. The leucine zipper (LZ) format appeared to be more potent than the Fc format, which is consistent with cell-based flow-competitive assays (data not shown). [Figure 2] Polyclonal phage DELFIA data for DV1 selection. (A) Heterodimer selection: Heterodimer LZ TCR format in rounds 1 and 2, deselection of heterodimer LZ TCR in both rounds. (B) Homodimer selection: Round 1 was performed using homodimer Fc fusion TCR to deselect human IgG1 Fc, followed by Round 2 for heterodimer LZ TCR to deselect heterodimer LZ TCR. Each graph contains two bars for each target to represent selection from different libraries. [Figure 3] IgG capture: Left) Sensorgram of the interaction between anti-L1 IgG and L1, Right) Steady-state fit when available. All experiments were performed at room temperature using a MASS-2 instrument. Steady-state fitting follows Langmuir 1:1 binding. [Figure 4]Results of TCR downregulation assays (using THP-1 pre-filled antibodies) 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 injury assays (THP-1 flow-based 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 7] Total cell count in Experiment 1 of Example 10. Samples were cultured with various concentrations of the anti-Vδ1 antibody described herein and compared with samples cultured with a comparator antibody or a control. The graphs show the total cell counts on (A) day 7, (B) day 14, and (C) day 18. [Figure 8] Analysis of Vδ1 T cells in Experiment 1 of Example 10. The graph shows (A) the percentage of Vδ1 T cells, (B) the number of Vδ1 T cells, and (C) the Vδ1 change ratio in the sample on day 18. [Figure 9A]Total cell count in Experiment 2 of Example 10. Samples were cultured with various concentrations of the anti-Vδ1 antibody described herein and compared with samples cultured with a comparator antibody or a control. The graphs show the total cell count on (A) day 7, (B) day 11, (C) day 14, and (D) day 17. [Figure 9B] Total cell count in Experiment 2 of Example 10. Samples were cultured with various concentrations of the anti-Vδ1 antibody described herein and compared with samples cultured with a comparator antibody or a control. The graphs show the total cell count on (A) day 7, (B) day 11, (C) day 14, and (D) day 17. [Figure 9C] Total cell count in Experiment 2 of Example 10. Samples were cultured with various concentrations of the anti-Vδ1 antibody described herein and compared with samples cultured with a comparator antibody or a control. The graphs show the total cell count on (A) day 7, (B) day 11, (C) day 14, and (D) day 17. [Figure 9D] Total cell count in Experiment 2 of Example 10. Samples were cultured with various concentrations of the anti-Vδ1 antibody described herein and compared with samples cultured with a comparator antibody or a control. The graphs show the total cell count on (A) day 7, (B) day 11, (C) day 14, and (D) day 17. [Figure 10] Analysis of Vδ1 T cells in Experiment 2 of Example 10. The graph shows (A) the percentage of Vδ1 T cells, (B) the number of Vδ1 T cells, and (C) the Vδ1 change ratio in the sample on day 17. [Figure 11] Cell composition analysis. The cell types present in the sample (including non-Vδ1 cells) were measured on day 17 of Experiment 2. Cells were collected, and the surface expression of Vδ1, Vδ2, and αβTCR was analyzed by flow cytometry. Percentage values ​​are also shown in Table 12. [Figure 12] SYTOX flow injury assay results. Cell functionality was tested using the SYTOX flow injury assay, and the results are presented for (A) Experiment 1 at day 14 using cells with a 10:1 effector-to-target (E:T) ratio, and (B) Experiment 2 at day 17 (after freeze-thaw) using cells with 1:1 and 10:1 E:T ratios. [Figure 13] Total cell count after freeze-thawing. The graph shows the total cell count after 7 days of cell culture following freeze-thawing for cultures that were contacted with B07, C08, E07, G04, or OKT-3 antibodies before freezing. [Figure 14] Monitoring of cell growth. The total cell count of cells cultured after freezing and thawing was monitored until day 42. [Figure 15] Binding equivalence study of modified anti-Vδ1 antibody [Figure 16] Equivalence studies of anti-Vδ1 antibody binding to human germline Vδ1 antigen and its polymorphic variants. [Figure 17] Anti-Vδ1 antibody induced elevated cytokine secretion levels in Vδ1+ cells. Tissue-derived γδT cells were incubated with the indicated antibody. (A) Observed TNF-alpha levels. (B) Observed IFN-gamma levels. [Figure 18] Anti-Vδ1 antibodies induced elevated granzyme B levels / activity in Vδ1+ cells. Cancer cells were co-cultured for 1 hour with tissue-derived γδT cells in a constant 1:20 T:E ratio and with the indicated antibodies. The results reveal the amount of granzyme B detected in cancer cells at the end of co-culture. [Figure 19] Anti-Vδ1 antibodies conferred regulation and proliferation of immune cells in human tissue. Human skin punch biopsies (from 5 different donors) were incubated with the indicated antibodies in culture for 21 days. (A) Number of viable pan-γδ+ cells. (B) Number of viable Vδ1+ cells. (C) Percentage of viable double-positive Vδ1+CD25+ cells. [Figure 20] Anti-Vδ1 antibodies conferred regulation and proliferation of tumor-infiltrating lymphocytes (TILs) in human tumors. Studies on renal cell carcinoma (RCC) + / - cells: (A) Multiplier of TIL Vδ1+ cells. (B) Total number of TIL Vδ1+ cells. (C) Examples of gating strategies. (D) Comparison of cell surface phenotypic profiles of TIL Vδ1+ cells. (E) Analysis of TIL Vδ1-negative gate fractions. [Figure 21A]Anti-Vδ1 antibodies conferred enhanced Vδ1+-mediated cytotoxicity and disease-cell-specific cytotoxicity. Cytotoxicity / efficacy assays in a model system including tripartite cultures of Vδ1+ effector cells, THP-1 monocytic cancer cells, and healthy, non-disease primary monocytes. (A) Quantification of the number of THP-1 and monocyte cells in triple co-culture using γδT cells in the presence of anti-Vδ1 mAb or control. (B) Bar graphs highlighting the window between disease-specific injury and disease-free health: Left bar graph; increase in injury ratio of disease cells (THP-1) compared to injury of non-disease cells (primary human monocytes); Right bar graph; same data, but expressed as injury enhancement rate compared to control. (C) Tabular results summarizing the improvement in Vδ1+ effector cell-toxicity efficacy of THP-1 target cells ± mAb. (D) Tabular results of EC50 values ​​calculated from Figure (A), expressed as the number of γδ T cells required to induce 50% THP-1 cell damage. [Figure 21B] Anti-Vδ1 antibodies conferred enhanced Vδ1+-mediated cytotoxicity and disease-cell-specific cytotoxicity. Cytotoxicity / efficacy assays in a model system including tripartite cultures of Vδ1+ effector cells, THP-1 monocytic cancer cells, and healthy, non-disease primary monocytes. (A) Quantification of the number of THP-1 and monocyte cells in triple co-culture using γδT cells in the presence of anti-Vδ1 mAb or control. (B) Bar graphs highlighting the window between disease-specific injury and disease-free health: Left bar graph; increase in injury ratio of disease cells (THP-1) compared to injury of non-disease cells (primary human monocytes); Right bar graph; same data, but expressed as injury enhancement rate compared to control. (C) Tabular results summarizing the improvement in Vδ1+ effector cell-toxicity efficacy of THP-1 target cells ± mAb. (D) Tabular results of EC50 values ​​calculated from Figure (A), expressed as the number of γδ T cells required to induce 50% THP-1 cell damage. [Figure 21C]Anti-Vδ1 antibodies conferred enhanced Vδ1+-mediated cytotoxicity and disease-cell-specific cytotoxicity. Cytotoxicity / efficacy assays in a model system including tripartite cultures of Vδ1+ effector cells, THP-1 monocytic cancer cells, and healthy, non-disease primary monocytes. (A) Quantification of the number of THP-1 and monocyte cells in triple co-culture using γδT cells in the presence of anti-Vδ1 mAb or control. (B) Bar graphs highlighting the window between disease-specific injury and disease-free health: Left bar graph; increase in injury ratio of disease cells (THP-1) compared to injury of non-disease cells (primary human monocytes); Right bar graph; same data, but expressed as injury enhancement rate compared to control. (C) Tabular results summarizing the improvement in Vδ1+ effector cell-toxicity efficacy of THP-1 target cells ± mAb. (D) Tabular results of EC50 values ​​calculated from Figure (A), expressed as the number of γδ T cells required to induce 50% THP-1 cell damage. [Figure 21D] Anti-Vδ1 antibodies conferred enhanced Vδ1+-mediated cytotoxicity and disease-cell-specific cytotoxicity. Cytotoxicity / efficacy assays in a model system including tripartite cultures of Vδ1+ effector cells, THP-1 monocytic cancer cells, and healthy, non-disease primary monocytes. (A) Quantification of the number of THP-1 and monocyte cells in triple co-culture using γδT cells in the presence of anti-Vδ1 mAb or control. (B) Bar graphs highlighting the window between disease-specific injury and disease-free health: Left bar graph; increase in injury ratio of disease cells (THP-1) compared to injury of non-disease cells (primary human monocytes); Right bar graph; same data, but expressed as injury enhancement rate compared to control. (C) Tabular results summarizing the improvement in Vδ1+ effector cell-toxicity efficacy of THP-1 target cells ± mAb. (D) Tabular results of EC50 values ​​calculated from Figure (A), expressed as the number of γδ T cells required to induce 50% THP-1 cell damage. [Figure 22A]The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22B] The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22C]The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22D] The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22E]The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22F] The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22G]The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22H] The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22I]The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 22J] The polyspecific antibody conferred enhanced Vδ1+ effector cell-mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (A-D) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without (+ / -) polyspecific antibodies containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, which combines the anti-Vδ1 VL+VH binding domain (for the first target) with the CH1-CH2-CH3 domain of the anti-EGFR binding site (for the second target). (E~H) Examples of co-culture of Vδ1+ effector cells and A-431 cancer cells with or without a multispecific antibody containing an anti-Vδ1 × anti-TAA(EGFR) bispecific binding site, in which the anti-Vδ1 binding domain (for the first target) contains a full-length antibody (VH-CH1-CH2-CH3 / VL-CL), which is then combined with an anti-EGFR cetuximab-derived scFv binding site (for the second target). (I~J) Alternative approaches to express the data: Percentage of improvement in Vδ1+ effector cytotoxicity against EGFR+ cells relative to the constituent parts, as conferred by the multispecific antibody. [Figure 23A]The polyspecific antibody conferred enhanced Vδ1+-mediated cytotoxicity and disease cell-specific cytotoxicity. Targeting of hematopoietic disease-related antigens. (A) E:T ratio required to induce 50% Raji cell injury. (B) Percentage improvement by addition of Vδ1-CD19 polyspecific antibody. [Figure 23B] The polyspecific antibody conferred enhanced Vδ1+-mediated cytotoxicity and disease cell-specific cytotoxicity. Targeting of hematopoietic disease-related antigens. (A) E:T ratio required to induce 50% Raji cell injury. (B) Percentage improvement by addition of Vδ1-CD19 polyspecific antibody. [Figure 24A] Phage selection rounds. (A) Phage selection rounds 1-3 for the ADT1-7 library. (B) Phage selection rounds 1-3 for the ADT1-4 library. (C) Phage selection rounds 1-3 for the ADT1-4 library by selection strategies to isolate cynomolgus monkey cross-reactive binders. [Figure 24B] Phage selection rounds. (A) Phage selection rounds 1-3 for the ADT1-7 library. (B) Phage selection rounds 1-3 for the ADT1-4 library. (C) Phage selection rounds 1-3 for the ADT1-4 library by selection strategies to isolate cynomolgus monkey cross-reactive binders. [Figure 24C] Phage selection rounds. (A) Phage selection rounds 1-3 for the ADT1-7 library. (B) Phage selection rounds 1-3 for the ADT1-4 library. (C) Phage selection rounds 1-3 for the ADT1-4 library by selection strategies to isolate cynomolgus monkey cross-reactive binders. [Figure 25A] These are schematic diagrams of the selection of mature antibodies using mammalian display. (A) Schematic diagram of flow sorting of the ADT1-7 library. (B) Schematic diagram of flow sorting of ADT1-4 library 1 (human). (C) Schematic diagram of flow sorting of ADT1-4 library 2 (cynomolgus monkey). [Figure 25B]These are schematic diagrams of the selection of mature antibodies using mammalian display. (A) Schematic diagram of flow sorting of the ADT1-7 library. (B) Schematic diagram of flow sorting of ADT1-4 library 1 (human). (C) Schematic diagram of flow sorting of ADT1-4 library 2 (cynomolgus monkey). [Figure 25C] These are schematic diagrams of the selection of mature antibodies using mammalian display. (A) Schematic diagram of flow sorting of the ADT1-7 library. (B) Schematic diagram of flow sorting of ADT1-4 library 1 (human). (C) Schematic diagram of flow sorting of ADT1-4 library 2 (cynomolgus monkey). [Figure 26A]Effect of kappa chain S74L modification on affinity. (A) ADT1-4 strain mammalian display output: Human dissociation off-rate (SPR) vs. human antigen binding (Delfia ELISA, 0.4 nM human antigen) showing the use of LC 74S (white circle) or 74L (black circle). The results highlight the good fit between the affinity improvement by SPR and the affinity improvement by antigen binding by Delfia ELISA. These results also highlight the improved binding conferred by the variable domain S74L modification. (B) ADT1-4 strain mammalian display output: Cynomolgus monkey dissociation off-rate (SPR) vs. cynomolgus monkey antigen binding (Delfia ELISA, 10 nM Cyno antigen) showing the use of LC 74S (white circle) or 74L (black circle). (C) ADT1-4 strain mammalian display output: correlation between improved human antigen dissociation offrate (SPR) versus cynomolgus monkey antigen dissociation offrate (SPR) + LC 74S (white circle) or 74L (black circle). (D) Further confirmation of the effect of S74 vs. L74 on ADT1-4 strain affinity. The kappa chain of SEQ ID NO: 504 was created by mutating the starting serine at position 74 of the kappa chain of the low-affinity parent ADT1-4 to leucine. This modified light chain was paired with the starting ADT1-4 VH (SEQ ID NO: 1) to create the novel molecule "ADT1-4(S74L)" as shown. The kappa chain of SEQ ID NO: 505 was created by changing the starting leucine at position 74 of the kappa chain of the high-affinity ADT1-4-2 back to serine. This was paired with ADT1-4-2 VH (SEQ ID NO: 15) to create the novel molecule "ADT1-4-2(L74S)". The affinity of these molecules (to the starting molecules ADT1-4 and ADT1-4-2, respectively) was determined by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug / mL) was coated onto a Planar Protein A Sensor Chip (Reichert Technologies) to achieve a baseline increase of approximately 500 uRIU.Recombinant human or cynomolgus monkey Vδ1 heterodimers were flowed onto cells at concentrations of 100 nM, 50 nM, 25 nM, 12.5 nM, and 6.25 nM using parameters of association 180 seconds, dissociation 480 seconds, flow rate 25 μL / min, and running buffer PBS + 0.05% Tween 20. All experiments were performed at room temperature: (Di) Results of obtained SPR traces showing antibody affinity for human and cynomolgus monkey Vδ1 antigens. (Dii) Summary of results summarized in the table. [Figure 26B]Effect of kappa chain S74L modification on affinity. (A) ADT1-4 strain mammalian display output: Human dissociation off-rate (SPR) vs. human antigen binding (Delfia ELISA, 0.4 nM human antigen) showing the use of LC 74S (white circle) or 74L (black circle). The results highlight the good fit between the affinity improvement by SPR and the affinity improvement by antigen binding by Delfia ELISA. These results also highlight the improved binding conferred by the variable domain S74L modification. (B) ADT1-4 strain mammalian display output: Cynomolgus monkey dissociation off-rate (SPR) vs. cynomolgus monkey antigen binding (Delfia ELISA, 10 nM Cyno antigen) showing the use of LC 74S (white circle) or 74L (black circle). (C) ADT1-4 strain mammalian display output: correlation between improved human antigen dissociation offrate (SPR) versus cynomolgus monkey antigen dissociation offrate (SPR) + LC 74S (white circle) or 74L (black circle). (D) Further confirmation of the effect of S74 vs. L74 on ADT1-4 strain affinity. The kappa chain of SEQ ID NO: 504 was created by mutating the starting serine at position 74 of the kappa chain of the low-affinity parent ADT1-4 to leucine. This modified light chain was paired with the starting ADT1-4 VH (SEQ ID NO: 1) to create the novel molecule "ADT1-4(S74L)" as shown. The kappa chain of SEQ ID NO: 505 was created by changing the starting leucine at position 74 of the kappa chain of the high-affinity ADT1-4-2 back to serine. This was paired with ADT1-4-2 VH (SEQ ID NO: 15) to create the novel molecule "ADT1-4-2(L74S)". The affinity of these molecules (to the starting molecules ADT1-4 and ADT1-4-2, respectively) was determined by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug / mL) was coated onto a Planar Protein A Sensor Chip (Reichert Technologies) to achieve a baseline increase of approximately 500 uRIU.Recombinant human or cynomolgus monkey Vδ1 heterodimers were flowed onto cells at concentrations of 100 nM, 50 nM, 25 nM, 12.5 nM, and 6.25 nM using parameters of association 180 seconds, dissociation 480 seconds, flow rate 25 μL / min, and running buffer PBS + 0.05% Tween 20. All experiments were performed at room temperature: (Di) Results of obtained SPR traces showing antibody affinity for human and cynomolgus monkey Vδ1 antigens. (Dii) Summary of results summarized in the table. [Figure 26C]Effect of kappa chain S74L modification on affinity. (A) ADT1-4 strain mammalian display output: Human dissociation off-rate (SPR) vs. human antigen binding (Delfia ELISA, 0.4 nM human antigen) showing the use of LC 74S (white circle) or 74L (black circle). The results highlight the good fit between the affinity improvement by SPR and the affinity improvement by antigen binding by Delfia ELISA. These results also highlight the improved binding conferred by the variable domain S74L modification. (B) ADT1-4 strain mammalian display output: Cynomolgus monkey dissociation off-rate (SPR) vs. cynomolgus monkey antigen binding (Delfia ELISA, 10 nM Cyno antigen) showing the use of LC 74S (white circle) or 74L (black circle). (C) ADT1-4 strain mammalian display output: correlation between improved human antigen dissociation offrate (SPR) versus cynomolgus monkey antigen dissociation offrate (SPR) + LC 74S (white circle) or 74L (black circle). (D) Further confirmation of the effect of S74 vs. L74 on ADT1-4 strain affinity. The kappa chain of SEQ ID NO: 504 was created by mutating the starting serine at position 74 of the kappa chain of the low-affinity parent ADT1-4 to leucine. This modified light chain was paired with the starting ADT1-4 VH (SEQ ID NO: 1) to create the novel molecule "ADT1-4(S74L)" as shown. The kappa chain of SEQ ID NO: 505 was created by changing the starting leucine at position 74 of the kappa chain of the high-affinity ADT1-4-2 back to serine. This was paired with ADT1-4-2 VH (SEQ ID NO: 15) to create the novel molecule "ADT1-4-2(L74S)". The affinity of these molecules (to the starting molecules ADT1-4 and ADT1-4-2, respectively) was determined by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug / mL) was coated onto a Planar Protein A Sensor Chip (Reichert Technologies) to achieve a baseline increase of approximately 500 uRIU.Recombinant human or cynomolgus monkey Vδ1 heterodimers were flowed onto cells at concentrations of 100 nM, 50 nM, 25 nM, 12.5 nM, and 6.25 nM using parameters of association 180 seconds, dissociation 480 seconds, flow rate 25 μL / min, and running buffer PBS + 0.05% Tween 20. All experiments were performed at room temperature: (Di) Results of obtained SPR traces showing antibody affinity for human and cynomolgus monkey Vδ1 antigens. (Dii) Summary of results summarized in the table. [Figure 26D]Effect of kappa chain S74L modification on affinity. (A) ADT1-4 strain mammalian display output: Human dissociation off-rate (SPR) vs. human antigen binding (Delfia ELISA, 0.4 nM human antigen) showing the use of LC 74S (white circle) or 74L (black circle). The results highlight the good fit between the affinity improvement by SPR and the affinity improvement by antigen binding by Delfia ELISA. These results also highlight the improved binding conferred by the variable domain S74L modification. (B) ADT1-4 strain mammalian display output: Cynomolgus monkey dissociation off-rate (SPR) vs. cynomolgus monkey antigen binding (Delfia ELISA, 10 nM Cyno antigen) showing the use of LC 74S (white circle) or 74L (black circle). (C) ADT1-4 strain mammalian display output: correlation between improved human antigen dissociation offrate (SPR) versus cynomolgus monkey antigen dissociation offrate (SPR) + LC 74S (white circle) or 74L (black circle). (D) Further confirmation of the effect of S74 vs. L74 on ADT1-4 strain affinity. The kappa chain of SEQ ID NO: 504 was created by mutating the starting serine at position 74 of the kappa chain of the low-affinity parent ADT1-4 to leucine. This modified light chain was paired with the starting ADT1-4 VH (SEQ ID NO: 1) to create the novel molecule "ADT1-4(S74L)" as shown. The kappa chain of SEQ ID NO: 505 was created by changing the starting leucine at position 74 of the kappa chain of the high-affinity ADT1-4-2 back to serine. This was paired with ADT1-4-2 VH (SEQ ID NO: 15) to create the novel molecule "ADT1-4-2(L74S)". The affinity of these molecules (to the starting molecules ADT1-4 and ADT1-4-2, respectively) was determined by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug / mL) was coated onto a Planar Protein A Sensor Chip (Reichert Technologies) to achieve a baseline increase of approximately 500 uRIU.Recombinant human or cynomolgus monkey Vδ1 heterodimers were flowed onto cells at concentrations of 100 nM, 50 nM, 25 nM, 12.5 nM, and 6.25 nM using parameters of association 180 seconds, dissociation 480 seconds, flow rate 25 μL / min, and running buffer PBS + 0.05% Tween 20. All experiments were performed at room temperature: (Di) Results of obtained SPR traces showing antibody affinity for human and cynomolgus monkey Vδ1 antigens. (Dii) Summary of results summarized in the table. [Figure 27A] Use and cross-sharing of CDR3. (A) Exemplary heavy and light chain cross-sharing between affinity-matured antibodies in the ADT1-4(G04) lineage (0.4nM human TRDV1 ELISA yields graded results in the heatmap). This "heatmap" demonstrates the cross-sharing of CDR3 sequences between different affinity-matured antibodies in the ADT1-4 lineage, showing the affinity-mature process that provides antibody sequences that do not need to be provided as specific heavy and light chain pairs. The results highlight that affinity-matured antibodies can cross-share light and heavy chains, and that different LC / HC combinations yield equivalent or improved results in high-stringency antigen-binding studies. The ADT1-4(G04) parental clone is also included in the figure (bottom left). (B) Final selection of the ADT1-4 lineage: Use and cross-sharing of CDR3 + cynomolgus monkey antigen-binding "heatmap" vs. starting parent ADT1-4 parental G04 mAb (bottom left). (C) Final selection of ADT1-7 lineage: CDR3 use and cross-coupling + human antigen binding "heatmap" vs. initiating ADT1-7 parent E07 mAb (bottom left). [Figure 27B]Use and cross-sharing of CDR3. (A) Exemplary heavy and light chain cross-sharing between affinity-matured antibodies in the ADT1-4(G04) lineage (0.4nM human TRDV1 ELISA yields graded results in the heatmap). This "heatmap" demonstrates the cross-sharing of CDR3 sequences between different affinity-matured antibodies in the ADT1-4 lineage, showing the affinity-mature process that provides antibody sequences that do not need to be provided as specific heavy and light chain pairs. The results highlight that affinity-matured antibodies can cross-share light and heavy chains, and that different LC / HC combinations yield equivalent or improved results in high-stringency antigen-binding studies. The ADT1-4(G04) parental clone is also included in the figure (bottom left). (B) Final selection of the ADT1-4 lineage: Use and cross-sharing of CDR3 + cynomolgus monkey antigen-binding "heatmap" vs. starting parent ADT1-4 parental G04 mAb (bottom left). (C) Final selection of ADT1-7 lineage: CDR3 use and cross-coupling + human antigen binding "heatmap" vs. initiating ADT1-7 parent E07 mAb (bottom left). [Figure 27C] Use and cross-sharing of CDR3. (A) Exemplary heavy and light chain cross-sharing between affinity-matured antibodies in the ADT1-4(G04) lineage (0.4nM human TRDV1 ELISA yields graded results in the heatmap). This "heatmap" demonstrates the cross-sharing of CDR3 sequences between different affinity-matured antibodies in the ADT1-4 lineage, showing the affinity-mature process that provides antibody sequences that do not need to be provided as specific heavy and light chain pairs. The results highlight that affinity-matured antibodies can cross-share light and heavy chains, and that different LC / HC combinations yield equivalent or improved results in high-stringency antigen-binding studies. The ADT1-4(G04) parental clone is also included in the figure (bottom left). (B) Final selection of the ADT1-4 lineage: Use and cross-sharing of CDR3 + cynomolgus monkey antigen-binding "heatmap" vs. starting parent ADT1-4 parental G04 mAb (bottom left). (C) Final selection of ADT1-7 lineage: CDR3 use and cross-coupling + human antigen binding "heatmap" vs. initiating ADT1-7 parent E07 mAb (bottom left). [Figure 28]Enhancement factor of binding to ADT1-4 strains compared to ADT1-4 parent G04. (A) Enhancement factor of recombinant human Vδ1 antigen binding. (B) Enhancement factor of recombinant cynomolgus monkey Vδ1 antigen binding. (C) Enhancement factor of primary Vδ1 MFI. (D) Enhancement factor of PEER Vδ1 cell line MFI. [Figure 29] (A) Enhancement factor of binding to ADT1-7 strains compared to ADT1-7 parent E07. (B) Enhancement factor of recombinant human Vδ1 antigen binding. (C) Enhancement factor of primary Vδ1 MFI. [Figure 30A] Improved multiplier of human (and cynomolgus monkey) antigen binding to parental clones. (A) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen (containing human Vδ1 TCR antigen). (B) Improved multiplier of binding to cynomolgus monkey antigens (DV1 / GV77 including cynomolgus monkey SEQ ID NO: 308 (mature, minus leader)). DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen. (C) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-7 (E07) with 0.4 nM recombinant human L1 antigen. [Figure 30B] Improved multiplier of human (and cynomolgus monkey) antigen binding to parental clones. (A) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen (containing human Vδ1 TCR antigen). (B) Improved multiplier of binding to cynomolgus monkey antigens (DV1 / GV77 including cynomolgus monkey SEQ ID NO: 308 (mature, minus leader)). DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen. (C) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-7 (E07) with 0.4 nM recombinant human L1 antigen. [Figure 30C]Improved multiplier of human (and cynomolgus monkey) antigen binding to parental clones. (A) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen (containing human Vδ1 TCR antigen). (B) Improved multiplier of binding to cynomolgus monkey antigens (DV1 / GV77 including cynomolgus monkey SEQ ID NO: 308 (mature, minus leader)). DELFIA ELISA signal of affinity mature clone versus parental ADT1-4 (G04) with 0.4 nM recombinant human L1 antigen. (C) Improved multiplier of binding to human antigens. DELFIA ELISA signal of affinity mature clone versus parental ADT1-7 (E07) with 0.4 nM recombinant human L1 antigen. [Figure 31A] Improvement in the KD factor of human (and cynomolgus monkey) antigens relative to the parent clone. (A) Improvement in the KD factor of human antigens in the ADT1-4(G04) lineage. (B) Improvement in the KD factor of cynomolgus monkey antigens in the ADT1-4(G04) lineage. (C) Improvement in the KD factor of human antigens in the ADT1-7(E07) lineage. [Figure 31B] Improvement in the KD factor of human (and cynomolgus monkey) antigens relative to the parent clone. (A) Improvement in the KD factor of human antigens in the ADT1-4(G04) lineage. (B) Improvement in the KD factor of cynomolgus monkey antigens in the ADT1-4(G04) lineage. (C) Improvement in the KD factor of human antigens in the ADT1-7(E07) lineage. [Figure 31C] Improvement in the KD factor of human (and cynomolgus monkey) antigens relative to the parent clone. (A) Improvement in the KD factor of human antigens in the ADT1-4(G04) lineage. (B) Improvement in the KD factor of cynomolgus monkey antigens in the ADT1-4(G04) lineage. (C) Improvement in the KD factor of human antigens in the ADT1-7(E07) lineage. [Figure 32A] Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 32B] Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 32C] Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 32D] Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 32E] Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 32F]Binding affinity analysis for human Vδ1 antigen (KD by SPR). (A) Surface plasmon resonance of ADT1-4 strains. (B) Surface plasmon resonance of ADT1-7 strains. (C) KD value and multiplicative change relative to parent clone for ADT1-4 strains. (D) KD value and multiplicative change relative to parent clone for ADT1-7 strains. (E) Multiplicative change of KD relative to parent clone for ADT1-4 strains. (F) Multiplicative change of KD relative to parent clone for ADT1-7 strains. [Figure 33] Binding affinity analysis (KD by SPR) for cynomolgus monkey antigen. (A) Surface plasmon resonance of ADT1-4 lines for cynomolgus monkey antigen. (B) KD values ​​of ADT1-4 lines for cynomolgus monkey antigen. [Figure 34A] (A, B) Binding affinity to cell surface Vδ1 TCR (EC50 for binding to cell surface Vδ1). (C) Bar graphs representing mean 50% binding values ​​from ADT1-4 and ADT1-7 clones that bind to Vδ1-positive γδ T cells, represented as an average of two donors. These values ​​are shown in the table along with the % improvement. (D) Table summarizing the IC50 plotted in (A) and (B), as well as Vδ1-negative cell types, including various leukocyte subsets such as HEK293A, Raji cells, and primary blood mononuclear cells. For Vδ1-positive γδ T cells, the data are represented as an average of two donors. [Figure 34B] (A, B) Binding affinity to cell surface Vδ1 TCR (EC50 for binding to cell surface Vδ1). (C) Bar graphs representing mean 50% binding values ​​from ADT1-4 and ADT1-7 clones that bind to Vδ1-positive γδ T cells, represented as an average of two donors. These values ​​are shown in the table along with the % improvement. (D) Table summarizing the IC50 plotted in (A) and (B), as well as Vδ1-negative cell types, including various leukocyte subsets such as HEK293A, Raji cells, and primary blood mononuclear cells. For Vδ1-positive γδ T cells, the data are represented as an average of two donors. [Figure 34C] (A, B) Binding affinity to cell surface Vδ1 TCR (EC50 for binding to cell surface Vδ1). (C) Bar graphs representing mean 50% binding values ​​from ADT1-4 and ADT1-7 clones that bind to Vδ1-positive γδ T cells, represented as an average of two donors. These values ​​are shown in the table along with the % improvement. (D) Table summarizing the IC50 plotted in (A) and (B), as well as Vδ1-negative cell types, including various leukocyte subsets such as HEK293A, Raji cells, and primary blood mononuclear cells. For Vδ1-positive γδ T cells, the data are represented as an average of two donors. [Figure 34D] (A, B) Binding affinity to cell surface Vδ1 TCR (EC50 for binding to cell surface Vδ1). (C) Bar graphs representing mean 50% binding values ​​from ADT1-4 and ADT1-7 clones that bind to Vδ1-positive γδ T cells, represented as an average of two donors. These values ​​are shown in the table along with the % improvement. (D) Table summarizing the IC50 plotted in (A) and (B), as well as Vδ1-negative cell types, including various leukocyte subsets such as HEK293A, Raji cells, and primary blood mononuclear cells. For Vδ1-positive γδ T cells, the data are represented as an average of two donors. [Figure 35A] TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35B]TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35C] TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35D] TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35E]TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35F] TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 35G] TCR downregulation (by soluble antibodies). (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Mean TCR downregulation - IC50 obtained from two GD cell donors. (D, E) Improved polynomial TCR downregulation from parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). (G) 50% effect value from (C) calculated from each parent of ADT1-4 (top) and ADT1-7 (bottom). [Figure 36A]Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 36B] Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 36C] Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 36D]Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 36E] Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 36F] Vδ1 monoclonal antibody against γδ activation as measured by CD107a expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD107a expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37A]Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37B] Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37C] Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37D]Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37E] Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 37F] Vδ1 monoclonal antibody against γδ activation as measured by CD25 expression. (A, B) GD cells alone (A) or ADT1-4-2 clone with THP-1 cells (B). (C, D) ADT1-7-3 clone in GD cells alone (C) or with THP-1 cells (D). (E) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated non-co-cultured γδ cells. (F) Table showing the percentage increase in γδCD25 expression from co-cultured cells treated with the highest concentration of Vδ1 mAb compared to untreated, co-cultured, and non-co-cultured γδ cells. [Figure 38A]Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38B] Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38C] Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38D] Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38E]Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38F] Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 38G] Downregulation of TCR in cynomolgus monkeys (by soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to associate with VD1 on cynomolgus monkey γδ-T cells and reduce its expression. (B) Percentage of VD1 cell surface expression after treatment with ADT1-4-2. (C) Individual EC50 values ​​of different donors combined with mean and standard deviation. (D-G) Additional tests performed using whole blood samples from cynomolgus monkeys (n=5). [Figure 39A] Quantification of the number of viable THP-1 cells after co-culture with γδT cells for 24 hours in the presence of Vδ1 mAb or control. (A) THP-1 cytotoxicity assay for ADT1-4 clones. (B) THP-1 cytotoxicity assay for ADT1-7 clones. (C) Mean EC50 in the THP-1 cytotoxicity assays for ADT1-4 and ADT1-7 clones. (D) Table summarizing the EC50s plotted in (C). [Figure 39B]Quantification of the number of viable THP-1 cells after co-culture with γδT cells for 24 hours in the presence of Vδ1 mAb or control. (A) THP-1 cytotoxicity assay for ADT1-4 clones. (B) THP-1 cytotoxicity assay for ADT1-7 clones. (C) Mean EC50 in the THP-1 cytotoxicity assays for ADT1-4 and ADT1-7 clones. (D) Table summarizing the EC50s plotted in (C). [Figure 39C] Quantification of the number of viable THP-1 cells after co-culture with γδT cells for 24 hours in the presence of Vδ1 mAb or control. (A) THP-1 cytotoxicity assay for ADT1-4 clones. (B) THP-1 cytotoxicity assay for ADT1-7 clones. (C) Mean EC50 in the THP-1 cytotoxicity assays for ADT1-4 and ADT1-7 clones. (D) Table summarizing the EC50s plotted in (C). [Figure 39D] Quantification of the number of viable THP-1 cells after co-culture with γδT cells for 24 hours in the presence of Vδ1 mAb or control. (A) THP-1 cytotoxicity assay for ADT1-4 clones. (B) THP-1 cytotoxicity assay for ADT1-7 clones. (C) Mean EC50 in the THP-1 cytotoxicity assays for ADT1-4 and ADT1-7 clones. (D) Table summarizing the EC50s plotted in (C). [Figure 40] A Vδ1 monoclonal antibody against antibody-dependent cell-mediated cytotoxicity (ADCC). [Figure 41] A Vδ1 monoclonal antibody against complement-dependent cell-mediated cytotoxicity (CDC). [Figure 42] Preservation of healthy cells in clone ADT1-4-2. [Figure 43A]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43B]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43C]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43D]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43E]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43F]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43G]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43H]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43I]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 43J]Effects of anti-Vδ1 antibodies on a TIL population derived from primary tumor biopsy. (A, B) Shows the percentage decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 48(A) or 72(B) hours of mAb stimulation in two separate donors, with target association confirmed in each case. (C) Enhanced expression of both CD25 and Ki67 on Vδ1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or stimulation with ADT1-4. (D) Shows a substantial multiplicative increase in IFN-γ production by TILs stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50 ng / ml IL-15 for 72 hours. (E) Shows that stimulation of TILs with ADT-1-4-2 or ADT1-7-3 did not enhance the secretion of type 17-related cytokines IL-6 or IL-17 at this point. (F, G) Shows a decrease in Vδ1 TCR expression on total tumor-infiltrating γδ T cells after 24(F) or 72(G) hours of mAb stimulation in two individual donors, confirming target association in TILs isolated by enzymatic digestion. (H) Shows dose-dependent enhancement of Ki67 expression on γδ T cells after 72 hours of stimulation with ADT1-4-2. (I, J) Shows a multiplicative increase in IFN-γ produced by TILs isolated from two individual donors by enzymatic digestion and stimulated with a concentration of 6.66 nM ADT1-4-2 in the presence of 2 ng / ml IL-15 for 24(I) or 72(J). In all cases, the concentrations of control anti-RSV and parental ADT1-4 are consistent with the highest concentrations used in the study (i.e., concentrations of 6.66 nM for A, C, F, G, H, I, and J, and 66.6 nM for B and E). [Figure 44]Effects of anti-Vδ1 antibody on TIL population from primary tumor biopsy. (A) Shows enhanced expression of CD25 and Ki67 in γδ T cells stimulated with ADT1-4-2 for 10 days. (B) Shows a substantial increase in perforin + granzyme B + γδ T cells after 10 days of stimulation with ADT1-4-2. (C) Shows a considerable enhancement of granzyme B and perforin expression in both CD8+ and CD8-αβ T cells after 10 days of stimulation of tumor-infiltrating Vδ1+ T cells with ADT1-4-2. (D) Shows a marked increase in IFN-γ production and a moderate increase in IL-17 and IL-6 production by lung tumor-derived TILs after stimulation with ADT1-4-2. I demonstrates enhanced production of chemokines CCL2, CCL4 and CXCL10 by TILs after 10 days of stimulation with ADT1-4-2. In all cases, the control anti-RSV and parental ADT1-4 were matched in concentration to affinity-mature ADT1-4-2. [Figure 45] Sequence of ADT1-4 lineage clones (light chain) [Figure 46] Sequence of ADT1-4 lineage clones (heavy chain). [Figure 47] Sequence of ADT1-7 lineage clones (light chain). [Figure 48] Sequence of ADT1-7 lineage clones (heavy chain). [Figure 49] The anti-Vδ1 / CD19 bispecific antibody exhibits high affinity binding to human Vδ1 and cynomolgus monkey Vδ1, comparable to that of the parental monoclonal Vδ1 mAb. Surface plasmon resonance (SPR) analysis using the ADT1-4-2 / CD19 bispecific agent was performed to evaluate binding to human Vδ1 and CD19 antigens. The bispecific antibody bound to both human and cynomolgus monkey Vδ1, with only minimal reduction in affinity for both antigens. [Figure 50A]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50B]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50C]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50D]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50E]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50F]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50G]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50H]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50I]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50J]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 50K]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. (A) CD19 expression on cancerous NALM-6 and Raji cells, as well as primary B cells and Vδ1γδ T cells, as determined by flow cytometry. (B-D) Effects of parental (ADT1-4) and affinity-matured (ADT1-4-2) Vδ1-CD19 bispecificity on NALM-6 cells (B), Raji cells (C), and B cell cytotoxicity (D). Antibodies were titrated over 12 hours in the presence of Vδ1γδ T cells with a 1:1 E:T ratio. The percentage of viable cells was calculated by high-concentration confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδ T cells. (E)(B), (C), and (D) are bar graphs showing the percentage of viable cells after 12 hours of co-culture of Vδ1γδT cells with either NALM-6, Raji, or B cells in the presence of Vδ1 bispecificity and a control. (F~G) Effects of Vδ1-CD19 bispecificity on Vδ1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G), as determined by flow cytometry after 4 hours of co-culture. (H) Bar graphs showing the maximum percentage of TCR downregulation as shown in (F) and (G) at the highest concentration (3 μg / ml). (I~K) Effects of Vδ1-CD19 bispecificity on CD107a upregulation on the surface of Vδ1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J), as determined by flow cytometry after 4 hours of co-culture. All data are shown mean ± standard deviation and are representative of n=3. Blinatumomab (CD3-CD19 bispecific (BiTE)) was included as a control in all assays. [Figure 51A]Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51B] Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51C]Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51D] Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51E]Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51F] Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51G]Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51H] Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51I]Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 51J] Bispecificity-format affinity-matured Vδ1 clones bind to Her2+ target cells, exhibiting enhanced binding to Vδ1γδT cells and enhanced cytotoxicity of Her2+ target cells. (A) Cell surface expression of Her2 and Vδ1 on breast cancer cell lines and Vδ1γδT cells (B). (C~F) Conjugation of Vδ1-Her2 bispecific antibody and Her2 mAb control (trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), Her2- (MDA-MD-231 (E)), and Vδ1+ cells (F). (G~I) Percentage of viable cells remaining after 24 hours of co-culture of Vδ1γδT cells with SK-BR-3 cells (G), BT-474 (H), and MDA-MB-231 cells (I) in a 1:1 E:T ratio in the presence of Vδ1-Her2 bispecific antibody. (J) Bar graph showing the percentage increase in cytotoxicity of Vδ1γδT cells with Vδ1-Her2 bispecific antibody after 24 hours. [Figure 52A]The Vδ1 / EGFR bispecific antibody exhibits high affinity binding to human EGFR and human Vδ1 binding affinity comparable to its parent mAb. (A, B) Surface plasmon resonance (SPR) analysis was performed using Vδ1 / EGFR bispecific mutants to evaluate binding to human Vδ1 (A) and human EGFR antigen (B). For comparison, the parent mAb, cetuximab, and a negative control mAb were included. [Figure 52B] The Vδ1 / EGFR bispecific antibody exhibits high affinity binding to human EGFR and human Vδ1 binding affinity comparable to its parent mAb. (A, B) Surface plasmon resonance (SPR) analysis was performed using Vδ1 / EGFR bispecific mutants to evaluate binding to human Vδ1 (A) and human EGFR antigen (B). For comparison, the parent mAb, cetuximab, and a negative control mAb were included. [Figure 53A] The Vδ1 / EGFR bispecific antibody binds to EGFR+A431 target cells and Vδ1γδT cells. (A) Cell surface expression of EGFR and Vδ1 on A431 cell lines and primary Vδ1γδT cells. (B) Binding level of the Vδ1 / EGFR bispecific antibody to A431 cell lines or primary Vδ1γδT cells. Target cells were stained with various concentrations of antibody and then stained with a fluorescent anti-human IgG detection antibody. All incubation steps were performed at 4°C, and mAb binding was determined by measuring the median fluorescence level using flow cytometry. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. [Figure 53B] The Vδ1 / EGFR bispecific antibody binds to EGFR+A431 target cells and Vδ1γδT cells. (A) Cell surface expression of EGFR and Vδ1 on A431 cell lines and primary Vδ1γδT cells. (B) Binding level of the Vδ1 / EGFR bispecific antibody to A431 cell lines or primary Vδ1γδT cells. Target cells were stained with various concentrations of antibody and then stained with a fluorescent anti-human IgG detection antibody. All incubation steps were performed at 4°C, and mAb binding was determined by measuring the median fluorescence level using flow cytometry. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. [Figure 54A] The Vδ1 / EGFR bispecific antibody induces activation and degranulation of EGFR-specific T cells, resulting in increased γδ T cell-mediated cytotoxicity of A431 target cells. (A) Cell surface expression of γδ TCR in primary Vδ1γδ T cells after 24 hours of culture with the bispecific antibody in or without A431 cells. (B, C) Number of viable A431 cells (B) and activation status of primary Vδ1γδ T cells after 24 hours of co-culture with various concentrations of antibody in a 1:1 ratio. Viability was measured using a viability-determinating dye, and activation status was measured using CD25 antibody. (D) Degranulation of primary Vδ1γδ T cells after 4 hours of co-culture with A431 cells in a 1:1 ratio with various concentrations of antibody. Degranulation was determined by directly adding anti-CD107α antibody conjugated to a fluorophore to the cell-antibody mix at the start of co-culture. (E) Number of viable A431 cells after 24 hours co-culture with 10 pM antibody and varying amounts of primary Vδ1γδT cells. (A-E) In all cases, fluorescence was determined using flow cytometry to measure the median fluorescence level. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. Data are expressed as mean ± SD of two biological replicas. [Figure 54B]The Vδ1 / EGFR bispecific antibody induces activation and degranulation of EGFR-specific T cells, resulting in increased γδ T cell-mediated cytotoxicity of A431 target cells. (A) Cell surface expression of γδ TCR in primary Vδ1γδ T cells after 24 hours of culture with the bispecific antibody in or without A431 cells. (B, C) Number of viable A431 cells (B) and activation status of primary Vδ1γδ T cells after 24 hours of co-culture with various concentrations of antibody in a 1:1 ratio. Viability was measured using a viability-determinating dye, and activation status was measured using CD25 antibody. (D) Degranulation of primary Vδ1γδ T cells after 4 hours of co-culture with A431 cells in a 1:1 ratio with various concentrations of antibody. Degranulation was determined by directly adding anti-CD107α antibody conjugated to a fluorophore to the cell-antibody mix at the start of co-culture. (E) Number of viable A431 cells after 24 hours co-culture with 10 pM antibody and varying amounts of primary Vδ1γδT cells. (A-E) In all cases, fluorescence was determined using flow cytometry to measure the median fluorescence level. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. Data are expressed as mean ± SD of two biological replicas. [Figure 54C]The Vδ1 / EGFR bispecific antibody induces activation and degranulation of EGFR-specific T cells, resulting in increased γδ T cell-mediated cytotoxicity of A431 target cells. (A) Cell surface expression of γδ TCR in primary Vδ1γδ T cells after 24 hours of culture with the bispecific antibody in or without A431 cells. (B, C) Number of viable A431 cells (B) and activation status of primary Vδ1γδ T cells after 24 hours of co-culture with various concentrations of antibody in a 1:1 ratio. Viability was measured using a viability-determinating dye, and activation status was measured using CD25 antibody. (D) Degranulation of primary Vδ1γδ T cells after 4 hours of co-culture with A431 cells in a 1:1 ratio with various concentrations of antibody. Degranulation was determined by directly adding anti-CD107α antibody conjugated to a fluorophore to the cell-antibody mix at the start of co-culture. (E) Number of viable A431 cells after 24 hours co-culture with 10 pM antibody and varying amounts of primary Vδ1γδT cells. (A-E) In all cases, fluorescence was determined using flow cytometry to measure the median fluorescence level. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. Data are expressed as mean ± SD of two biological replicas. [Figure 54D]The Vδ1 / EGFR bispecific antibody induces activation and degranulation of EGFR-specific T cells, resulting in increased γδ T cell-mediated cytotoxicity of A431 target cells. (A) Cell surface expression of γδ TCR in primary Vδ1γδ T cells after 24 hours of culture with the bispecific antibody in or without A431 cells. (B, C) Number of viable A431 cells (B) and activation status of primary Vδ1γδ T cells after 24 hours of co-culture with various concentrations of antibody in a 1:1 ratio. Viability was measured using a viability-determinating dye, and activation status was measured using CD25 antibody. (D) Degranulation of primary Vδ1γδ T cells after 4 hours of co-culture with A431 cells in a 1:1 ratio with various concentrations of antibody. Degranulation was determined by directly adding anti-CD107α antibody conjugated to a fluorophore to the cell-antibody mix at the start of co-culture. (E) Number of viable A431 cells after 24 hours co-culture with 10 pM antibody and varying amounts of primary Vδ1γδT cells. (A-E) In all cases, fluorescence was determined using flow cytometry to measure the median fluorescence level. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. Data are expressed as mean ± SD of two biological replicas. [Figure 54E]The Vδ1 / EGFR bispecific antibody induces activation and degranulation of EGFR-specific T cells, resulting in increased γδ T cell-mediated cytotoxicity of A431 target cells. (A) Cell surface expression of γδ TCR in primary Vδ1γδ T cells after 24 hours of culture with the bispecific antibody in or without A431 cells. (B, C) Number of viable A431 cells (B) and activation status of primary Vδ1γδ T cells after 24 hours of co-culture with various concentrations of antibody in a 1:1 ratio. Viability was measured using a viability-determinating dye, and activation status was measured using CD25 antibody. (D) Degranulation of primary Vδ1γδ T cells after 4 hours of co-culture with A431 cells in a 1:1 ratio with various concentrations of antibody. Degranulation was determined by directly adding anti-CD107α antibody conjugated to a fluorophore to the cell-antibody mix at the start of co-culture. (E) Number of viable A431 cells after 24 hours co-culture with 10 pM antibody and varying amounts of primary Vδ1γδT cells. (A-E) In all cases, fluorescence was determined using flow cytometry to measure the median fluorescence level. Log-scale 4-parameter dose-response curve fitting was performed using GraphPad Prism 9. Data are expressed as mean ± SD of two biological replicas. [Figure 55A] Further evidence of non-depletion and CD3 downregulation in both blood-derived and tumor-associated Vδ1 T cells is presented. (A) Vδ1 TCR MFI upon antibody stimulation is shown as an indicator of mAb target association to blood-derived Vδ1 T cells. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. Stimulation with the vδ1 antibody resulted in association with vδ1 cells and downregulation of both vδ1 and CD3 on vδ1 cells. (C, D, and E) The conferred activation and non-depletion effect on tumor-associated Vδ1 T cells by exemplary anti-Vδ1 antibodies of the present invention is shown. [Figure 55B]Further evidence of non-depletion and CD3 downregulation in both blood-derived and tumor-associated Vδ1 T cells is presented. (A) Vδ1 TCR MFI upon antibody stimulation is shown as an indicator of mAb target association to blood-derived Vδ1 T cells. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. Stimulation with the vδ1 antibody resulted in association with vδ1 cells and downregulation of both vδ1 and CD3 on vδ1 cells. (C, D, and E) The conferred activation and non-depletion effect on tumor-associated Vδ1 T cells by exemplary anti-Vδ1 antibodies of the present invention is shown. [Figure 55C] Further evidence of non-depletion and CD3 downregulation in both blood-derived and tumor-associated Vδ1 T cells is presented. (A) Vδ1 TCR MFI upon antibody stimulation is shown as an indicator of mAb target association to blood-derived Vδ1 T cells. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. Stimulation with the vδ1 antibody resulted in association with vδ1 cells and downregulation of both vδ1 and CD3 on vδ1 cells. (C, D, and E) The conferred activation and non-depletion effect on tumor-associated Vδ1 T cells by exemplary anti-Vδ1 antibodies of the present invention is shown. [Figure 55D] Further evidence of non-depletion and CD3 downregulation in both blood-derived and tumor-associated Vδ1 T cells is presented. (A) Vδ1 TCR MFI upon antibody stimulation is shown as an indicator of mAb target association to blood-derived Vδ1 T cells. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. Stimulation with the vδ1 antibody resulted in association with vδ1 cells and downregulation of both vδ1 and CD3 on vδ1 cells. (C, D, and E) The conferred activation and non-depletion effect on tumor-associated Vδ1 T cells by exemplary anti-Vδ1 antibodies of the present invention is shown. [Figure 55E]Further evidence of non-depletion and CD3 downregulation in both blood-derived and tumor-associated Vδ1 T cells is presented. (A) Vδ1 TCR MFI upon antibody stimulation is shown as an indicator of mAb target association to blood-derived Vδ1 T cells. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. Stimulation with the vδ1 antibody resulted in association with vδ1 cells and downregulation of both vδ1 and CD3 on vδ1 cells. (C, D, and E) The conferred activation and non-depletion effect on tumor-associated Vδ1 T cells by exemplary anti-Vδ1 antibodies of the present invention is shown. [Figure 56A] The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56B]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56C]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56D]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56E]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56F]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56G]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 56H]The Vδ1×FAPα bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of FAPα+ fibroblasts. (A) Binding kinetics of the anti-Vδ1(ADT-1-4-2), anti-FAPα monoclonal (cibrotuzumab-based), and anti-Vδ1xFAPα(ADT-1-4-2xcibrotuzumab) bispecific antibodies to recombinant human Vδ1 and FAPα as measured by surface plasmon resonance (SPR). (B, C) Binding of the anti-Vδ1 antibody and anti-FAPα antibody to FAPα+ fibroblasts (B) and Vδ1γδT cells (C). (D, E) Effects of the anti-Vδ1×FAPα bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of FAPα+ fibroblasts. (F~G) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of FAPα+ fibroblasts. (H) Effects of anti-Vδ1×FAPα bispecific antibody and monoclonal control on the lysis of fibroblasts by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57A]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57B]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57C]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57D]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57E]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57F]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57G]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 57H]The Vδ1×MSLN bispecific antibody enhances the activation of Vδ1γδT cells and the lysis of MSLN+ target cells. (A) Binding kinetics of anti-Vδ1(ADT-1-4-2), anti-MSLN monoclonal (based on the antibody disclosed in U.S. Patent Application Publication No. 2014 / 0004121), and anti-Vδ1×MSLN(ADT1-4-2xMSLN) bispecific antibodies to recombinant human Vδ1 and MSLN, as determined by surface plasmon resonance (SPR). (B, C) Binding of anti-Vδ1 and anti-MSLN antibodies to MSLN+ HeLa cells (B) and Vδ1γδT cells (C). (D, E) Effects of anti-Vδ1×MSLN bispecific and monoclonal antibodies on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (D) or presence (E) of MSLN+OVCAR-3 cells. (F, G) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the upregulation of CD107a in Vδ1γδT cells in the absence (F) or presence (G) of MSLN+OVCAR-3 cells. (H) Effects of anti-Vδ1×MSLN bispecific antibody and monoclonal control on the lysis of HeLa cells by Vδ1γδT cells after 24 hours of co-culture. Cell viability was calculated by high-content confocal microscopy and normalized to the number of viable cells in the absence of Vδ1γδT cells. [Figure 58A]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58B]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58C]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58D]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58E]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58F]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 58G]The Vδ1×PD-1 bispecific antibody enhances the activation of Vδ1γδT cells and inhibits checkpoint blockade of PD-1+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-PD-1(pembrolizumab-based), anti-RSVIgG control×anti-PD-1, and anti-Vδ1×anti-PD-1(ADT1-4-2×pembrolizumab) bispecific antibodies binding to recombinant human Vδ1 and PD-1. B) Double binding of anti-Vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and Vδ1 as determined by SPR. C) Expression of PD-1 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD8 T cells and Vδ1γδT cells. E) Effects of anti-Vδ1×PD-1 bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of PD-1+CD4 T cells. F) EC50 of anti-Vδ1×PD-1 bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of PD-1+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×PD-1 bispecific antibody on PD-1+ T cell activation. [Figure 59A]The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59B] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59C] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59D] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59E] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59F] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 59G] The Vδ1×4-1BB bispecific antibody enhances the activation of Vδ1γδT cells and 4-1BB+ T cells. A) SPR analysis of anti-Vδ1 (ADT1-4-2), anti-4-1BB (utomirumab-based), anti-RSVIgG control × anti-4-1BB (ADT1-4-2 × utomirumab), and anti-Vδ1×anti-4-1BB bispecific antibodies binding to recombinant human Vδ1 and 4-1BB. B) Double binding of anti-Vδ1 and anti-4-1BB bispecific antibodies to recombinant human 4-1BB and Vδ1 as determined by SPR. C) Expression of 4-1BB in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-4-1BB antibodies to 4-1BB+ activated CD8 T cells and Vδ1γδT cells. E-F) Effects of anti-Vδ1×4-1BB bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of 4-1BB+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×4-1BB bispecific antibody on the activation of 4-1BB+ T cells. [Figure 60A] The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60B] The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60C] The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60D]The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60E] The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60F]The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 60G] The Vδ1×OX40 bispecific antibody enhances the activation of Vδ1γδ T cells and OX40+ T cells. A) SPR analysis of anti-Vδ1(ADT1-4-2), anti-OX40(pogalizumab-based), anti-RSVIgG control×anti-OX40, and anti-Vδ1×anti-OX40(ADT1-4-2×pogalizumab) bispecific antibodies binding to recombinant human Vδ1 and OX40. B) Double binding of anti-Vδ1 and anti-OX40 bispecific antibodies to recombinant human OX40 and Vδ1 as determined by SPR. C) Expression of OX40 in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-OX40 antibodies to OX40+ activated CD4 T cells and Vδ1γδ T cells. E-F) Effects of anti-Vδ1×OX40 bispecific antibodies and monoclonal controls on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence (E) or presence (F) of OX40+CD4 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×OX40 bispecific antibodies on the activation of OX40+ T cells. [Figure 61A]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61B]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61C]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61D]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61E]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61F]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 61G]The Vδ1 x TIGIT bispecific antibody enhances the activation of Vδ1γδ T cells and inhibits checkpoint blockade of TIGIT+ T cells. A) SPR analysis of anti-Vδ1(ADT-1-4-2), anti-TIGIT(tiragolumab-based), anti-RSVIgG control x anti-TIGIT, and anti-Vδ1 x anti-TIGIT(ADT-1-4-2 x tiragolumab) bispecific antibodies binding to recombinant human Vδ1 and TIGIT. B) Double binding of anti-Vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and Vδ1 as determined by SPR. C) TIGIT expression in CD4 and CD8 T cells activated with anti-CD3 / anti-CD28 Dynabeads. D) Binding of anti-Vδ1 and anti-TIGIT antibodies to TIGIT+ activated CD4 and CD8 T cells and Vδ1γδ T cells. E) Effects of anti-Vδ1×TIGIT bispecific antibody and monoclonal control on the downregulation of Vδ1 TCR in Vδ1γδT cells in the absence or presence of TIGIT+CD8 T cells. F) EC50 of anti-Vδ1×TIGIT bispecific antibody on the downregulation of Vδ1 TCR in Vδ1γδT cells in the presence or absence of TIGIT+CD8 T cells. G) Effects of Vδ1 crosslinked anti-Vδ1×TIGIT bispecific antibody on TIGIT+ T cell activation. [Figure 62] The ADCC reporter bioassay does not show ADCC as a result of anti-vδ1 antibody. Target cells, i.e., γδ cells, were incubated with ADCC bioassay effector cells in the presence of anti-vδ1 antibody, anti-vδ1 LAGA antibody (Fc inactive), and RSV isotype control. Luminescence signals were recorded as relative light units (RLU), and induction magnification was calculated as described in the methods. For "anti-vδ1 antibody," "anti-vδ1 LAGA antibody," "RSV," and "OKT3," N=2 γδ donors were used (technical duplicate). For the "rituximab + Raji" condition (technical duplicate), N=1 Raji cell line was used. For the "anti-vδ1 antibody + effector" and "anti-vδ1 LAGA antibody + effector" conditions (technical duplicate and single-dose, respectively), n=1 γδ donor was used. The effector:target ratio was 3:1. [Figure 63A] The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63B]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63C]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63D]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63E]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63F]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63G]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63H]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 63I]The Vδ1-CD19 bispecific T cell engager enhances CD19+ target cytotoxicity and γδ T cell activation while preserving healthy CD19+ B cells. A-F) Effects of anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies on γδ T cell or αβ T cell-mediated lysis of Raji cells or healthy primary B cells. The positivity of Raji cells or healthy primary B cells was determined at 24 hours by confocal microscopy in three cultures: Vδ1γδ T cells, Raji cells (A), and healthy primary B cells (B); three cultures: αβ T cells, Raji cells (C), and healthy primary B cells (D); or four cultures: Vδ1γδ T cells, αβ T cells, Raji cells (E), and healthy primary B cells (F). G-I) Quantification of IL-17A secretion from γδT cells or αβT cells 24 hours after stimulation with anti-Vδ1×CD19 and CD3×CD19 bispecific antibodies. Cell culture supernatants were collected from Raji cells, primary B cells, and co-cultures of γδT cells (G); or αβT cells (H); or γδT cells and αβT cells (I), and IL-17A secretion was determined by MSD. The dotted line represents the lowest level of quantification. [Figure 64]Stimulation with a high-affinity anti-Vδ1+ antibody enhances 4-1BB expression on Vδ1γδT cells. (A) Exemplary enhancement of 4-1BB expression by addition of a high-affinity anti-Vδ1 antibody: Vδ1+ T cells derived from tissues of four different human donors were incubated with the indicated antibody (1 μg / ml) in the presence of THP-1 cells (2:1 E:T). After 4 hours, 4-1BB expression on the surface of Vδ1+ T cells was analyzed by flow cytometry. Cells stimulated with ADT1-4-2 showed a significant increase in 4-1BB expression at this point, which was comparable to stimulation with OKT3 (*p<0.05, **p<0.01 by standard one-way ANOVA with Sidak's post-hoc test). (B, C) Enhanced 4-1BB Expression; Further Tests: Tissue-derived Vδ1+ T cells were incubated with escalating doses of ADT1-4-2 or an anti-IgG1 isotype control, as shown in a 1:1 ratio in either (B) THP-1 cancer cells or (C) healthy blood-derived monocytes. After 4 hours, 4-1BB expression on the surface of Vδ1+ T cells was analyzed by flow cytometry. Stimulation with ADT1-4-2 increased 4-1BB expression on the surface of Vδ1 T cells. Furthermore, similar antibody-induced activation was observed when crosslinking was provided by tumor or healthy FcγR+ cells, even if the activated Vδ1+ cells preserved healthy cells (see other parts of this specification for further discussion). [Figure 65]Experimental culture setup including analysis of innate cytotoxic receptors (NCR upregulation (A) and TCR downregulation (A)) on Vδ1+ T cells in primary PBMCs: Human PBMC cultures from three different donors were incubated for 10 days with exemplary affinity matured ADT1-4 and ADT1-7 anti-Vδ1 antibody clones (see also B, C, and D) at 3.33 nM as shown, or with anti-RSV controls. Cultures were also supplemented with 4 ng / ml IL-2 and 10 ng / ml IL-15 (Miltenyi, 130-097-746, and Peprotech, 200-15-500 ug, respectively). After 10 days of incubation, % Vδ1+ T cells were analyzed by flow cytometry. All cultures incubated with anti-Vδ1 antibodies showed a significant decrease in detectable Vδ1+ cells (as % panγ) compared to anti-RSV control cultures. Representative and typical results for one such antibody are shown here. Such analysis makes it possible to confirm that all affinity-mature anti-Vδ1 antibodies included in this study showed significant target association and resulted in downregulation of TCR. (B~D) Anti-Vδ1 antibodies resulted in enhanced expression of NCRs (NKp30, NKp44, NKp46) on detectable Vδ1+ cells: In addition to confirming target association for all anti-Vδ1 antibodies used in this study, on gated Vδ1+ cells... The multiplicative changes in NCR were also tested. Specifically, (B) shows the multiplicative changes of double-positive Vδ1+ and NKp 30+% relative to the RSV control (normalized to the right and represented as 1x). (C) shows the multiplicative changes of double-positive Vδ1+ and NKp 44+% relative to the RSV control (normalized to the right and represented as 1x). (D) shows the multiplicative changes of double-positive Vδ1+ and NKp 46+% relative to the RSV control (normalized to the right and represented as 1x). These combined results highlight the consistent (donor-dependent) upregulation of NCR in detectable Vδ1+ conferred by all affinity-matured anti-Vδ1+ antibodies used in this extended 10-day primary PBMC culture system. [Modes for carrying out the invention]

[0043] The present invention provides high-affinity anti-TCR delta variable region 1 (anti-Vδ1) antibodies, multispecific antibodies, and antibody fragments thereof. More specifically, the present invention relates to the provision and characterization of antibodies prepared by an optimized selection procedure starting from parental anti-Vδ1 antibodies, such as optimized antibodies, for example the parental antibodies referred to herein as G04, E07, C08, B07, C05, E04, F07, G06, G09, B09, G10, and E01. In particular, the present invention relates to optimized antibodies derived from G04 and E07.

[0044] Definitions Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which the present invention pertains. As used herein, the following terms shall have the meanings set forth below as assigned to them.

[0045] Gamma-delta (γδ) T cells represent a small subset of T cells that express a distinct, characteristic T cell receptor (TCR) on their surface. This TCR is composed of one gamma (γ) chain and one delta (δ) chain. Each chain includes 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 non-hematopoietic tissues. The two subtypes can be defined by the type of δ and / or γ present in the cells. For example, γδ T cells that predominate in peripheral blood predominantly express the delta variable region 2 chain (Vδ2). γδ T cells that predominate in non-hematopoietic tissues (i.e., are tissue-resident) predominantly express the delta variable region 1 chain. Reference to "Vδ1 T cells" or "Vδ1+ T cells" refers to γδ T cells having a Vδ1 chain, that is, Vδ1+ cells.

[0046] Reference to "delta variable region 1" may also be referred to as Vδ1 or Vd1, while a nucleotide encoding a TCR chain comprising this region, or a TCR protein complex comprising this region, may be referred to as "TRDV1". All antibodies or antigen-binding fragments thereof that interact with the Vδ1 chain of a γδ TCR are, in effect, antibodies or antigen-binding fragments thereof that bind to Vδ1, and may be referred to as "anti-TCR delta variable region 1 antibody or antigen-binding fragment thereof" or "anti-Vδ1 antibody or antigen-binding fragment thereof" or "anti-TRDV1 antibody or antigen-binding fragment thereof".

[0047] Further reference is made herein to other delta chains, such as the "delta variable region 2" chain. These may be referred to in a similar manner. For example, the delta variable region 2 chain may be referred to as Vδ2, while a nucleotide encoding a TCR chain comprising this region, or a TCR protein complex comprising this region, may be referred to as "TRDV2". In a preferred embodiment, the antibody or antigen-binding fragment thereof that interacts with the Vδ1 chain of a γδ TCR does not interact with other delta chains such as Vδ2. In the present invention, the antibody is specific for TRDV1 and does not bind to TRDV2 (SEQ ID NO: 310) or other antigens present on γδ T cell receptors, for example, TRDV3 (SEQ ID NO: 311).

[0048] Reference is also made herein to "gamma variable chains". While these may be referred to as gamma chains or Vγ, a nucleotide encoding a TCR chain comprising this region, or a TCR protein complex comprising this region, may be referred to as TRGV. For example, TRGV4 refers to the Vγ4 chain. In a preferred embodiment, the antibody or antigen-binding fragment thereof that interacts with the Vδ1 chain of a γδ TCR does not interact with gamma chains such as Vγ4 (e.g., SEQ ID NO: 309). In a preferred embodiment, the antibody also does not bind or interact with other domains found within γδ TCRs, such as TRDJ, TRDC, TRGJ, or TRGC.

[0049] The term "T cell receptor complex" refers to a complex of proteins containing the "T cell receptor" (or "TCR") found on the surface of T cells, which are responsible for recognizing various antigens. The T cell receptor complex may contain the α and β chains of the T cell receptor, or, in the case of γδ T cells, the γ and δ chains of the T cell receptor, as well as up to six or more additional chains, such as CD3δ, CD3γ, CD3ε, and CD3ζ, although the exact composition of the T cell receptor complex can vary. The T cell receptor complex mediates intracellular signaling in T cells, which can lead to T cell activation.

[0050] The term "antibody" encompasses any antibody protein construct containing at least one antibody variable domain, which includes at least one antigen-binding site (ABS). Antibodies include, but are not limited to, IgA, IgG, IgE, IgD, and IgM type immunoglobulins (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).

[0051] Typical antibodies or immunoglobulins (Ig) are proteins containing 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 associated domains, and domains derived from them may be referred herein as immunoglobulin chain variable domains. The VH domains and VL domains (also referred to as VH regions and VL regions) may be further divided into regions called "complementarity-determining regions" ("CDRs"), which are interspersed with more conserved regions called "framework regions" ("FRs"). Framework regions and complementarity-determining regions are precisely defined (Kabat et al. Sequences of Proteins of Immunological Interest, Fifth Edition USD Department of Health and Human Services, (1991) NIH Publication Number 91-3242). Other numbering rules for CDR sequences exist, e.g., described in Chothia et al. (1989) Nature 342:877-883, or compiled by IMGT.org. In typical antibodies, each VH and VL consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. A typical antibody tetramer of two immunoglobulin heavy chains and two immunoglobulin light chains is formed, for example, by immunoglobulin heavy and light chains linked together by disulfide bonds, and similarly linked heavy chains. The constant region of the heavy chain contains three domains: CH1, CH2, and CH3. The constant region of the light chain consists of one domain: CL. The variable domains of the heavy chain and light chain are binding domains that interact with the antigen. The constant region of an antibody typically mediates 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.

[0052] As used herein, an antibody “fragment” (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 such a portion) that specifically binds to the δ variable region 1 (Vδ1) chain of a target γδ T cell receptor (for example, a molecule in which one or more immunoglobulin chains are not full length but specifically bind to the target). Examples of binding fragments encompassed by the term antibody fragment include: (i) Fab fragment (a monovalent fragment consisting of VL, VH, CL, and CH1 domains); (ii) F(ab′)2 fragment (a divalent fragment consisting of two Fab fragments linked by disulfide bridges in the hinge region); (iii) Fd fragment (consisting of VH and CH1 domains); (iv) Fv fragment (consisting of the VL and VH domains of a single arm of the antibody); (v) Single-stranded variable fragment (scFv) (consisting of VL and VH domains linked by a synthetic linker, which allows the VL and VH regions to pair up using a recombination method to be produced as a single protein chain forming a monovalent molecule); (vi) VH (variable domain of immunoglobulin chain consisting of VH domains); (vii) VL (variable domain of immunoglobulin chain consisting of VL domains); (viii) Domain antibody (dAb) (consisting of either the VH or VL domain); (ix) minibody (consisting of a pair of scFv fragments linked by CH3 domains); and (x) Diabody (consisting of a non-covalent dimer of scFv fragments, each consisting of a VH domain from one antibody linked to a VL domain from another antibody by a small peptide linker).

[0053] "Human antibody" refers to an antibody having a variable region and a constant region derived from a human germline immunoglobulin sequence. Human subjects administered with the above human antibody do not develop a heterologous antibody response to the primary amino acids contained in the antibody (e.g., referred to as the HAMA reaction (human anti-mouse antibody reaction)). The above human antibody may contain amino acid residues not encoded by the human germline immunoglobulin sequence (e.g., mutations introduced by random or site-directed mutagenesis, or somatic mutation) in the CDR, particularly CDR3. However, this term is not intended to encompass antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, has been transplanted into a human framework sequence. Human antibodies prepared, expressed, produced, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into host cells, antibodies isolated from a combination of recombinant human antibody libraries, antibodies isolated from transgenic animals (e.g., mice) with respect to human immunoglobulin genes, or antibodies prepared, expressed, produced, or isolated by any other means including splicing human immunoglobulin gene sequences with other DNA sequences, may also be referred to as “recombinant human antibodies.”

[0054] Substituting at least one amino acid residue in the framework region of a non-human immunoglobulin variable domain with a corresponding residue derived from a human variable domain is called "humanization." Humanization of the variable domain may reduce immunogenicity in humans.

[0055] "Specificity" refers to the number of different types of antigens or antigenic determinants to which a particular antibody or its antigen-binding fragment 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 well-understood term in the art. A molecule is said to exhibit "specific binding" if it reacts with a particular target antigen or epitope more frequently, more rapidly, for a longer duration, and / or with higher affinity than it would with other targets. An antibody "specifically binds" to a target antigen or epitope if it binds with higher affinity, higher binding activity, more easily, and / or for a longer duration than it would with other substances.

[0056] The antibodies of the present invention include monospecific antibodies (i.e., antibodies that bind to only one antigen) and multispecific antibodies. A "multispecific antibody" is an antibody that can bind to multiple different epitopes simultaneously or sequentially. Generally, these epitopes do not exist on the same antigen. Therefore, multispecific antibodies have the ability to selectively bind to epitopes present on different antigens through multiple different binding domains. This is in contrast to conventional monospecific antibodies that do not have this ability. More precisely, a "monospecific antibody" specifically binds to only one antigen, but may have multiple binding sites for this one antigen (for example, a fully human IgG antibody has a titer of 2, and other antibodies may have higher titers, but if the antibody recognizes only one antigen, it is still classified as a monospecific antibody). Therefore, the multispecific antibodies of the present invention bind to multiple different antigens simultaneously and / or sequentially.

[0057] In some embodiments of the present invention, the antibody is a bispecific antibody. A "bispecific antibody" is an antibody that can bind simultaneously and / or sequentially to two different epitopes. Generally, these epitopes are not present on the same antigen. Therefore, a bispecific antibody has the ability to selectively bind to two different epitopes present on two different antigens via two different binding domains. This is in contrast to conventional monospecific antibodies that do not have this ability. Thus, the bispecific antibody of the present invention binds simultaneously and / or sequentially to two different antigens.

[0058] "Affinity," expressed by the equilibrium constant (KD) for the dissociation of an antigen and an antigen-binding polypeptide, is a measure of the binding strength between the antigenic determinant and the antigen-binding site on the antibody (or its antigen-binding fragment). A smaller KD value indicates a stronger binding strength between the antigenic determinant and the antigen-binding polypeptide. Alternatively, affinity can also be expressed as the affinity constant (KA), which is 1 / KD. Affinity can be determined by known methods depending on the specific antigen of interest. For example, KD can be determined by surface plasmon resonance.

[0059] 10 -6 Any KD value less than 1 is considered to indicate binding. Specific binding of an antibody or its antigen-binding fragment to an antigen or antigenic determinant can be measured by any preferred known method, including, for example, scatchard analysis and / or competitive binding assays, such as radioimmunoassay (RIA), enzyme immunoassay (EIA), and sandwich competitive assays, equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance, or spectroscopy (e.g., using fluorescence assays), as well as various variations thereof known in the art.

[0060] "Binding activity" is a measure of the strength of binding between an antibody or its antigen-binding fragment and the relevant antigen. Binding activity is related to both the affinity between the antigenic determinant and its antigen-binding site on the antibody, and the number of relevant binding sites present on the antibody.

[0061] "In vivo primordial" means being in its natural or original location, rather than being moved to another location. For example, a patient's in vivo primordial Vδ1+ cells refer to in vivo Vδ1 cells, not in vitro or ex vivo cells.

[0062] "Human tissue Vδ1+ cells," as well as "hematopoietic and blood Vδ1+ cells" and "tumor-infiltrating lymphocytes (TILs) Vδ1+ cells," are defined as Vδ1+ cells contained in or derived from either human tissue, the hematopoietic blood system, or a human tumor, respectively. All of the above cell types can be identified by (i) their location or place of origin, and (ii) their Vδ1+ TCR expression.

[0063] A “regulating antibody” is an antibody that, upon contact with or binding to a cell expressing a target to which the antibody binds, induces a measurable change, including, but not limited to, a measurable change in the cell cycle, and / or cell number, and / or cell viability, and / or the secretion of one or more cell surface markers, and / or one or more secretory molecules (e.g., cytokines, chemokines, leukotrienes), and / or function (e.g., cytotoxicity against target cells or diseased cells). A method of “regulating” a cell or population of cells means a method by which at least one measurable change in a cell or a group of cells or their secretion is induced to produce one or more “regulated cells.”

[0064] "Immune response" is a measurable change in at least one cell, one cell type, one endocrine pathway, or one exocrine pathway of the immune system (including, but not limited to, cellular, humoral, cytokine, and chemokine responses) upon the addition of a regulatory antibody.

[0065] "Immune cell" is defined as a cell of the immune system including, but not limited to, CD34+ cells, B cells, CD45+ (common leukocyte antigen) cells, αβ T cells, cytotoxic T cells, helper T cells, plasma cells, neutrophils, monocytes, macrophages, erythrocytes, platelets, dendritic cells, phagocytes, granulocytes, innate lymphoid cells, natural killer (NK) cells, and γδ T cells. Typically, combinatorial cell surface molecule analysis (e.g., by flow cytometry) is used to classify immune cells, identify immune cells, or group or cluster them into subpopulations for distinction. These can then be further subdivided by additional analysis. For example, CD45+ lymphocytes can be further subdivided into a vδ-positive population and a vδ-negative population.

[0066] "Model system" is a biological model or biological representation designed to help understand how well a pharmaceutical agent such as an antibody or its antigen-binding fragment can function as a medicament in ameliorating the signs or symptoms of a disease. Such models typically involve the use of in vitro, ex vivo, and in vivo diseased cells, non-diseased cells, healthy cells, effector cells, tissues and the like, in which the performance of the above medicaments is studied and compared.

[0067] "Diseased cell" exhibits a phenotype associated with the progression of a disease such as cancer, an infection such as a viral infection, or an inflammatory condition or inflammatory disease. For example, the diseased cell can be a tumor cell, an autoimmune tissue, or a virus-infected cell. Accordingly, the above diseased cell can be defined as tumorous, virus-infected, or inflammatory.

[0068] "Healthy cell" refers to a normal cell that is not afflicted with a disease. These may also be referred to as "normal" cells or "non-diseased" cells. Non-diseased cells include non-cancerous cells, non-infected cells, or non-inflammatory cells. The above cells are often used together with the associated diseased cells to measure the diseased cell specificity conferred by the agent and / or to better understand the therapeutic index of the agent.

[0069] "Pathogenic cell specificity" is a measure of how effectively effector cells or populations of effector cells (e.g., Vδ1+ cell populations) are able to distinguish and kill diseased cells, such as cancer cells, while preserving non-pathogenic or healthy cells. This ability can be measured in model systems and may involve comparing the tendency of effector cells or effector cell populations to selectively kill or lyse diseased cells to their ability to kill or lyse non-pathogenic or healthy cells. The above-mentioned pathogenic cell specificity can provide information about the potential therapeutic index of a drug.

[0070] "Enhanced lesion cell specificity" refers to the phenotype of effector cells or populations, such as Vδ1+ cells, that are regulated to further enhance their ability to specifically kill lesion cells. This enhancement can be measured in various ways, including a multiplier change or a percentage increase in the specificity or selectivity of lesion cell killing.

[0071] "ADCC," or "antibody-dependent cell-mediated cytotoxicity," refers to an immune response to cells coated with antibodies bound to cell surface antigens. This is a cell-mediated process in which immune effector cells (e.g., NK cells) recognize the antibody bound to the cell, causing degranulation and lysis of the target cell. Typically, this is mediated by Fc-Fcγ interactions. The Fc region of the antibody bound to the cell recruits effector cells (e.g., NK cells) expressing the Fcγ receptor, leading to degranulation of the effector cells and death of the target cells.

[0072] "Fc-effective" refers to an antibody containing a functional Fc region (crystallizable region), i.e., an Fc region that has not been mutated or otherwise deactivated. Fc-effective antibodies exhibit undiminished Fc function. Fc-effective antibodies may contain human IGHC heavy chain sequences listed in IMGT that have not been modified, manipulated, or constructed to reduce binding to one or more Fcγ receptors. For example, this may be due to mutations in the IGHC hinge or the construction of an antibody containing a heavy chain constant domain that is a chimeric or hybrid of an IgG1 / IgG2A or IgG1 / IgG4 IGHC sequence.

[0073] Preferably, the antibody or its antigen-binding fragment (i.e., polypeptide) of the present invention is isolated. “Isolated” polypeptide is one that has been removed from its original environment. The term “isolated” may be used to refer to an antibody that substantially does not contain other antibodies having different antigen specificities (for example, an isolated antibody or fragment that specifically binds to Vδ1 substantially does not contain antibodies that specifically bind to antigens other than Vδ1). The term “isolated” may be used to refer to a preparation in which the isolated antibody, if formulated as an active ingredient in a pharmaceutical composition, is pure enough to be administered therapeutically, or has a purity of at least 70–80% (w / w), more preferably at least 80–90% (w / w), even more preferably 90–95%, most preferably at least 95%, 96%, 97%, 98%, 99%, or 100% (w / w).

[0074] Preferably, the polynucleotides used in the present invention are isolated. “Isolated” polynucleotides are those taken from their original environment. For example, a naturally occurring polynucleotide is isolated if it is separated from some or all of the materials that coexist in the natural system. A polynucleotide is considered isolated if, for example, it is cloned into a vector that is not part of its natural environment, or if it is contained within cDNA.

[0075] An antibody or its antigen-binding fragment may be a "functionally active variant," and examples of functionally active variants include naturally occurring allele variants, as well as mutants or any other non-natural variants. As is known in the art, an allele variant is an alternative form of a (poly)peptide characterized by having 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 functionally active variant may still function even if the framework containing the CDR is modified, the CDR itself is modified, the CDR is grafted onto an alternative framework, or an N-terminal or C-terminal extension is incorporated. Furthermore, the CDR-containing binding domain may be paired with a different partner chain, such as one shared with another antibody. The binding domain may still function even when shared with a so-called "common" light chain or "common" heavy chain. Furthermore, the binding domain may function when polymerized. Furthermore, “antibodies or their antigen-binding fragments” may also include functional variants that have been modified to move away from or approach a different canonical sequence (e.g., those listed on IMGT.org) with a different VH, VL, or constant domain, and that remain functional.

[0076] To compare two closely related polypeptide sequences, the "% sequence identity" between the first and second polypeptide sequences can be calculated using NCBI BLAST v2.0 with a standard setting on polypeptide sequences (BLASTP). To compare two closely related polynucleotide sequences, the "% sequence identity" between the first and second polynucleotide sequences can be calculated using NCBI BLAST v2.0 with a standard setting on nucleotide sequences (BLASTN).

[0077] Polypeptide or polynucleotide sequences are said to be the same as, or "identical to," another polypeptide or polynucleotide sequence if they share 100% sequence identity throughout their entire length. Residues in a sequence are numbered from left to right, i.e., from the N-terminus to the C-terminus in polypeptides; and from the 5' end to the 3' end in polynucleotides.

[0078] In some embodiments, any particular % sequence identity of the sequence is calculated without all six CDR sequences of the antibody. For example, an anti-Vδ1 antibody or its antigen-binding fragment may contain a heavy chain variable region sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with a particular heavy chain variable region sequence, and / or a light chain variable region sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with a particular light chain variable region sequence, where any amino acid diversity arises only in the heavy chain variable region sequence and the framework region of the light chain variable region sequence. In such embodiments, an anti-Vδ1 antibody or its antigen-binding fragment having a particular sequence identity retains the complete CDR1, CDR2, and CDR3 sequences of the heavy and light chains of the corresponding anti-Vδ1 antibody or its antigen-binding fragment. More specific examples are provided, not to limit, but solely to further illustrate these embodiments of the present invention, including an anti-Vδ1 antibody or its antigen-binding fragment comprising an amino acid sequence, or consisting thereof, that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15, and an amino acid sequence, or consisting thereof, that is at least 90% identical to the amino acid sequence of SEQ ID NO: 40, where any amino acid diversity arises only in the framework regions of the heavy chain variable region sequence and the light chain variable region sequence. Accordingly, the antibodies of this specific example further include VHCDR1 comprising the amino acid sequence of SEQ ID NO: 51, VHCDR2 comprising the amino acid sequence of SEQ ID NO: 53, VHCDR3 comprising the amino acid sequence of SEQ ID NO: 68, VLCDR1 comprising the amino acid sequence of SEQ ID NO: 79, VLCDR2 comprising the amino acid sequence of SEQ ID NO: 80, and VLCDR3 comprising the amino acid sequence of SEQ ID NO: 95.

[0079] Furthermore, the antibodies and antigen-binding fragments provided herein include a κ light chain variable sequence and may hold an amino acid residue at position 74, which is not serine, according to the IMGT numbering system, for example, a non-polar and / or non-germline type residue, for example, they include a leucine residue at this position. For example, and not to limit, but solely to further illustrate these embodiments of the present invention, an anti-Vδ1 antibody or antigen-binding fragment is provided, comprising VH, which includes an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 15, and VL, which includes an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 40, where any amino acid diversity arises only in the framework region of the heavy chain variable region sequence and the light chain variable region sequence, and the antibody includes a κ light chain variable sequence that includes an amino acid residue at position 74, which is non-germline type and / or non-polar according to the IMGT numbering system (for example, a leucine residue at this position). This specific example of an antibody further includes VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, VHCDR3 containing the amino acid sequence of SEQ ID NO: 68, VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 95.

[0080] 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 may contain one, two, or more such amino acid differences. An insertion, deletion, or substitution in the second sequence that is otherwise identical to the first sequence (100% sequence identity) results in a decrease in sequence identity percentage. For example, if the identical sequences are 9 amino acid residues long, one substitution in the second sequence results in 88.9% sequence identity. If the first and second polypeptide sequences are 9 amino acid residues long and share 6 identical residues, the first and second polypeptide sequences share more than 66% identity (they share 66.7% identity).

[0081] Alternatively, to compare 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 identified. “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). “Substitution” is the substitution of one amino acid residue in the sequence of the first polypeptide with one different amino acid residue. Such substitutions may be conserved or non-conservative. “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).

[0082] Using three-letter and one-letter codes, naturally occurring amino acids can be named as follows: glycine (G or Gly), alanine (A or Ala), valine (V or Val), leucine (L or Leu), isoleucine (I or Ile), proline (P or Pro), phenylalanine (F or Phe), tyrosine (Y or Tyr), tryptophan (W or Trp), lysine (K or Lys), arginine (R or Arg), histidine (H or His), aspartic acid (D or Asp), glutamic acid (E or Glu), asparagine (N or Asn), glutamine (Q or Gln), cysteine ​​(C or Cys), methionine (M or Met), serine (S or Ser), and threonine (T or Thr). If the residue may be aspartic acid or asparagine, the symbols Asx or B may be used. If the residue can be any amino acid, the symbols Xaa or X may be used. If the residue can be glutamic acid or glutamine, the symbols Glx or Z may be used. Unless otherwise specified in the context, a reference to aspartic acid includes aspartate, and a reference to glutamic acid includes glutamate.

[0083] As used herein, polypeptide sequence numbering and the definitions of CDR and FR are as defined in accordance with the EU and / or IMGT numbering systems, as indicated in the context. A “corresponding” amino acid residue between a first polypeptide sequence and a second polypeptide sequence is, as indicated in the context, an amino acid residue in the first sequence that shares the same position as an amino acid residue in the second sequence, according to the EU and / or IMGT numbering system, although such amino acid residue in the second sequence may differ in identity from that in the first sequence. Preferably, corresponding residues share the same number (and letter) if the framework and CDR are of the same length according to the EU or IMGT definition. Alignment can be achieved manually or by using known computer algorithms for sequence alignment, such as NCBI BLAST v2.0 (BLASTP or BLASTN) using standard settings.

[0084] In this specification, the term “epitope” refers to a portion of a target to which an antibody or its antigen-binding fragment specifically binds. An epitope may also be called an “antigenic determinant.” An antibody binds to “essentially the same epitope” as another antibody if both recognize the same or sterically overlapping epitopes. A method commonly used to determine whether two antibodies bind to the same or overlapping epitopes is a competitive assay, which can consist of a number of different formats using either a labeled antigen or a labeled antibody (e.g., well plates using radiolabeling or enzymatic labeling, or flow cytometry in cells expressing the antigen). An antibody binds to “the same epitope” as another antibody if both recognize the same epitope (i.e., all contact points between the antigen and the antibody are the same).

[0085] Epitopes found in protein targets can be defined as "linear epitopes" or "structural epitopes." Linear epitopes are formed by a continuous sequence of amino acids in a protein antigen. Structural epitopes are formed from amino acids that are discontinuous within the protein sequence but assemble when the protein folds into a three-dimensional structure.

[0086] As used herein, the term “vector” is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it is ligated. One type of vector is a “plasmid,” which refers to a circular double-stranded DNA loop to which an additional DNA fragment can be ligated. Another type of vector is a viral vector to which an additional DNA fragment can be ligated into a viral genome. Certain vectors are capable of self-replication within the host cell into which they are introduced (e.g., bacterial vectors with bacterial replication origins, episomal mammalian vectors, and yeast vectors). Other vectors (e.g., non-episomal mammalian vectors) can be incorporated into the host cell's genome upon introduction into the host cell, thereby replicating with the host genome. Furthermore, certain types of vectors can induce the expression of genes to which they are operably ligated. Such vectors are referred herein as “recombinant expression vectors,” or simply “expression vectors.” Generally, expression vectors useful in recombinant DNA technology are often in the form of plasmids. Since plasmids are the most commonly used form of vector, “plasmid” and “vector” may be used interchangeably herein. However, the present invention is intended to encompass viral vectors that perform equivalent functions (e.g., replication-deficient retroviruses, adenoviruses, and adeno-associated viruses), as well as other forms of expression vectors such as bacteriophages and phagemid systems. As used herein, the term “recombinant host cell” (or simply “host cell”) is intended to refer to a cell into which a recombinant expression vector has been introduced. Such term is intended to refer not only to a specific target cell, but also, for example, if the offspring of such cells are used to create a cell line or cell bank, and these are then optionally stored, provided, sold, transferred, or used to produce antibodies or antigen-binding fragments described herein.

[0087] References to “subject,” “patient,” or “individual” refer to the subject being treated, particularly mammalian subjects. Mammalian subjects include humans, non-human primates, livestock (e.g., cattle), sports animals, or companion animals, such as dogs, cats, guinea pigs, rabbits, rats, or mice. In some embodiments, the subject is human. In alternative embodiments, the subject is a non-human mammal such as a mouse.

[0088] The term "sufficient dose" means an amount sufficient to produce the desired effect. The term "therapeutic dose" is an amount that is effective in improving the symptoms of a disease or disorder. A therapeutic dose may be a "prophylactic dose" if prevention can be considered a treatment.

[0089] A disease or disorder is considered "mitigated" if the severity of its signs or symptoms, the frequency with which the subject experiences such signs or symptoms, or both, is reduced (compared to an initial point in time, e.g., before administration of any antibody).

[0090] As used herein, “to treat a disease or disorder” means to reduce the frequency and / or severity of at least one sign or symptom of a disease or disorder experienced by the subject (compared to an earlier point in time, e.g., before administration of any antibody).

[0091] As used herein, “cancer” refers to the abnormal proliferation or division of cells. Generally, the proliferation and / or lifespan of cancer cells outpaces and does not coordinate with the proliferation and / or lifespan of the surrounding normal cells and tissues. Cancer can be benign, premalignant, or malignant. Cancer can occur in various cells and tissues, including those of the oral cavity (e.g., mouth, tongue, pharynx), digestive system (e.g., esophagus, stomach, small intestine, colon, rectum, liver, bile duct, gallbladder, pancreas), respiratory system (e.g., larynx, lungs, bronchi), bones, joints, skin (e.g., basal cells, squamous cells, meningiomas), breasts, reproductive system (e.g., uterus, ovaries, prostate, testes), urinary system (e.g., bladder, kidneys, ureters), eyes, nervous system (e.g., brain), endocrine system (e.g., thyroid), and hematopoietic system (e.g., lymphoma, myeloma, leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia).

[0092] As used herein, the term “about” includes values ​​up to 10% higher (inclusive) and 10% lower (inclusive) than a specified value, preferably including values ​​up to 5% higher (inclusive) and 5% lower (inclusive) than a specified value, and specifically including the specified value. The term “between” includes the specified boundary values.

[0093] Antibodies and their antigen-binding fragments This specification provides an antibody or an antigen-binding fragment thereof that can specifically bind to the δ variable region 1 (Vδ1) of the γδ T cell receptor (TCR). The present invention relates to the use of the above antibody as a drug for administration to a subject to be treated.

[0094] In one embodiment, the antibody or its antigen-binding fragment is an scFv, Fab, Fab', F(ab')2, Fv, a variable domain (e.g., VH or VL), a diabody, a minibody, or a monoclonal antibody. In a further embodiment, the antibody or its antigen-binding fragment is an scFv.

[0095] The antibodies of the present invention may be antibodies of any class, e.g., IgG, IgA, IgM, IgE, IgD, or their isotypes, and may contain a κ light chain or a λ 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 further embodiments, the antibody may be in a format such as an IgG format, having a mutated Fc that has been modified to confer a desired property, e.g., to reduce effector function, to extend half-life, to alter ADCC, or to improve hinge stability. Such modifications are well known in the art.

[0096] In one embodiment, the antibody or its antigen-binding fragment is human. Therefore, the antibody or its antigen-binding fragment may be derived from a human immunoglobulin (Ig) sequence. The CDR, framework, and / or constant region of the antibody (or its antigen-binding fragment) may be derived from a human Ig sequence, particularly a human IgG sequence. The CDR, framework, and / or constant region may be substantially identical with respect to a human Ig sequence, particularly a human IgG sequence. The advantage of using human antibodies is that they are low-immunogenic or non-immunogenic in humans.

[0097] The antibody or its antigen-binding fragment may be a chimeric antibody, for example, a mouse / human chimeric antibody.

[0098] Alternatively, the antibody or its antigen-binding fragment may be derived from a non-human species, such as a mouse. Such a non-human antibody may be modified to increase its similarity to antibody variants naturally produced in humans, thereby partially or completely humanizing the antibody or its antigen-binding fragment. Thus, in one embodiment, the antibody or its antigen-binding fragment is humanized.

[0099] Overview of the specific antibodies provided herein A partial overview of some of the specific antigen-binding molecules (i.e., antibodies) provided by the present invention is shown below, along with their assigned sequence numbers in the accompanying sequence listing. Antigen-binding variants, derivatives, and fragments thereof are also provided as part of the present invention. The sequences are shown in the accompanying sequence listing and accompanying figures. In the event of any discrepancy between the sequences in the sequence listing and the sequences in Figures 45-48, the sequences in the figures shall prevail. [Table 1-1] [Table 1-2] [Table 2-1] [Table 2-2] [Table 3] [Table 4] [Table 5]

[0100] Antibodies derived from ADT1-4 and ADT1-7 The present invention provides antibodies derived from the parent antibody ADT1-4 (having the heavy chain variable region sequence described in SEQ ID NO: 1 and the light chain variable region sequence described in SEQ ID NO: 26), and antibodies derived from the parent antibody ADT1-7 (having the heavy chain variable region sequence described in SEQ ID NO: 106 and the light chain variable region sequence described in SEQ ID NO: 118). ADT1-4 is also referred to herein as G04, and ADT1-4 and G04 are used interchangeably. ADT1-7 is also referred to herein as E07, and ADT1-7 and E07 are used interchangeably.

[0101] In some embodiments, the present invention provides an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising: A heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NOs. 55-78 and 133-143; and / or a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NOs. 82-105 and 147-157. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0102] In some embodiments, the present invention provides an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising: VHCDR1 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs. 51, 52, and 130; VHCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs. 53 and 131; and A heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NOs. 55-78 and 133-143, and VHCDR3 containing a heavy chain variable region comprising such an amino acid sequence, and VLCDR1 comprising, or consisting thereof, an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs. 79 and 144; VLCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs. 80 and 145; and A light chain variable region containing VLCDR3 that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NOs.82-105 and 147-157.

[0103] Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0104] In some embodiments, the present invention provides an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising: A heavy chain variable region containing or consisting of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence selected from the group consisting of SEQ ID NOs. 2-25 and 107-117; and / or A light chain variable region containing or consisting of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with a sequence selected from the group consisting of sequence numbers 27-50 and 119-129.

[0105] Antibodies derived from ADT1-4 The present invention provides an antibody derived from the parent antibody ADT1-4 (having the heavy chain variable region sequence described in SEQ ID NO: 1 and the light chain variable region sequence described in SEQ ID NO: 26), as described below, for example. ADT1-4 is also referred to as G04 herein, and ADT1-4 and G04 are used interchangeably.

[0106] Antibodies containing specific CDR sequences derived from ADT1-4 The antibodies provided herein include the following antibodies having specific sequences derived from ADT1-4.

[0107] For example, in some embodiments, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 55 to 78, or a VHCDR3 comprising such sequence; and / or a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 82 to 105, or a VLCDR3 comprising such sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0108] In some embodiments, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment is provided, comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 55 to 77, or a VHCDR3 comprising such sequence; and / or a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 82 to 104, or a VLCDR3 comprising such sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0109] In some embodiments, an anti-TCRδ variable region 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof is provided, comprising a heavy chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 58, 60, 61, 62, 65, 66, 68, 74, 76, and 77, or a VHCDR3 comprising such sequence; and / or a light chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 85, 87, 88, 89, 92, 93, 95, 101, 103, and 104, or a VLCDR3 comprising such sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0110] Anti-TCRδ variable region 1 (anti-Vδ1) antibodies or their antigen-binding fragments, including the following, are also provided: VHCDR1 containing or consisting of an amino acid sequence selected from the group comprising SEQ ID NOs. 51 and 52; VHCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 53; and A heavy chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 55-78, or a VHCDR3 comprising such a sequence, and VLCDR1 containing or consisting of the amino acid sequence of SEQ ID NO: 79; VLCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 80; and A light chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 82-105, or a VLCDR3 comprising such a sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0111] Anti-TCRδ variable region 1 (anti-Vδ1) antibodies or their antigen-binding fragments, including the following, are also provided: VHCDR1 containing or consisting of an amino acid sequence selected from the group comprising SEQ ID NOs. 51 and 52; VHCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 53; and A heavy chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 55 to 77, or a VHCDR3 comprising such a sequence, and VLCDR1 containing or consisting of the amino acid sequence of SEQ ID NO: 79; VLCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 80; and A light chain variable region containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 82-104, or a VLCDR3 comprising such a sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0112] Anti-TCRδ variable region 1 (anti-Vδ1) antibodies or their antigen-binding fragments, including the following, are also provided: VHCDR1 containing or consisting of an amino acid sequence selected from the group comprising SEQ ID NOs. 51 and 52; VHCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 53; and A heavy chain variable region containing VHCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 58, 60, 61, 62, 65, 66, 68, 74, 76, and 77, and VLCDR1 containing or consisting of the amino acid sequence of SEQ ID NO: 79; VLCDR2 containing or consisting of the amino acid sequence of SEQ ID NO: 80; and A light chain variable region containing a VLCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs. 85, 87, 88, 89, 92, 93, 95, 101, 103, and 104.

[0113] Certain amino acid substitutions may be made to provide one or more variant antibodies described herein.

[0114] Anti-TCRδ variable region 1 (anti-Vδ1) antibodies or their antigen-binding fragments, including the following, are also provided: a) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 55, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 82, respectively; b) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 56, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 83, respectively; c) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 57, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 84, respectively; d) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 58, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 85, respectively; e) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 59, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 86, respectively; f) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 60, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 87, respectively; g) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 52, 53, and 61, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 88, respectively; h) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 62, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 89, respectively; i) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 63, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 90, respectively; j) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 64, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 91, respectively; k) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 65, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 92, respectively; l) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 52, 53, and 66, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 93, respectively; m) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 67, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 94, respectively; n) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 68, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 95, respectively; o) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 69, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 96, respectively; p) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 70, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 97, respectively; q) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 71, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 98, respectively; r) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 72, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 99, respectively; s) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 73, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 100, respectively; t) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 52, 53, and 74, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 101, respectively; u) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 75, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 102, respectively; v) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 76, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 103, respectively; w) VHCDR1, VHCDR2, and VHCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 51, 53, and 77, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing or consisting of the amino acid sequences of SEQ ID NOs. 79, 80, and 104, respectively; or x) VHCDR1, VHCDR2, and VHCDR3 each containing or comprising the amino acid sequences of SEQ ID NOs. 51, 53, and 78, respectively, and VLCDR1, VLCDR2, and VLCDR3 each containing or comprising the amino acid sequences of SEQ ID NOs. 79, 80, and 105, respectively.

[0115] The anti-Vδ1 antibody or its antigen-binding fragment may contain a κ light chain variable sequence (or a variable light chain derived from a κ light chain variable sequence), where, according to the IMGT numbering system, the residue at position 74 of the κ light chain variable sequence is not serine, but rather, for example, a non-human germline residue and / or nonpolar residue at position 74, for example, a leucine residue.

[0116] Further embodiments are provided below.

[0117] ADT1-4-105 and its fragments and variants Certain embodiments relate to the antibody ADT1-4-105 and its fragments and variants.

[0118] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 55, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 82. In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising a heavy chain variable region containing VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 55, and / or a light chain variable region containing VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 82.

[0119] To provide a variant antibody derived from ADT1-4-105, amino acid substitutions may be made, for example, an antibody or antigen-binding fragment or variant thereof is provided, comprising a heavy chain variable region containing VHCDR3 with the amino acid sequence of SEQ ID NO: 55, optionally containing one or two amino acid substitutions, and / or a light chain variable region containing VLCDR3 with the amino acid sequence of SEQ ID NO: 82, optionally containing one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0120] In some embodiments, an antibody or its antigen-binding fragment or variant is provided, comprising the following: VHCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 51, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, or less identity to the amino acid sequence of SEQ ID NO: 53 A heavy chain variable region including VHCDR2 containing at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55, and VHCDR3 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and / or VLCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 80, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% identity to the amino acid sequence of SEQ ID NO: 80 A light chain variable region comprising VLCDR2 having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 82, and VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 82.

[0121] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 51, VHCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 53, and VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 55; and / or A light chain variable region comprising VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 79, VLCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 80, and VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 82.

[0122] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, and VHCDR3 containing the amino acid sequence of SEQ ID NO: 55; and A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 82.

[0123] Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-105, and for example, antibodies or antigen-binding fragments or variants thereof are provided, including the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, which optionally contains one or two amino acid substitutions; VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, which optionally contains one or two amino acid substitutions; and VHCDR3 containing the amino acid sequence of SEQ ID NO: 55, which optionally contains one or two amino acid substitutions; and / or A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, which optionally contains one or two amino acid substitutions; VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, which optionally contains one or two amino acid substitutions; and VLCDR3 containing the amino acid sequence of SEQ ID NO: 82, which optionally contains one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0124] Alternatively, the antibody may consist of a specified sequence (with or without amino acid substitutions).

[0125] ADT1-4-107 and its fragments and variants Specific embodiments relate to the antibody ADT1-4-107 and its fragments and variants.

[0126] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 56, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 83. In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising a heavy chain variable region containing VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 56, and / or a light chain variable region containing VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 83.

[0127] To provide a variant antibody derived from ADT1-4-107, amino acid substitutions may be made, for example, an antibody or antigen-binding fragment or variant thereof is provided, comprising a heavy chain variable region containing VHCDR3 with the amino acid sequence of SEQ ID NO: 56, optionally containing one or two amino acid substitutions, and / or a light chain variable region containing VLCDR3 with the amino acid sequence of SEQ ID NO: 83, optionally containing one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0128] In some embodiments, an antibody or its antigen-binding fragment or variant is provided, comprising the following: VHCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 51, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, or less identity to the amino acid sequence of SEQ ID NO: 53 A heavy chain variable region including VHCDR2 containing at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56, and VHCDR3 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56; and / or VLCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 80, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% identity to the amino acid sequence of SEQ ID NO: 80 A light chain variable region comprising VLCDR2 having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 83, and VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 83.

[0129] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 51, VHCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 53, and VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 56; and / or A light chain variable region comprising VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 79, VLCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 80, and VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 83.

[0130] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: Heavy chain variable regions including VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, and VHCDR3 containing the amino acid sequence of SEQ ID NO: 56; and A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 83.

[0131] Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-107, and for example, antibodies or antigen-binding fragments or variants thereof are provided, including the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, which optionally contains one or two amino acid substitutions; VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, which optionally contains one or two amino acid substitutions; and VHCDR3 containing the amino acid sequence of SEQ ID NO: 56, which optionally contains one or two amino acid substitutions; and / or A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, which optionally contains one or two amino acid substitutions; VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, which optionally contains one or two amino acid substitutions; and VLCDR3 containing the amino acid sequence of SEQ ID NO: 83, which optionally contains one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0132] Alternatively, the antibody may consist of a specified sequence (with or without amino acid substitutions).

[0133] ADT1-4-110 and its fragments and variants Specific embodiments relate to the antibody ADT1-4-110 and its fragments and variants.

[0134] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 57, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 84. In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising a heavy chain variable region containing VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 57, and / or a light chain variable region containing VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 84.

[0135] To provide a variant antibody derived from ADT1-4-110, amino acid substitutions may be made, for example, an antibody or antigen-binding fragment or variant thereof is provided, comprising a heavy chain variable region containing VHCDR3 with the amino acid sequence of SEQ ID NO: 57, optionally containing one or two amino acid substitutions, and / or a light chain variable region containing VLCDR3 with the amino acid sequence of SEQ ID NO: 84, optionally containing one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0136] In some embodiments, an antibody or its antigen-binding fragment or variant is provided, comprising the following: VHCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 51, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, or less identity to the amino acid sequence of SEQ ID NO: 53 A heavy chain variable region including VHCDR2 containing at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 57, and VHCDR3 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 57; and / or VLCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 80, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% identity to the amino acid sequence of SEQ ID NO: 80 A light chain variable region comprising VLCDR2 having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 84, and VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 84.

[0137] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 51, VHCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 53, and VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 57; and / or A light chain variable region comprising VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 79, VLCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 80, and VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 84.

[0138] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: Heavy chain variable regions including VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, and VHCDR3 containing the amino acid sequence of SEQ ID NO: 57; and A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 84.

[0139] Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-110, and for example, antibodies or antigen-binding fragments or variants thereof are provided, including the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, which optionally contains one or two amino acid substitutions; VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, which optionally contains one or two amino acid substitutions; and VHCDR3 containing the amino acid sequence of SEQ ID NO: 57, which optionally contains one or two amino acid substitutions; and / or A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, which optionally contains one or two amino acid substitutions; VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, which optionally contains one or two amino acid substitutions; and VLCDR3 containing the amino acid sequence of SEQ ID NO: 84, which optionally contains one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0140] Alternatively, the antibody may consist of a specified sequence (with or without amino acid substitutions).

[0141] ADT1-4-112 and its fragments and variants Specific embodiments relate to the antibody ADT1-4-112 and its fragments and variants.

[0142] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 58, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 85. In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising a heavy chain variable region containing VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 58, and / or a light chain variable region containing VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 85.

[0143] To provide a variant antibody derived from ADT1-4-112, amino acid substitutions may be made, for example, an antibody or antigen-binding fragment or variant thereof is provided, comprising a heavy chain variable region containing VHCDR3 with the amino acid sequence of SEQ ID NO: 58, optionally containing one or two amino acid substitutions, and / or a light chain variable region containing VLCDR3 with the amino acid sequence of SEQ ID NO: 85, optionally containing one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0144] In some embodiments, an antibody or its antigen-binding fragment or variant is provided, comprising the following: VHCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 51, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, or less identity to the amino acid sequence of SEQ ID NO: 53 A heavy chain variable region including VHCDR2 containing at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 58, and VHCDR3 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 58; and / or VLCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 80, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% identity to the amino acid sequence of SEQ ID NO: 80 A light chain variable region comprising VLCDR2 having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 85, and VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 85.

[0145] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 51, VHCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 53, and VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 58; and / or A light chain variable region comprising VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 79, VLCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 80, and VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 85.

[0146] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: Heavy chain variable regions including VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, and VHCDR3 containing the amino acid sequence of SEQ ID NO: 58; and A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 85.

[0147] Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-112, and for example, antibodies or antigen-binding fragments or variants thereof are provided, including the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, which optionally contains one or two amino acid substitutions; VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, which optionally contains one or two amino acid substitutions; and VHCDR3 containing the amino acid sequence of SEQ ID NO: 58, which optionally contains one or two amino acid substitutions; and / or A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, which optionally contains one or two amino acid substitutions; VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, which optionally contains one or two amino acid substitutions; and VLCDR3 containing the amino acid sequence of SEQ ID NO: 85, which optionally contains one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0148] Alternatively, the antibody may consist of a specified sequence (with or without amino acid substitutions).

[0149] ADT1-4-117 and its fragments and variants Specific embodiments relate to the antibody ADT1-4-117 and its fragments and variants.

[0150] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 59, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 86. In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising a heavy chain variable region containing VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 59, and / or a light chain variable region containing VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 86.

[0151] To provide a variant antibody derived from ADT1-4-117, amino acid substitutions may be made, for example, an antibody or antigen-binding fragment or variant thereof is provided, comprising a heavy chain variable region containing VHCDR3 with the amino acid sequence of SEQ ID NO: 59, optionally containing one or two amino acid substitutions, and / or a light chain variable region containing VLCDR3 with the amino acid sequence of SEQ ID NO: 86, optionally containing one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0152] In some embodiments, an antibody or its antigen-binding fragment or variant is provided, comprising the following: VHCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 51, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, or less identity to the amino acid sequence of SEQ ID NO: 53 A heavy chain variable region including VHCDR2 containing at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 59, and VHCDR3 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 59; and / or VLCDR1 containing at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 80, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93% identity to the amino acid sequence of SEQ ID NO: 80 A light chain variable region comprising VLCDR2 having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 86, and VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 86.

[0153] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: A heavy chain variable region comprising VHCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 51, VHCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 53, and VHCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 59; and / or A light chain variable region comprising VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO: 79, VLCDR2 having at least 90% identity to the amino acid sequence of SEQ ID NO: 80, and VLCDR3 having at least 90% identity to the amino acid sequence of SEQ ID NO: 86.

[0154] In one embodiment, an antibody or its antigen-binding fragment or variant is provided, comprising the following: Heavy chain variable regions including VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, and VHCDR3 containing the amino acid sequence of SEQ ID NO: 59; and A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, and VLCDR3 containing the amino acid sequence of SEQ ID NO: 86.

[0155] Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-117, and for example, antibodies or antigen-binding fragments or variants thereof are provided, including the following: A heavy chain variable region comprising VHCDR1 containing the amino acid sequence of SEQ ID NO: 51, which optionally contains one or two amino acid substitutions; VHCDR2 containing the amino acid sequence of SEQ ID NO: 53, which optionally contains one or two amino acid substitutions; and VHCDR3 containing the amino acid sequence of SEQ ID NO: 59, which optionally contains one or two amino acid substitutions; and / or A light chain variable region comprising VLCDR1 containing the amino acid sequence of SEQ ID NO: 79, which optionally contains one or two amino acid substitutions; VLCDR2 containing the amino acid sequence of SEQ ID NO: 80, which optionally contains one or two amino acid substitutions; and VLCDR3 containing the amino acid sequence of SEQ ID NO: 86, which optionally contains one or two amino acid substitutions. The amino acid substitutions may be conservative amino acid substitutions.

[0156] Alternatively, the antibody may consist of a specified sequence (with or without amino acid substitutions).

[0157] ADT1-4-19 and its fragments and variants Specific embodiments relate to the antibody ADT1-4-19 and its fragments and variants.

[0158] For example, in some embodiments, an antibody or antigen-binding fragment or variant is provided that includes a heavy chain variable region containing VHCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 60, and / or an antibody or antigen-binding fragment or variant that includes a light chain variable region containing VLCDR3 having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 87. In one emb...

Claims

1. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, wherein the antibody or its antigen-binding fragment is a. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 68, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 95, respectively; b. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 55, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 82, respectively; c. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 56, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 83, respectively; d. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 57, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 84, respectively; e. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 58, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 85, respectively; f. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 59, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 86, respectively; g. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 60, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 87, respectively; h. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 61, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 88, respectively; i. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 62, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 89, respectively; j. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 63, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 90, respectively; k. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 64, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 91, respectively; l. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 65, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 92, respectively; m. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 66, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 93, respectively; n. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 67, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 94, respectively; o. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 69, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 96, respectively; p. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 70, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 97, respectively; q. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 71, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 98, respectively; r. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 72, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 99, respectively; s. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 73, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 100, respectively; t. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 74, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 101, respectively; u. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 75, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 102, respectively; v. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 76, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 103, respectively; w. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 77, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 104, respectively; or x. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 78, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 105, respectively. An antibody or its antigen-binding fragment.

2. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof according to claim 1, comprising a heavy chain variable region and a light chain variable region, wherein the antibody or the antigen-binding fragment thereof a. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 68, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 95, respectively; b. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 58, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 85, respectively; c. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 60, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 87, respectively; d. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 61, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 88, respectively; e. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 62, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 89, respectively; f. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 65, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 92, respectively; g. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 66, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 93, respectively; h. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 52, 53, and 74, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 101, respectively; i. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 76, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 103, respectively; j. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 77, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 104, respectively; or k. VHCDR1, VHCDR2, and VHCDR3 containing the amino acid sequences of SEQ ID NOs. 51, 53, and 78, respectively, and VLCDR1, VLCDR2, and VLCDR3 containing the amino acid sequences of SEQ ID NOs. 79, 80, and 105, respectively. An antibody or its antigen-binding fragment.

3. a. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 15, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40; b. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 2, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 27; c. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 3, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 28; d. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 4, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29; e. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 5, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 30; f. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 6, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31; g. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 7, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 32; h. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 8, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 33; i. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 9, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 34; j. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 10, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 35; k. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 11, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 36; l. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 12, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 37; m. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 13, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 38; n. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 14, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 39; o. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 16, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 41; p. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 17, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 42; q. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 18, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 43; r. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 19, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 44; s. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 20, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 45; t. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 21, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 46; u. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 22, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 47; v. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 23, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 48; w. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 24, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 49; or x. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 25, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:

50. The anti-Vδ1 antibody or its antigen-binding fragment according to claim 1 or 2.

4. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof according to claim 3, comprising a heavy chain variable region and a light chain variable region, wherein the antibody or the antigen-binding fragment thereof a. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 15, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40; b. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 5, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 30; c. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 7, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 32; d. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 8, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 33; e. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 9, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 34; f. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 12, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 37; g. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 13, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 38; h. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 21, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 46; i. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 23, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 48; j. VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 24, and VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 49; or k. A VH containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 25, and a VL containing an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:

50. An antibody or its antigen-binding fragment.

5. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising VH containing the amino acid sequence of SEQ ID NO: 15 and VL containing the amino acid sequence of SEQ ID NO:

40.

6. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising VH containing the amino acid sequence of SEQ ID NO: 23 and VL containing the amino acid sequence of SEQ ID NO:

48.

7. An anti-TCRδ variable region 1 (anti-Vδ1) antibody or its antigen-binding fragment, comprising VH containing the amino acid sequence of SEQ ID NO: 5 and VL containing the amino acid sequence of SEQ ID NO:

30.

8. The anti-Vδ1 antibody or antigen-binding fragment according to any one of claims 1 to 7, wherein the antibody or antigen-binding fragment comprises an amino acid residue other than serine at position 74 according to the IMGT numbering system of the κ light chain variable sequence, and optionally the residue at position 74 according to the IMGT numbering system is a leucine residue.

9. An anti-Vδ1 antibody or its antigen-binding fragment, wherein the antibody is a. The first target epitope, which is an epitope of the variable δ1 (Vδ1) chain of the γδ T cell receptor (TCR), and b. A multispecific antibody that specifically binds to a second target epitope, The multispecific antibody is distributed as follows: SEQ ID NO: 393; SEQ ID NO: 388; SEQ ID NO: 315; SEQ ID NO: 379; SEQ ID NO: 383; SEQ ID NO: 396; SEQ ID NO: 398; SEQ ID NO: 399; SEQ ID NO: 401; SEQ ID NO: 403; SEQ ID NO: 405; SEQ ID NO: 407; SEQ ID NO: 409; SEQ ID NO: 411; SEQ ID NO: 414 and SEQ ID NO: 415; SEQ ID NO: 414 and SEQ ID NO: 416; SEQ ID NO: 504 and SEQ ID NO: 419; SEQ ID NO: 504 and SEQ ID NO: 418; SEQ ID NO: 505 and SEQ ID NO: 415; SEQ ID NO: 505 and SEQ ID NO: 416; SEQ ID NO: 505 and SEQ ID NO: 419; SEQ ID NO: 505 and SEQ ID NO: 418; SEQ ID NO: 421 and SEQ ID NO: 422; The anti-Vδ1 antibody or antigen-binding fragment comprising sequence number 504 and sequence number 422; sequence number 421 and sequence number 437; sequence number 504 and sequence number 437; sequence number 421 and sequence number 429; sequence number 504 and sequence number 429; sequence number 414 and sequence number 431; sequence number 414 and sequence number 433; sequence number 414 and sequence number 434; sequence number 414 and sequence number 435; sequence number 504 and sequence number 431; sequence number 504 and sequence number 433; sequence number 504 and sequence number 434; sequence number 504 and sequence number 435; sequence number 505 and sequence number 431; sequence number 505 and sequence number 433; sequence number 505 and sequence number 434; sequence number 505 and sequence number 435; sequence number 438 and sequence number 417; or sequence number 439 and sequence number 420.

10. a. The antibody or its antigen-binding fragment causes depletion of less than 30%, less than 20%, or less than 10% of the viable Vδ1+ T cell population by ADCC and / or CDC; and / or b. The antibody or its antigen-binding fragment binds to human TRDV1 with a KD of less than approximately 10 nM, or optionally, the antibody or its antigen-binding fragment binds to cynomolgus monkey TRDV1 with a KD of less than approximately 100 nM. The anti-TCRδ variable portion 1 (anti-Vδ1) antibody or its antigen-binding fragment according to any one of claims 1 to 9.

11. The antibody or its antigen-binding fragment a. Bond affinity of less than approximately 100 nM (preferably less than approximately 50 nM) (for example, when measured by surface plasmon resonance, K Ｄ ) binds to human TRDV1 (sequence number 272 or 306), b. Optionally, a binding affinity of less than about 100 nM (preferably less than about 50 nM) (for example, when measured by surface plasmon resonance, K Ｄ ) binds to cynomolgus monkey TRDV1 (SEQ ID NO: 308), c. Having an IC50 that lowers the TCR to less than about 50 nM (preferably less than about 10 nM), d. Having an IC50 of less than approximately 10 nM (preferably less than approximately 5 nM) for THP-1 cell toxicity, e. An anti-Vδ1 antibody or antigen-binding fragment according to any one of claims 1 to 10, having an IC90 of less than approximately 50 nM for THP-1 cell toxicity.

12. The antibody or its antigen-binding fragment binds to an epitope in the amino acid region 37-77 of SEQ ID NO: 272, SEQ ID NO: 306, and / or SEQ ID NO:

308. An anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 11.

13. The anti-Vδ1 antibody or antigen-binding fragment according to claim 12, wherein the epitope is an activating epitope for γδ T cells, and optionally, binding of the activating epitope (i) downregulates the γδ TCR, (ii) activates degranulation of the γδ T cells, and / or (iii) promotes killing by γδ T cells; and / or upregulates the expression of CD107a, CD25, CD69, and / or Ki67.

14. The antibody is a multispecific antibody that specifically binds to the variable δ1 (Vδ1) chain of the γδ T cell receptor (TCR) and a second antigen, and the second antigen is a cancer antigen or cancer-related antigen, an immunomodulatory antigen, or a group of differentiation antigens (CD), optionally The aforementioned cancer antigen or cancer-related antigen is AFP, AKAP-4, ALK, α-fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCAA, Bcr-abl, β-catenin, β-hCG, β-human chorionic gonadotropin, BORIS, BTAA, CA125, CA15-3, CA195, CA19-9, CA242, CA27,29, CA72-4, 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, EL F2M, 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-related 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, Regmine, LMP2, M344, MA-50, Mac-2 Binding Protein, MAD-CT-1, MAD-CT-2, 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, Neuronal / 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 variant, Page4, PAX3, PAX5, PDGFR-β, PLAC1, Policy Auric acid, prostate cancer tumor antigen-1 (PCTA-1), prostate-specific antigen, prostatic acid phosphatase (PAP), proteinase 3 (PR1), PSA, PSCA, PSMA, RAGE-1, Ras, Ras variant, RCAS1, RGS5, RhoC, ROR1, RU1, RU2 (AS), SART3, SDCCAG16, sLe(a), sperm protein 17, SSX2, STn, Survivin, TA-90, TAAL6, TAG-72, telomerase, thyroglobulin, Tie 2. Selected from the group consisting of TLP, Tn, TPS, TRP-1, TRP-2, TRP-2, TSP-180, tyrosinase, VEGF, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72, The immunomodulatory antigen may be B7-1 (CD80), B7-2 (CD86), B7-DC (CD273), B7-H1 (CD274), B7-H2 (CD275), B7-H3 (CD276), B7-H4 (VTCN). 1), B7-H5 (VISTA), BTLA (CD272), CD137, CD137L, CD24, CD27, CD28, CD38, CD40, CD40L (CD154), CD54, CD59, CD70, C Selected from the group consisting of TLA4 (CD152), CXCL9, GITR (CD357), HVEM (CD270), ICAM-1 (CD54), ICOS (CD278), LAG-3 (CD223), OX40 (CD134), OX40L (CD252), PD-1 (CD279), PD-L1 (CD274), TIGIT, CD314, CD334, CD335, CD337, and TIM-3 (CD366), or The CD antigen is 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, 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, CD95, 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、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, CD275, CD276, CD277, C D278, CD279, CD280, CD281, CD282, CD283, CD284, CD285, CD286, CD287, CD288, CD289, CD290, CD291, CD 292, 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, CD3 13, CD314, CD315, CD316, CD317, CD318, CD319, CD320, CD321, CD322, CD323, CD324, CD325, CD326, CD32 7, CD328, CD329, CD330, CD331, CD332, CD333, CD334, CD335, CD336, CD337, CD338, CD339, CD340, CD344 An anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 13, selected from the group consisting of CD349, CD351, CD352, CD353, CD354, CD355, CD357, CD358, CD360, CD361, CD362, CD363, CD364, CD365, CD366, CD367, CD368, CD369, CD370, and CD371.

15. The anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 8, 10 to 11, and 13, wherein the antibody is not a bispecific antibody or a multispecific antibody.

16. The aforementioned antibody is a bispecific antibody, a. The second antigen is CD19, and optionally the antibody includes SEQ ID NO: 314; SEQ ID NO: 315; SEQ ID NO: 396; SEQ ID NO: 397; SEQ ID NO: 421 and SEQ ID NO: 422; or SEQ ID NO: 504 and SEQ ID NO:

422. b. The second antigen is Her2 (CD340), and optionally the antibody includes SEQ ID NO: 393; SEQ ID NO: 394; SEQ ID NO: 395; SEQ ID NO: 398; SEQ ID NO: 414 and SEQ ID NO: 433; SEQ ID NO: 504 and SEQ ID NO: 433; SEQ ID NO: 505 and SEQ ID NO: 433; SEQ ID NO: 414 and SEQ ID NO: 434; SEQ ID NO: 504 and SEQ ID NO: 434; or SEQ ID NO: 505 and SEQ ID NO:

434. c. The second antigen is EGFR, and optionally the antibody includes SEQ ID NO: 379; SEQ ID NO: 383; SEQ ID NO: 385; SEQ ID NO: 386; SEQ ID NO: 387; SEQ ID NO: 388; SEQ ID NO: 399; SEQ ID NO: 421 and SEQ ID NO: 437; SEQ ID NO: 504 and SEQ ID NO: 437; SEQ ID NO: 421 and SEQ ID NO: 429; SEQ ID NO: 504 and SEQ ID NO: 429; SEQ ID NO: 414 and SEQ ID NO: 431; SEQ ID NO: 414 and SEQ ID NO: 435; SEQ ID NO: 504 and SEQ ID NO: 431; or SEQ ID NO: 505 and SEQ ID NO: 435, d. The second antigen is FAPα, and optionally the antibody includes SEQ ID NO: 401; SEQ ID NO: 402; SEQ ID NO: 414 and SEQ ID NO: 415; SEQ ID NO: 504 and SEQ ID NO: 415; or SEQ ID NO: 505 and SEQ ID NO:

415. e. The second antigen is MSLN, and optionally the antibody includes SEQ ID NO: 403; SEQ ID NO: 404; SEQ ID NO: 414 and SEQ ID NO: 416; SEQ ID NO: 504 and SEQ ID NO: 416; or SEQ ID NO: 505 and SEQ ID NO:

416. f. The second antigen is PD-1, and optionally the antibody includes SEQ ID NO: 405; SEQ ID NO: 406; or SEQ ID NO: 438 and SEQ ID NO: 417, g. The second antigen is 4-1BB, and optionally the antibody includes SEQ ID NO: 407; SEQ ID NO: 408; or SEQ ID NO: 414 and SEQ ID NO: 418; SEQ ID NO: 504 and SEQ ID NO: 418; or SEQ ID NO: 505 and SEQ ID NO: 418, h. The second antigen is OX40, and optionally the antibody includes SEQ ID NO: 409; SEQ ID NO: 410; SEQ ID NO: 414 and SEQ ID NO: 419; SEQ ID NO: 504 and SEQ ID NO: 419; or SEQ ID NO: 505 and SEQ ID NO: 419, or i. The anti-Vδ1 antibody or antigen-binding fragment according to claim 14, wherein the second antigen is TIGIT, and optionally the antibody comprises SEQ ID NO: 411; SEQ ID NO: 412; or SEQ ID NO: 439 and SEQ ID NO:

420.

17. The anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 16, wherein the antibody is of the Fc-effective type.

18. A pharmaceutical composition comprising an anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 17, and a pharmaceutically acceptable diluent or carrier.

19. A pharmaceutical composition comprising an anti-Vδ1 antibody or its antigen-binding fragment according to any one of claims 1 to 17, or the pharmaceutical composition according to claim 18, for use in treating a target disease or disorder, or in modulating the immune response in a target.

20. a. Whether the disease or disorder is cancer, an infectious disease, or an inflammatory disease, or whether the subject whose immune response is regulated has cancer, an infectious disease, or an inflammatory disease, b. The pharmaceutical composition according to claim 19, wherein the modification of the immune response in the target comprises at least one selected from the group consisting of activating γδT cells, causing or enhancing γδT cell proliferation, causing or enhancing γδT cell enlargement, causing or enhancing γδT cell degranulation, causing or enhancing γδT cell killing activity, causing or enhancing γδT cell cytotoxicity, causing or enhancing γδT cell recruitment, extending the survival time of γδT cells, and enhancing γδT cell exhaustion.

21. The pharmaceutical composition according to claim 18 or 20, wherein diseased cells are killed while healthy cells are preserved.

22. A pharmaceutical composition comprising an anti-TCRδ variable portion 1 (anti-Vδ1) antibody or an antigen-binding fragment thereof according to any one of claims 1 to 17, for use in the treatment of cancer, infectious diseases, or inflammatory diseases, or the pharmaceutical composition according to claim 18.

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