Multifunctional molecules binding to TCR and uses thereof

Abstract

Provided herein are multifunctional molecules comprising a first domain that binds to a first target molecule, wherein the first target molecule is a T cell receptor alpha (TCRα) chain; and a second domain that binds to a second target molecule, wherein the second target molecule is a T cell receptor beta (TCRβ) chain, and methods of using thereof.

Description

WSGR Docket No.53676-768.601 MULTIFUNCTIONAL MOLECULES BINDING TO TCR AND USES THEREOF CROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 624,887, filed January25, 2024, which is incorporated herein by reference in its entirety. BACKGROUND

[0002] Currently available molecules designed to redirect T cells to promote tumor cell lysis for cancerimmunotherapy typically target the CD3 epsilon (CD3e) subunit of the T cell receptor (TCR). However, there are limitations to this approach. Previous studies have shown that, e.g., low doses of anti-CD3e monoclonal antibody (mAb) can cause T cell dysfunction and exert immunosuppressive effects. In addition, anti-CD3e mAbs bind to all T cells and thus activate a large number of T cells. Such non- physiological massive activation of T cells by these anti-CD3e mAbs can result in the production of proinflammatory cytokines such as IFN-gamma, IL-1-beta, IL-6, IL-10 and TNF-alpha, causing a “cytokine storm” known as the cytokine release syndrome (CRS), which is also associated with neurotoxicity (NT). Thus, there is a need for improved T cell receptor-binding molecules that redirect T cells for cancer immunotherapy. SUMMARY

[0003] In an aspect, provided herein is, inter alia, a multifunctional molecule comprising (a) a firstdomain that binds to a first target molecule, wherein the first target molecule is a T cell receptor alpha (TCRα) chain; and (b) a second domain that binds to a second target molecule, wherein the second target molecule is a T cell receptor beta (TCRβ) chain.

[0004] In some embodiments, the TCRα chain is a human TCRα .

[0005] In some embodiments, the TCRβ chain is a human TCRβ chain.

[0006] In some embodiments, the first domain is a TCRαV-binding domain that binds to a variableregion of the human TCRα chain (TCRαV).

[0007] In some embodiments, the second domain is a TCRβV-binding domain that binds to a variableregion of the human TCRβ chain (TCRβV).

[0008] In some embodiments, the multifunctional molecule comprises at least two non-contiguouspolypeptides comprising a first polypeptide chain and a second polypeptide chain; wherein the first polypeptide chain comprises a first portion of a dimerization module; wherein the second polypeptide chain comprises a second portion of the dimerization module; and wherein the first polypeptide chain and the second polypeptide chain form a dimer via association of the first portion of the dimerization module and the second portion of the dimerization module.

[0009] In some embodiments, the first domain is an antibody molecule.

[0010] In some embodiments, the second domain is an antibody molecule.WSGR Docket No.53676-768.601

[0011] In some embodiments, the antibody molecule is selected from the group consisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

[0012] In some embodiments, the first domain is a scFv or a single domain antibody.

[0013] In some embodiments, the first domain is an Fab comprising a first portion of the first domainand a second portion of the first domain; wherein the first portion of the first domain and the second portion of the first domain assemble and form the first domain of the multifunctional molecule; wherein the multifunctional molecule further comprises a polypeptide chain comprising the second portion of the first domain, wherein the polypeptide chain comprising the second portion of the first domain is non- contiguous with the first polypeptide chain and the second polypeptide chain.

[0014] In some embodiments, the first portion of the first domain comprises a heavy chain variableregion (VH) of the Fab and the second portion of the first domain comprises a light chain variable region (VL) of the Fab, or the first portion of the first domain comprises the VL of the Fab and the second portion of the first domain comprises the VH of the Fab.

[0015] In some embodiments, the second domain is a scFv or a single domain antibody.

[0016] In some embodiments, the second domain is an Fab comprising a first portion of the seconddomain and a second portion of the second domain; wherein the first portion of the second domain and the second portion of the second domain assemble and form the second domain of the multifunctional molecule; wherein the multifunctional molecule further comprises a polypeptide chain comprising the second portion of the second domain, wherein the polypeptide chain comprising the second portion of the second domain is non-contiguous with the first polypeptide chain, the second polypeptide chain, and the polypeptide chain comprising the second portion of the first domain.

[0017] In some embodiments, the first portion of the second domain comprises a heavy chain variableregion (VH) of the Fab and the second portion of the second domain comprises a light chain variable region (VL) of the Fab, or the first portion of the second domain comprises the VL of the Fab and the second portion of the second domain comprises the VH of the Fab.

[0018] In some embodiments, the first portion of the dimerization module is linked to the first domain orthe first portion of the first domain.

[0019] In some embodiments, the N-terminus of the first portion of the dimerization module is linked tothe C-terminus of the first domain or the C-terminus of the first portion of the first domain.

[0020] In some embodiments, the C-terminus of the first portion of the dimerization module is linked tothe N-terminus of the first domain or the N-terminus of the first portion of the first domain.

[0021] In some embodiments, the first portion of the dimerization module is linked to the seconddomain or the first portion of the second domain.

[0022] In some embodiments, the N-terminus of the first portion of the dimerization module is linked tothe C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the C- terminus of the first portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.WSGR Docket No.53676-768.601

[0023] In some embodiments, the C-terminus of the first portion of the dimerization module is linked tothe N-terminus of the first domain or the N-terminus of the first portion of the first domain, and the N- terminus of the first portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

[0024] In some embodiments, the second portion of the dimerization module is linked to the seconddomain or the first portion of the second domain.

[0025] In some embodiments, the N-terminus of the second portion of the dimerization module is linkedto the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

[0026] In some embodiments, the C-terminus of the second portion of the dimerization module is linkedto the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

[0027] In some embodiments, the N-terminus of the first portion of a dimerization module is linked tothe C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the N- terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

[0028] In some embodiments, the N-terminus of the first portion of the dimerization module is linked tothe C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the C- terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

[0029] In some embodiments, the C-terminus of the first portion of the dimerization module is linked tothe N-terminus of the first domain or the N-terminus of the first portion of the first domain, and the C- terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

[0030] In some embodiments, the C-terminus of the first portion of the dimerization module is linked tothe N-terminus of the first domain or the N-terminus of the first portion of the first domain, and the N- terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

[0031] In some embodiments, the multifunctional molecule comprises at least three non-contiguouspolypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to a first portion of the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module; and (iii) a polypeptide chain comprising a second portion of the first domain, and wherein the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

[0032] In some embodiments, the multifunctional molecule comprises at least three non-contiguouspolypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module; and (iii) a polypeptide chain comprising a second portion of the second domain, and wherein a first portion of the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.WSGR Docket No.53676-768.601

[0033] In some embodiments, the multifunctional molecule comprises at least four non-contiguouspolypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to a first portion of the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module;(iii) a polypeptide chain comprising a second portion of the first domain; and (iv) a polypeptide chain comprising a second portion of the second domain, and wherein a first portion of the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

[0034] In some embodiments, the multifunctional molecule comprises at least two non-contiguouspolypeptide chains, wherein: the first polypeptide chain comprising the first portion of the dimerization module linked to the first domain; and the second polypeptide chain comprising the second portion of the dimerization module; and wherein the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

[0035] In some embodiments, the multifunctional molecule further comprises at least one cytokine or afunctional fragment or variant thereof.

[0036] In some embodiments, the at least one cytokine or a functional fragment or variant thereof islinked to the first portion of the dimerization module, the second portion of the dimerization module, or any combination thereof.

[0037] In some embodiments, the at least one cytokine or a functional fragment or variant thereof islinked to the N-terminus of the first portion of the dimerization module, the C-terminus of the first portion of the dimerization module, the N-terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof.

[0038] In some embodiments, the at least one cytokine or a functional fragment or variant thereof islinked to the first domain, the first portion of the first domain, the second portion of the first domain, the second domain, the first portion of the second domain, the second portion of the second domain, or any combination thereof.

[0039] In some embodiments, the at least one cytokine or a functional fragment or variant thereof islinked to the N-terminus of the first domain, the C-terminus of the first domain, the N-terminus of the first portion of the first domain, the C-terminus of the first portion of the first domain, the N-terminus of the second portion of the first domain, the C-terminus of the second portion of the first domain, the N- terminus of the second domain, the C-terminus of the second domain, the N-terminus of the first portion of the second domain, the C-terminus of the first portion of the second domain, the N-terminus of the second portion of the second domain, the C-terminus of the second portion of the second domain, or any combination thereof.

[0040] In some embodiments, the at least one cytokine or a functional fragment or variant thereof isselected from the group consisting of interleukin-2 (IL-2) or functional variant thereof, interleukin-7 (IL- 7) or functional variant thereof, interleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL- 15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL-WSGR Docket No.53676-768.601 21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

[0041] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises interleukin-2 (IL-2) or functional variant thereof.

[0042] In some embodiments, the interleukin-2 (IL-2) or functional variant thereof comprises asequence with at least 70% sequence identity to the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

[0043] In some embodiments, the interleukin-2 (IL-2) or functional variant thereof comprises thesequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

[0044] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises interleukin-15 (IL-15) or functional variant thereof.

[0045] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof comprises asequence with at least 70% sequence identity to the sequence of SEQ ID NO: 2170.

[0046] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof comprises thesequence of SEQ ID NO: 2170.

[0047] In some embodiments, the at least one cytokine or a functional fragment or variant thereoffurther comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

[0048] In some embodiments, the IL15Ralpha dimerizing domain comprises an IL-15 receptor alphasushi domain or functional variant thereof.

[0049] In some embodiments, the IL15Ralpha dimerizing domain comprises a sequence with at least70% sequence identity to the sequence of SEQ ID NO: 3472.

[0050] The multifunctional molecule of any one of claims 45-47, wherein the IL15Ralpha dimerizingdomain comprises the sequence of SEQ ID NO: 3472.

[0051] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof is covalentlylinked to the IL15Ralpha dimerizing domain via a linker.

[0052] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof is covalentlylinked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

[0053] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises a sequence with at least 70% sequence identity to the sequence of SEQ ID NO: 3474.

[0054] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises the sequence of SEQ ID NO: 3474.

[0055] In some embodiments, the multifunctional molecule further comprises a linker between the firstdomain or the first portion of the first domain and the first portion of the dimerization module.

[0056] In some embodiments, the multifunctional molecule further comprises a linker between thesecond domain or the first portion of the second domain and the first portion of a dimerization module.

[0057] In some embodiments, the multifunctional molecule further comprises a linker between thesecond domain or the first portion of the second domain and the second portion of a dimerization module.WSGR Docket No.53676-768.601

[0058] In some embodiments, the multifunctional molecule further comprises a linker between the atleast one cytokine or a functional fragment or variant thereof and the first portion of the dimerization module.

[0059] In some embodiments, the multifunctional molecule further comprises a linker between the atleast one cytokine or a functional fragment or variant thereof and the first portion of the dimerization module of the first polypeptide chain.

[0060] In some embodiments, the multifunctional molecule further comprises a linker between the atleast one cytokine or a functional fragment or variant thereof and the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second portion of the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the second domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second portion of the second domain, or any combination thereof.

[0061] In some embodiments, the multifunctional molecule further comprises a tumor-targeting moiety.

[0062] In some embodiments, the tumor-targeting moiety binds to a cancer antigen.

[0063] In some embodiments, the tumor-targeting moiety binds to a cancer antigen selected from thegroup consisting of CD19, CD123, CD22, CD30, CD171, CS-1, C-type lectin-like molecule-1, CD33, epidermal growth factor receptor variant III (EGFRvIII), ganglioside G2 (GD2), ganglioside GD3, TNF receptor family member B cell maturation (BCMA), Tn antigen ((Tn Ag) or (GalNAcα-Ser / Thr)), prostate-specific membrane antigen (PSMA), Receptor tyrosine kinase-like orphan receptor 1 (ROR1), Fms-Like Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD117), Interleukin-13 receptor subunit alpha-2, mesothelin, Interleukin 11 receptor alpha (IL-11Ra), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage- specific embryonic antigen-4 (SSEA-4), CD20, Folate receptor alpha, Receptor tyrosine-protein kinase ERBB2 (Her2 / neu), Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion molecule (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gp100) / pmel17, oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), tyrosinase, ephrin type-A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma-associated antigen (HMWMAA), o-acetyl-GD2 ganglioside (OAcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related (TEM7R), claudin 6 (CLDN6), thyroid stimulatingWSGR Docket No.53676-768.601 hormone receptor (TSHR), G protein-coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, CD179a, anaplastic lymphoma kinase (ALK), Polysialic acid, placenta-specific 1 (PLAC1), hexasaccharide portion of globoH glycoceramide (GloboH), mammary gland differentiation antigen (NY-BR-1), uroplakin 2 (UPK2), Hepatitis A virus cellular receptor 1 (HAVCR1), adrenoceptor beta 3 (ADRB3), pannexin 3 (PANX3), G protein-coupled receptor 20 (GPR20), lymphocyte antigen 6 complex, locus K 9 (LY6K), Olfactory receptor 51E2 (OR51E2), TCR Gamma Alternate Reading Frame Protein (TARP), Wilms tumor protein (WT1), Cancer / testis antigen 1 (NY-ESO-1) / LAGE-1, Cancer / testis antigen 2 (LAGE-1a), Melanoma-associated antigen 1 (MAGE-A1), ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML), sperm protein 17 (SPA17), X Antigen Family, Member 1A (XAGE1), angiopoietin-binding cell surface receptor 2 (Tie 2), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), Fos-related antigen 1, tumor protein p53 (p53), p53 mutant, prostein, survivin, telomerase, prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1, Rat sarcoma (Ras) mutant, human Telomerase reverse transcriptase (hTERT), sarcoma translocation breakpoints, melanoma inhibitor of apoptosis (ML-IAP), ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene), N-Acetyl glucosaminyl-transferase V (NA17), paired box protein Pax-3 (PAX3), Androgen receptor, Cyclin B1, v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN), Ras Homolog Family Member C (RhoC), Tyrosinase-related protein 2 (TRP-2), Cytochrome P4501B1 (CYP1B1), CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3), Paired box protein Pax-5 (PAX5), proacrosin binding protein sp32 (OY-TES1), lymphocyte-specific protein tyrosine kinase (LCK), A kinase anchor protein 4 (AKAP-4), synovial sarcoma, X breakpoint 2 (SSX2), Receptor for Advanced Glycation Endproducts (RAGE-1), renal ubiquitous 1 (RU1), renal ubiquitous 2 (RU2), legumain, human papilloma virus E6 (HPV E6), human papilloma virus E7 (HPV E7), intestinal carboxyl esterase, heat shock protein 70-2 mutated (mut hsp70-2), CD79a, CD79b, CD72, Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), Fc fragment of IgA receptor (FCAR or CD89), Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), CD300 molecule-like family member f (CD300LF), C-type lectin domain family 12 member A (CLEC12A), bone marrow stromal cell antigen 2 (BST2), EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2), lymphocyte antigen 75 (LY75), Glypican-3 (GPC3), Fc receptor-like 5 (FCRL5), FcRH5, PDL1, CD47, prostate specific membrane antigen (PMSA), prostate- specific antigen (PSA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium- activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, SAP-1, PRAME, Melan-A / MART-1, TRP1 / gp75, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, Ras, TGF-Β receptor, TGF-β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, gangliosides, MART-2, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, L1-CAM, gpA33, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, TGF-beta, hyaluronic acid, collagen, tenascin C, tenascin W, and immunoglobulin lambda-like polypeptide 1 (IGLL1).WSGR Docket No.53676-768.601

[0064] In some embodiments, the tumor-targeting moiety is an antibody molecule.

[0065] In some embodiments, the antibody molecule is selected from the group consisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

[0066] In some embodiments, the multifunctional molecule further comprises a stromal modifyingmoiety.

[0067] In some embodiments, the multifunctional molecule further comprises an immune cell engager.

[0068] In some embodiments, the immune cell engager is selected from the group consisting of a T cellengager, an NK cell engager, a B cell engager, a dendritic cell engager, the macrophage cell engager, and any combination thereof.

[0069] In some embodiments, immune cell engager is an antibody molecule.

[0070] In some embodiments, the antibody molecule is selected from the group consisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

[0071] In some embodiments, the first portion of the dimerization module of the first polypeptide chainis a first Fc region or variant thereof.

[0072] In some embodiments, the second portion of the dimerization module of the second polypeptidechain is a second Fc region or variant thereof.

[0073] In some embodiments, the first portion of the dimerization module, the second portion of thedimerization module, or a combination thereof is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ Fc region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

[0074] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises an engineered Fc interface with one or more of: a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, wherein dimerization of the first Fc region and the second Fc region is enhanced as indicated by a greater ratio of heteromultimer:homomultimer forms relative to dimerization of Fc regions without the engineered interface.

[0075] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises a Cys at position 349, a Ser at position 366, an Ala at position 368, a Val at position 407, a Cys at position 354, a Trp at position 366, or any combination thereof in a heavy chain constant region according to EU Numbering.

[0076] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises (i) a Cys at position 349, a Ser at position 366, an Ala at position 368, and a Val at position 407 in a heavy chain constant region according to EU Numbering; (ii) a Cys at position 354 and a Trp at position 366 in a heavy chain constant region according to EU Numbering; or (iii) a combination thereof.WSGR Docket No.53676-768.601

[0077] In some embodiments, (i) the first Fc region comprises: (a) a Cys at position 349 in a heavychain constant region according to EU Numbering, (b) a Ser at position 366 in a heavy chain constant region according to EU Numbering, (c) an Ala at position 368 in a heavy chain constant region according to EU Numbering, and (d) a Val at position 407 in a heavy chain constant region according to EU Numbering; and (ii) the second Fc region comprises: (a) a Cys at position 354 in a heavy chain constant region according to EU Numbering, and (b) a Trp at position 366 in a heavy chain constant region according to EU Numbering.

[0078] In some embodiments, (i) the first Fc region comprises:(a) a Cys at position 354 in a heavy chainconstant region according to EU Numbering, and (b) a Trp at position 366 in a heavy chain constant region according to EU Numbering(ii) the second Fc region comprises: (a) a Cys at position 349 in a heavy chain constant region according to EU Numbering, (b) a Ser at position 366 in a heavy chain constant region according to EU Numbering, (c) an Ala at position 368 in a heavy chain constant region according to EU Numbering, and (d) a Val at position 407 in a heavy chain constant region according to EU Numbering.

[0079] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprise one or more mutations that result in reduced or ablated affinity for at least one Fc receptor relative to a Fc region without the one or more mutations.

[0080] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

[0081] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises a sequence having at least 70% sequence identity to the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 3901, SEQ ID NO: 3645, SEQ ID NO: 3902, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO: 3903, SEQ ID NO:206, SEQ ID NO: 207, SEQ ID NO: 3904, SEQ ID NO: 3452, SEQ ID NO: 3447, or SEQ ID NO: 3453.

[0082] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 3901, SEQ ID NO: 3645, SEQ ID NO: 3902, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO: 3903, SEQ ID NO:206, SEQ ID NO: 207, SEQ ID NO: 3904, SEQ ID NO: 3452, SEQ ID NO: 3447, or SEQ ID NO: 3453.

[0083] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises a sequence having at least 70% sequence identity to the sequence of SEQ ID NO:206, SEQ ID NO: 3447, or SEQ ID NO: 3453.

[0084] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises the sequence of SEQ ID NO:206, SEQ ID NO: 3447, or SEQ ID NO: 3453.

[0085] In some embodiments, the multifunctional the molecule comprises the following configuration:A-[first portion of dimerization module]-C, B-[second portion of dimerization module]-D, wherein:WSGR Docket No.53676-768.601 (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first domain; (iii) the second domain, (iv) the at least one cytokine or a functional fragment or variant thereof, (v) the tumor targeting moiety, (vi) the stromal modifying moiety; or (vii) the immune cell engager; wherein at least one of A or C are the first domain, and wherein when A is the first domain, at least one of B, C, or D is the second domain, and when C is the first domain, at least one of A, B, or D is the second domain.

[0086] In some embodiments, the multifunctional molecule is not immobilized to a solid-phase.

[0087] In some embodiments, the multifunctional molecule is not immobilized to a solid-phase.

[0088] In some embodiments, the second domain binds to one or more of a TCRβV subfamily selectedfrom the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 subfamily, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 subfamily, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

[0089] In some embodiments, the second domain binds to one or more of a TCRβV subfamily selectedfrom the group consisting of: TCRβ V1 subfamily comprising TCRβ V1*01; TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02, and TCRβ V2*03; TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04; TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04, TCRβ V5-5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5-8*01, and TCRβ V5-8*02; TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6- 3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6- 6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6-9*01; TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7-3*03, TCRβ V7-3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7-7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7-8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7-9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7-9*06, and TCRβ V7-9*07; TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02; TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-WSGR Docket No.53676-768.601 1*02, and TCRβ V9-1*03; TCRβ V10 subfamily comprising one or more selected from TCRβ V10- 1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10-3*02, TCRβ V10-3*03, and TCRβ V10-3*04; TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02, TCRβ V11-2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11- 3*03, and TCRβ V11-3*04; TCRβ V12 subfamily comprising one or more selected from TCRβ V12- 3*01, TCRβ V12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01; TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβ V13*02; TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβ V14*02; TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβ V15*02, and TCRβ V15*03; TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβ V16*02, and TCRβ V16*03; TCRβ V17 subfamily comprising TCRβ V17*01;TCRβ V18 subfamily comprising TCRβ V18*01; TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβ V19*02, and TCRβ V19*03; TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβ V20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07;TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβ V21-1*02;TCRβ V22 subfamily comprising TCRβ V22-1*01;TCRβ V23 subfamily comprising TCRβ V23-1*01;TCRβ V24 subfamily comprising TCRβ V24-1*01;TCRβ V25 subfamily comprising TCRβ V25-1*01;TCRβ V26 subfamily comprising TCRβ V26-1*01;TCRβ V27 subfamily comprising TCRβ V27*01;TCRβ V28 subfamily comprising TCRβ V28*01;TCRβ V29 subfamily comprising one or more selected from TCRβ V29- 1*01, TCRβ V29-1*02, and TCRβ V29-1*03; and TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβ V30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

[0090] In some embodiments, the second domain binds to TCRβ V4 subfamily, TCRβ V6 subfamily,TCRβ V20 subfamily, or TCRβ V25 subfamily.

[0091] In some embodiments, the second domain binds to TCRΒ V4-1, TCRΒ V6-1, TCRΒ V6-2,TCRΒ V6-3, TCRΒ V6-4, TCRΒ V6-5, TCRΒ V6-6, TCRΒ V6-7, TCRΒ V6-8, TCRΒ V6-9, TCRΒ V20-1, or TCRΒ V25-1.

[0092] In some embodiments, the multifunctional molecule comprises a single TCRβV-binding domain.

[0093] In some embodiments, the first domain binds to one or more of a TCRαV subfamily selectedfrom the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.WSGR Docket No.53676-768.601

[0094] In some embodiments, the first domain binds to one or more of a TCRαV subfamily selectedfrom the group consisting of: TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02;TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02;TCRα V3 subfamily comprising TCRα V3*01;TCRα V4 subfamily comprising TCRα V4*01;TCRα V5 subfamily comprising TCRα V5*01TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06;TCRα V7 subfamily comprising TCRα V7*01;TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8- 2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8-4*01, TCRα V8-4*02, TCRα V8- 4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8- 6*02, and TCRα V8-7*01; TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; TCRα V10 subfamily comprising TCRα V10*01;TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12- 3*02;TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02;TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04;TCRα V16 subfamily comprising TCRα V16*01;TCRα V17 subfamily comprising TCRα V17*01;TCRα V18 subfamily comprising TCRα V18*01;TCRα V19 subfamily comprising TCRα V19*01TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04;TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02;TCRα V22 subfamily comprising TCRα V22*01;TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04;TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02;TCRα V25 subfamily comprising TCRα V25*01;TCRα V26 subfamily comprising one or more selected from TCRα V26- 1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02;TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03;TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02;TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04;TCRα V34 subfamily comprising TCRα V34*01;TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02;TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04;TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38- 1*04, and TCRα V38-2*01;TCRα V39 subfamily comprising TCRα V39*01;TCRα V40 subfamily comprising TCRα V40*01; and TCRα V41 subfamily comprising TCRα V41*01.

[0095] In some embodiments, the second domain binds to TCRα V12 subfamily, TCRα V13 subfamily,TCRα V19 subfamily, TCRα V21 subfamily, or TCRα V30 subfamily.WSGR Docket No.53676-768.601

[0096] In some embodiments, the second domain binds to TCRα V1 subfamily, TCRα V10 subfamily,TCRα V17 subfamily, or TCRα V19 subfamily.

[0097] In some embodiments, the second domain binds to TCRα V1-2, TCRα V10, or TCRα V17.

[0098] In some embodiments, the first domain and the second domain bind to: TCRα V1 and TCRβ V6,respectively; TCRα V1 and TCRβ V20, respectively; TCRα V10 and TCRβ V25, respectively; TCRα V1 and TCRβ V4, respectively; TCRα V17 and TCRβ V4, respectively; or TCRα V17 and TCRβ V6, respectively.

[0099] In some embodiments, the first domain and the second domain bind to: TCRα V1-2 and TCRβV6-1, respectively; TCRα V1-2 and TCRβ V6-2, respectively; TCRα V1-2 and TCRβ V6-3, respectively; TCRα V1-2 and TCRβ V6-4, respectively; TCRα V1-2 and TCRβ V6-5, respectively; TCRα V1-2 and TCRβ V6-6, respectively; TCRα V1-2 and TCRβ V6-7, respectively; TCRα V1-2 and TCRβ V6-8, respectively; TCRα V1-2 and TCRβ V6-9, respectively; TCRα V1-2 and TCRβ V20-1, respectively; TCRα V10 and TCRβ V25-1, respectively; TCRα V1-2 and TCRβ V4-1, respectively; TCRα V17 and TCRβ V4-1, respectively; or TCRα V17 and TCRβ V6-2, respectively.

[0100] In some embodiments, the first domain comprises an antibody molecule that binds to the TCRαVcomprising (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Table 22; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Table 22; or (iii) a combination thereof.

[0101] In some embodiments, the first domain comprises an antibody molecule that binds to theTCRαV comprising (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

[0102] In some embodiments, the first domain comprises an antibody molecule that binds to the TCRαVcomprising (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Table 22; (i) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

[0103] In some embodiments, the first domain comprises an antibody molecule that binds to the TCRαVcomprising (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

[0104] In some embodiments, the first domain comprises any one of antibody molecules that binds tothe TCRαV listed in Table 22.

[0105] In some embodiments, the second domain comprises an antibody molecule that binds to theTCRβV comprising (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VLWSGR Docket No.53676-768.601 comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

[0106] In some embodiments, the second domain comprises an antibody molecule that binds to theTCRβV comprising (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

[0107] In some embodiments, the second domain comprises an antibody molecule that binds to theTCRβV comprising (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (i) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

[0108] In some embodiments, the second domain comprises an antibody molecule that binds to theTCRβV comprising (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

[0109] In some embodiments, the second domain comprises any one of antibody molecules that binds tothe TCRβV listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26.

[0110] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises one or more mutations listed in tables 4 and 14 according to EU numbering.

[0111] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises a sequence having at least 70% sequence identity to any one of the heavy chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0112] In some embodiments, the first Fc region, the second Fc region, or a combination thereofcomprises any one of the heavy chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0113] In some embodiments, the multifunctional molecule further comprises one or moreimmunoglobulin light chain constant regions.

[0114] In some embodiments, the one or more immunoglobulin light chain constant region is linked tothe first domain, the first portion of the first domain, or the second portion of the first domain.

[0115] In some embodiments, the one or more immunoglobulin light chain constant region is linked tothe second domain, the first portion of the second domain, or the second portion of the second domain.

[0116] In some embodiments, the one or more immunoglobulin light chain constant region is linked tothe at least one cytokine or a functional fragment or variant thereof.WSGR Docket No.53676-768.601

[0117] In some embodiments, the one or more immunoglobulin light chain constant region comprises akappa light chain constant region, a lambda light chain constant region, or a combination thereof.

[0118] In some embodiments, the one or more immunoglobulin light chain constant region comprises asequence having at least 70% sequence identity to any one of the light chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0119] In some embodiments, the one or more immunoglobulin light chain constant region comprisesany one of the light chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0120] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises a sequence having at least 70% sequence identity to any one of the cytokine sequences listed in Tables 10C, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0121] In some embodiments, the at least one cytokine or a functional fragment or variant thereofcomprises any one of the cytokine sequences listed in Tables 10C, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

[0122] In some embodiments, the tumor-targeting moiety comprises a sequence having at least 70%sequence identity to any one of the antibody sequences listed in Table 24.

[0123] In some embodiments, the tumor-targeting moiety comprises any one of the antibody sequenceslisted in Table 24.

[0124] In some embodiments, the multifunctional molecule does not comprise an anti-CD3 bindingdomain.

[0125] In some embodiments, the multifunctional molecule is a polypeptide molecule.

[0126] In some embodiments, the multifunctional molecule is a multispecific molecule.

[0127] In another aspect, provided herein is a polynucleotide comprising a sequence encoding themultifunctional molecule as described herein.

[0128] In some embodiments, the polynucleotide is an isolated nucleic acid molecule.

[0129] In another aspect, provided herein is a vector comprising the polynucleotide as described herein.

[0130] In another aspect, provided herein is a cell comprising the multifunctional molecule as describedherein, the polynucleotide as described herein, or the vector as described herein.

[0131] In another aspect, provided herein is a method of making the multifunctional molecule asdescribed herein comprising: culturing a cell comprising the polynucleotide as described herein or the vector as described herein under conditions suitable for expression of the multifunctional molecule. Also provided herein is a method of making the multifunctional molecule as described herein comprising: culturing the cell as described herein under conditions suitable for expression of the multifunctional molecule.

[0132] In another aspect, provided herein is a composition comprising the multifunctional molecule asdescribed herein.

[0133] In another aspect, provided herein is a pharmaceutical composition comprising themultifunctional molecule as described herein, the polynucleotide as described herein, the vector asWSGR Docket No.53676-768.601 described herein, the cell as described herein, or the composition as described herein, and a pharmaceutically acceptable carrier, excipient, or diluent.

[0134] In another aspect, provided herein is a method of treating a condition or disease in a subject inneed thereof comprising administering to the subject a therapeutically effective amount of the multifunctional molecule as described herein, the polynucleotide as described herein, the vector as described herein, the cell as described herein, the composition as described herein, the pharmaceutical composition as described herein, or a combination thereof, wherein the administering is effective to treat the condition or disease in the subject.

[0135] In some embodiments, the condition or disease is cancer.

[0136] In some embodiments, the cancer is a solid tumor, a hematological cancer, a metastatic cancer, asoft tissue tumor, or a combination thereof.

[0137] In some embodiments, wherein the cancer is the solid tumor, and wherein the solid tumor isselected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

[0138] In some embodiments, the cancer is the hematological cancer, and wherein the hematologicalcancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T- cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

[0139] In some embodiments, the Non-Hodgkin’s lymphoma is selected from the group consisting of Bcell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

[0140] In some embodiments, the T-cell lymphoma is peripheral T-cell lymphoma.

[0141] In some embodiments, the cancer is characterized by a cancer antigen present on the cancer.

[0142] In some embodiments, the cancer antigen present on the cancer is a tumor antigen, a stromalantigen, or a hematological antigen.

[0143] In some embodiments, the cancer antigen is selected from the group consisting of CD19, CD123,CD22, CD30, CD171, CS-1, C-type lectin-like molecule-1, CD33, epidermal growth factor receptor variant III (EGFRvIII), ganglioside G2 (GD2), ganglioside GD3, TNF receptor family member B cell maturation (BCMA), Tn antigen ((Tn Ag) or (GalNAcα-Ser / Thr)), prostate-specific membrane antigen (PSMA), Receptor tyrosine kinase-like orphan receptor 1 (ROR1), Fms-Like Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD117), Interleukin-13 receptor subunit alpha-2, mesothelin, Interleukin 11 receptor alpha (IL-11Ra), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage-specific embryonic antigen-4 (SSEA- 4), CD20, Folate receptor alpha, Receptor tyrosine-protein kinase ERBB2 (Her2 / neu), Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion moleculeWSGR Docket No.53676-768.601 (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gp100) / pmel17, oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), tyrosinase, ephrin type-A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma-associated antigen (HMWMAA), o-acetyl-GD2 ganglioside (OAcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related (TEM7R), claudin 6 (CLDN6), thyroid stimulating hormone receptor (TSHR), G protein- coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, CD179a, anaplastic lymphoma kinase (ALK), Polysialic acid, placenta-specific 1 (PLAC1), hexasaccharide portion of globoH glycoceramide (GloboH), mammary gland differentiation antigen (NY-BR-1), uroplakin 2 (UPK2), Hepatitis A virus cellular receptor 1 (HAVCR1), adrenoceptor beta 3 (ADRB3), pannexin 3 (PANX3), G protein-coupled receptor 20 (GPR20), lymphocyte antigen 6 complex, locus K 9 (LY6K), Olfactory receptor 51E2 (OR51E2), TCR Gamma Alternate Reading Frame Protein (TARP), Wilms tumor protein (WT1), Cancer / testis antigen 1 (NY-ESO-1) / LAGE-1, Cancer / testis antigen 2 (LAGE-1a), Melanoma-associated antigen 1 (MAGE-A1), ETS translocation- variant gene 6, located on chromosome 12p (ETV6-AML), sperm protein 17 (SPA17), X Antigen Family, Member 1A (XAGE1), angiopoietin-binding cell surface receptor 2 (Tie 2), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), Fos-related antigen 1, tumor protein p53 (p53), p53 mutant, prostein, survivin, telomerase, prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1, Rat sarcoma (Ras) mutant, human Telomerase reverse transcriptase (hTERT), sarcoma translocation breakpoints, melanoma inhibitor of apoptosis (ML-IAP), ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene), N-Acetyl glucosaminyl- transferase V (NA17), paired box protein Pax-3 (PAX3), Androgen receptor, Cyclin B1, v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN), Ras Homolog Family Member C (RhoC), Tyrosinase-related protein 2 (TRP-2), Cytochrome P4501B1 (CYP1B1), CCCTC- Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3), Paired box protein Pax-5 (PAX5), proacrosin binding protein sp32 (OY-TES1), lymphocyte- specific protein tyrosine kinase (LCK), A kinase anchor protein 4 (AKAP-4), synovial sarcoma, X breakpoint 2 (SSX2), Receptor for Advanced Glycation Endproducts (RAGE-1), renal ubiquitous 1 (RU1), renal ubiquitous 2 (RU2), legumain, human papilloma virus E6 (HPV E6), human papilloma virus E7 (HPV E7), intestinal carboxyl esterase, heat shock protein 70-2 mutated (mut hsp70-2), CD79a, CD79b, CD72, Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), Fc fragment of IgA receptor (FCAR or CD89), Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), CD300 molecule-like family member f (CD300LF), C-type lectin domain family 12 member A (CLEC12A), bone marrow stromal cell antigen 2 (BST2), EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2), lymphocyte antigen 75 (LY75), Glypican-3 (GPC3), Fc receptor-like 5WSGR Docket No.53676-768.601 (FCRL5), FcRH5, PDL1, CD47, prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, SAP-1, PRAME, Melan-A / MART-1, TRP1 / gp75, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, Ras, TGF-Β receptor, TGF-β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, gangliosides, MART-2, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, L1-CAM, gpA33, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, TGF-beta, hyaluronic acid, collagen, tenascin C, tenascin W, and immunoglobulin lambda-like polypeptide 1 (IGLL1).

[0144] In some embodiments, the method as described herein further comprises administering a secondtherapeutic agent or therapy to the subject.

[0145] In some embodiments, the second therapeutic agent or therapy comprises a chemotherapeuticagent, a biologic agent, a hormonal therapy, radiation, or surgery.

[0146] In some embodiments, the second therapeutic agent or therapy is administered in combinationwith the multifunctional molecule as described herein, the polynucleotide as described herein, the vector as described herein, the cell as described herein, the composition as described herein, or the pharmaceutical composition as described herein, sequentially, simultaneously, or concurrently.

[0147] In another aspect, provided herein is a method of expansion of a subset of T cells in a T cellpopulation comprising contacting the T cell population with the multifunctional molecule as described herein or the composition as described herein, thereby expanding the subset of T cells in the T cell population.

[0148] In some embodiments, the T cell population is a human T cell population.

[0149] In some embodiments, the subset of T cells are a subset of human T cells.

[0150] In some embodiments, the subset of T cells express a T cell receptor (TCR) comprising a TCRβchain that comprises the TCRβV to which the second domain binds.

[0151] In some embodiments, the subset of T cells express a TCR comprising a TCRα chain thatcomprises the TCRαV to which the first domain binds.

[0152] In some embodiments, the TCR is a human TCR.

[0153] In some embodiments, the multispecific molecule is an agonist of the TCR.

[0154] In some embodiments, the T cell population is an in vivo T cell population.

[0155] In some embodiments, the T cell population is an ex vivo T cell population.INCORPORATION BY REFERENCE

[0156] All publications, patents, and patent applications mentioned in this specification are hereinincorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0157] The novel features of the invention are set forth with particularity in the appended claims. Abetter understanding of the features and advantages of the present invention will be obtained by referenceWSGR Docket No.53676-768.601 to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0158] FIGs. 1A-1T depict exemplary embodiments of the multifunctional molecules as describedherein. In some embodiments, exemplary multifunctional molecules as described herein comprise a first domain as described herein and a second domain as described herein, and further comprises at least one cytokine or a functional fragment or variant thereof (e.g., IL2 or a functional fragment or variant thereof). FIGs.1A, 1B and 1C depict exemplary embodiments of the multifunctional molecules as described herein containing multiple, e.g., two, molecules of a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof), linked to a first domain as described herein and a second domain as described herein. FIGs.1D, 1E and 1F depict exemplary embodiments of the multifunctional molecules as described herein containing a single molecule of a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof) linked to a first domain as described herein or a second domain as described herein. FIGs.1G, 1H, 1I, and 1J depict exemplary embodiments of the multifunctional molecules as described herein containing at least one molecule of a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof) linked to the dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a first portion of the dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a second portion of the dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a first portion of the dimerization module and a second portion of the dimerization module. FIGs.1K, 1L and 1M depict exemplary embodiments of the multifunctional molecules as described herein containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation, and multiple, e.g., two, molecules of a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof) linked to a first domain as described herein and a second domain as described herein. FIGs.1N, 1O and 1P depict exemplary embodiments of the multifunctional molecules as described herein containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation (Knob-in-hole), and a single molecule of a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof) linked to a first domain as described herein or a second domain as described herein. FIGs.1Q, 1R, 1S and 1T depict exemplary embodiments of the multifunctional molecules as described herein containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation (Knob-in-hole), and a cytokine or a functional fragment or variant thereof (e.g., interleukin-2 (IL-2) or a functional fragment or variant thereof) linked to the exemplary dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a first portion of the dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a second portion of the dimerization module. In some embodiments, the cytokine or a functional fragment or variant thereof is linked to a first portion of the dimerization module and a second portion of the dimerization module. In some embodiments, a firstWSGR Docket No.53676-768.601 domain as described herein binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, the exemplary multifunctional molecules as described herein do not comprise at least one cytokine or a functional fragment or variant thereof.

[0159] FIGs. 2A-2B shows the alignment of the Antibody A source mouse VH and VL framework 1,CDR 1, framework 2, CDR 2, framework 3, CDR3, and framework 4 regions with their respective humanized sequences. Kabat CDRs are shown in bold, Chothia CDRs are shown in italics, and combined CDRs are shown in boxes. The framework positions that were back mutated are double underlined. FIG. 2A shows VH sequences for murine Antibody A (SEQ ID NO: 1) and humanized Antibody A-H (SEQ ID NO: 9). FIG.2B shows VL sequences for murine Antibody A (SEQ ID NO: 2) and humanized Antibody A-H (SEQ ID NO: 10 and SEQ ID NO: 11).

[0160] FIGs. 3A-3B shows the alignment of the Antibody B source mouse VH and VL framework 1,CDR 1, framework 2, CDR 2, framework 3, CDR3, and framework 4 regions with their respective humanized sequences. Kabat CDRs are shown in bold, Chothia CDRs are shown in italics, and combined CDRs are shown in boxes. The framework positions that were back mutated are double underlined. FIG. 3A shows the VH sequence for murine Antibody B (SEQ ID NO: 15) and humanized VH sequences B- H.1A to B-H.1C (SEQ ID NOs: 23-25). FIG.3B shows the VL sequence for murine Antibody B (SEQ ID NO: 16) and humanized VL sequences B-H.1D to B-H.1H (SEQ ID NOs: 26-30).

[0161] FIG. 4 depicts the phylogenetic tree of TCRBV gene family and subfamilies with correspondingantibodies mapped. Subfamily identities are as follows: Subfamily A: TCRβ V6; Subfamily B: TCRβ V10; Subfamily C: TCRβ V12; Subfamily D: TCRβ V5; Subfamily E: TCRβ V7; Subfamily F: TCRβ V11; Subfamily G: TCRβ V14; Subfamily H: TCRβ V16; Subfamily I:TCRβ V18; Subfamily J:TCRβ V9; Subfamily K: TCRβ V13; Subfamily L: TCRβ V4; Subfamily M:TCRβ V3; Subfamily N:TCRβ V2; Subfamily O:TCRβ V15; Subfamily P: TCRβ V30; Subfamily Q: TCRβ V19; Subfamily R:TCRβ V27; Subfamily S:TCRβ V28; Subfamily T: TCRβ V24; Subfamily U: TCRβ V20; Subfamily V: TCRβ V25; and Subfamily W:TCRβ V29 subfamily. Subfamily members are described in detail herein in the Section titled “TCR beta V (TCRβV)”.

[0162] FIGs. 5A-5C show human CD3+ T cells activated by anti-TCR Vβ13.1 antibody (A-H.1) for 6-days. Human CD3+ T cells were isolated using magnetic-bead separation (negative selection) and activated with immobilized (plate-coated) anti-TCR Vβ13.1 (A-H.1) or anti-CD3^ (OKT3) antibodies at 100 nM for 6 days. FIG.5A shows two scatter plots (left: activated with OKT3; and right: activated withWSGR Docket No.53676-768.601 A-H.1) of expanded T cells assessed for TCR Vβ13.1 surface expression using anti-TCR Vβ13.1 (A-H.1) followed by a secondary fluorochrome- conjugated antibody for flow cytometry analysis. FIG.5B shows percentage (%) of TCR Vβ13.1 positive T cells activated by anti-TCR Vβ13.1 (A-H.1) or anti-CD3e (OKT3) plotted against total T cells (CD3+). FIG.5C shows relative cell count acquired by counting the number of events in each T cell subset gate (CD3 or TCR Vβ13.1) for 20 seconds at a constant rate of 60μl / min. Data shown as mean value from 3 donors.

[0163] FIGs. 6A-6B show cytolytic activity of human CD3+ T cells activated by anti-TCR Vβ13.1antibody (A-H.1) against transformed cell line RPMI 8226. FIG.6A depicts target cell lysis of human CD3+ T cells activated with A-H.1or OKT3. Human CD3+ T cells were isolated using magnetic-bead separation (negative selection) and activated with immobilized (plate-coated) A-H.1 or OKT3 at the indicated concentrations for 4 days prior to co-culture with RPMI 8226 cells at a (E:T) ratio of 5:1 for 2 days. Samples were next analyzed for cell lysis of RPMI 8226 cells by FACS staining for CFSE / CD138- labeled, and membrane-impermeable DNA dyes (DRAQ7) using flow cytometry analysis. FIG.6B shows target cell lysis of human CD3+ T cells activated with A-H.1 or OKT3 incubated with RPMI-8226 at a (E:T) ratio of 5:1 for 6 days followed by cell lysis analysis of RPMI 8226 cells as described above. Percentage (%) target cell lysis was determined by normalizing to basal target cell lysis (i.e., without antibody treatment) using the following formula, [(x - basal) / (100% - basal), where x is cell lysis of sample]. Data shown is a representative of n=1 donor.

[0164] FIGs. 7A-7B show IFNγ production by human PBMCs activated with the indicated antibodies.Human PBMCs were isolated from whole blood from the indicated number of donors, followed by solid- phase (plate-coated) stimulation with the indicated antibodies at 100Nm. Supernatant was collected on Days 1, 2, 3, 5, or 6. FIG.7A is a graph comparing the production of IFNγ in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) or anti-CD3e antibodies (OKT3 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG.7B shows IFNγ production in human PBMCs activated with the antibodies indicated activated with the indicated anti-TCR Vβ13.1 antibodies or anti-CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post-activation.

[0165] FIGs. 8A-8B show IL-2 production by human PBMCs activated with the indicated antibodies. Asimilar experimental setup as described for FIGs.7A-7B was used.

[0166] FIGs. 9A- 9B show IL-6 production by human PBMCs activated with the indicated antibodies.A similar experimental setup as described for FIGs.7A-7B was used.

[0167] FIGs. 10A- 10B show TNF-alpha production by human PBMCs activated with the indicatedantibodies. A similar experimental setup as described for FIGs.7A-7B was used.

[0168] FIGs. 11A- 11B show IL-1beta production by human PBMCs activated with the indicatedantibodies. A similar experimental setup as described for FIGs.7A-7B was used.

[0169] FIGs. 12A-12B are graphs showing delayed kinetics of IFNγ secretion in human PMBCsactivated by anti-TCR Vβ13.1 antibody A-H.1 when compared to PBMCs activated by anti-CD3e antibody OKT3. FIG.12A shows IFNγ secretion data from 4 donors. FIG.12B shows IFNγ secretion data from 4 additional donors. Data shown is representative of n=8 donors.WSGR Docket No.53676-768.601

[0170] FIG. 13 depicts increased CD8+ TSCM and Temra T cell subsets in human PBMCs activated byanti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) compared to PBMCs activated by anti-CD3e antibodies (OKT3 or SP34-2).

[0171] FIGs. 14A-14F show characterization of an anti-TCRVb antibody. FIG. 14A is a graphdepicting proliferation of T cells activated with anti-CD3 (OKT3) antibody or anti-TCRVb antibody. FIG.14B shows selective expansion of CD45RA+ effector memory CD8+ and CD4+ T cells (TEMRA) cells with anti- TCRVb antibodies. Tn= naïve T cell; Tscm= stem cell memory T cell; Tcm= central memory T cell; Tem=effector memory T cell; Temra=effector memory CD45RA+ T cell. FIG.14C is a graph showing IFN-g secretion by PBMCs stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies. FIG.14D shows target cell lysis by T cells stimulated with an anti-TCRVb antibody, or anti- CD3 antibodies. Cells were stimulated for 4 days followed by 2 days incubation with multiple myeloma target cells for assessment of cell killing. FIG.14E is a graph showing perforin secretion by T cells stimulated with an anti-TCRVb antibody, or an anti-CD3 antibody. Perforin was analyzed by FACS staining in TCRVB-positive and TCRVB-negative T cells in PBMCs after 5 days of stimulation with 100ng / ml plate-bound antibody. FIG.14F is a graph showing Granzyme B by T cells stimulated with an anti-TCRVb antibody, or an anti-CD3 antibody. Granzyme B was analyzed by FACS staining in TCRVB-positive and TCRVB-negative T cells in PBMCs after 5 days of stimulation with 100ng / ml plate-bound antibody.

[0172] FIGs. 15A-15B show production of IL-2 and IL-15 and expansion of human NK cells bystimulation of PBMCs with anti-TCRVb antibody for 6 days at a dose of 100nM. FIG.15A shows secretion of IL-2 or IL-15 in T cells stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies. FIG.15B depicts flow cytometry dot plots showing NKp46 staining vs CD56 antibody staining in cells stimulated with an anti-TCRVb antibody or an anti-CD3 antibody or a control sample.

[0173] FIGs. 16A-16C show secretion of cytokines in PBMCs stimulated with an anti-TCRVbantibody, or anti-CD3 antibodies.

[0174] FIGs. 17A-17B show killing of MM cells by dual targeting BCMA-TCRvb antibody molecules.FIG.17A shows in vitro killing by one of the following dual-targeting antibody molecules: BCMA- TCRVb (Molecule I), BCMA-CD3, or Control-TCRVb; or an isotype control. FIG.17B shows in vivo killing of MM cells by a dual-targeting BCM-TCRVb antibody (Molecule I).

[0175] FIG. 18 shows lysis of MM target cells with a dual targeting antibody (Molecule E) whichrecognized FcRH5 on one arm and TCRVb on the other arm.

[0176] FIGs. 19A-19B demonstrate cytokine production from human PBMCs activated by anti-TCRVβ8a antibodies (B-H.1) when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). FIG.19A shows that human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) produce similar or reduced levels of IFN^^. FIG.19B shows human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) produce higher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 6 donors.WSGR Docket No.53676-768.601

[0177] FIGs. 20A-20C demonstrate cytokine production from human PBMCs activated by anti-TCRVβ8a antibodies (B-H.1). Human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) do not significantly produce IL-6 (FIG.20A), IL1β (FIG.20B), and less TNFα (FIG.20C), when compared to PBMCs activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 6 donors.

[0178] FIGs. 21A-21E demonstrate cytokine production from human PBMCs activated by anti-TCRβVAntibody D antibody compared to control anti-CD3e antibody (OKT3). FIG.21A shows that human PBMCs activated by anti-TCRβV Antibody D antibody produce similar or reduced levels of IFN^^. FIG. 21B shows human PBMCs activated by anti-TCRβV Antibody D antibody produce higher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3). Human PBMCs activated by anti- TCRβV Antibody D antibody do not significantly produce IL-1beta (FIG.21C), IL-6, (FIG.21D), or TNFalpha (FIG.21E). Data shown is representative of n = 4 donors.

[0179] FIGs. 22A-22B demonstrate cytokine production from human PBMCs activated by anti-TCRVβ5 antibody (Antibody E). FIG.22A shows that human PBMCs activated by anti-TCR Vβ5 antibody produce similar or reduced levels of IFN^^ compared to PBMCS activated by anti-CD3^ antibodies (OKT3 or SP34-2). FIG.22B shows human PBMCs activated by the anti- TCR Vβ51 antibody producehigher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). Datashown is representative of n = 4 donors.

[0180] FIGs. 23A-23D demonstrate cytokine production from human PBMCs activated by an anti-TCRVβ5 antibody (Antibody E). Human PBMCs activated by anti-TCR Vβ5 antibody do not significantly produce IL-1beta (FIG.23A), IL-6, (FIG.23B), TNFalpha (FIG.23C), or IL-10 (FIG.23D) ascompared to PBMCs activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representativeof n = 4 donors.

[0181] FIGs. 24A-24F demonstrate cytokine production from human PBMCs activated by a dualtargeting (bispecific molecule) comprising an anti-TCRβV binding moiety and a BCMA binding moiety. FIG.24A shows that human PBMCs activated by the bispecific molecule produce similar or reduced levels of IFN^^ as PBMCS activated by anti-CD3^ antibodies (OKT3). FIG.24B shows human PBMCs activated by the bispecific molecule produce higher levels of IL-2 when compared to PBMCs activated by anti-CD3^ antibodies (OKT3). Human PBMCs activated by the bispecific molecule do not significantly produce IL-1beta (FIG.24C), IL-6, (FIG.24D), TNFalpha (FIG.24E), or IL-10 (FIG. 24F). Data shown is representative of n = 3 donors.

[0182] FIGs. 25A-25B show the structure and sequence of eight TCRβV proteins from seven differentsubfamilies: TCRβV6 subfamily (TCRβV6-5 and TCRβV6-4 are shown), TCRβV28 subfamily, TCRβV19 subfamily, TCRβV9 subfamily, TCRβV5 subfamily, TCRβV20 subfamily and TCRβV12 subfamily. FIG.25A shows the structural alignment of the different TCRβV proteins. The circled area represents the outward facing region comprising the proposed binding site for the anti-TCRβV antibodies as described herein. FIG.25B shows the amino acid sequence alignment of the proteins shown in FIG. 25A (SEQ ID NOS 3449-3456, respectively, in order of appearance). The various TCRβV proteins (fromWSGR Docket No.53676-768.601 7 different TCRβV subfamilies) have diverse sequences but share a conserved (similar) structure and function.

[0183] FIGs. 26A-26J show cytokine or chemokine secretion of PBMCs activated with anti-TCRVbantibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibody (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments. FIG.26A shows when plate-bound anti-TCR Vβ antibodies or anti-CD3e antibodies (OKT3) were used to activate human PBMCs, the T cell cytokine IFNg was induced. With respect to IL-2 production, PBMCs activated with anti-TCR Vβ antibodies resulted in increased IL-2 production with delayed kinetics (FIG.26B) as compared to PBMCs activated with anti- CD3e antibody (OKT3). While IL-1beta (FIG.26C), IL-6 (FIG.26D), IL-10 (FIG.26E), IL-4 (FIG. 26F), TNFalpha (FIG.26G), and IL-12p70 (FIG.26H) were induced by anti-CD3e antibody (OKT3), no or little induction of these cytokines or chemokines was observed with PBMCs activated with anti- TCRVb antibodies. PBMCs activated with anti-TCR Vβ antibodies demonstrated induction of IL-13 (FIG.26I) and IL-8 (FIG.26J).

[0184] FIGs. 27A-27H show cytokine or chemokine secretion of PBMCs activated with anti-TCRVbantibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibody (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments. PBMCs activated with anti-TCR Vβ antibodies demonstrated induction of Eotaxin (FIG.27A), Eotaxin 3 (FIG.27B), IL-8 or IL-18 (FIG.27C), MCP-1 (FIG.27E), MCP-4 (FIG.27F), MDC (FIG.27G), and MIP1a (FIG.27H). While IP-10 (FIG.27D) were induced by anti-CD3e antibody (OKT3), no or little induction of these cytokines or chemokines was observed with PBMCs activated with anti-TCRVb antibodies.

[0185] FIGs. 28A-28L show cytokine or chemokine secretion of PBMCs activated with anti-TCRVbantibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibody (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments. PBMCs activated with anti-TCR Vβ antibodies demonstrated induction of MIP1B (FIG.28A), TARC (FIG.28B), GM-CSF (FIG.28C), IL-15 (FIG. 28E), IL-16 (FIG.28F), and IL-15 (FIG.28I), and IL-7 (FIG.28J). While IL-12-23p40 (FIG.28D), IL- 17A (FIG.28G), and IL-1a (FIG.28H), were induced by anti-CD3e antibody (OKT3), no or little induction of these cytokines or chemokines was observed with PBMCs activated with anti-TCRVb antibodies.

[0186] FIG. 29 is a graph depicting mean tumor volume in NOD / SCID / IL-2Rγnull (NSG) miceengrafted with Raji-luc cells at days 10 to 28. The Star denotes PBMC implantation. Open triangles denote antibody treatment with the indicated antibodies.

[0187] FIGs. 30A-30F are graphs showing cytokine secretion stimulated by anti-TRBC1 (Antibody F)or anti-CD3 (OKT3) at Days 2 and 5. Cytokines examined include: IFNγ (FIG.30A), IL-2 (FIG.30B), IL-1β (FIG.30C), IL-6 (FIG.30D), IL-10 (FIG.30E), and TNFα (FIG.31F).WSGR Docket No.53676-768.601

[0188] FIG. 31 is a FACS plot showing the expansion of TCRvb 6-5+ T cells over 8 days using anti-TCRvb 6-5 v1.

[0189] FIG. 32 is a bar graph showing the expansion of TCRvb 6-5+ CD4+ T cells and TCRvb 6-5+CD8+ T cells over 8 days using the anti-CD3ε antibody OKT3 (100nM).

[0190] FIG. 33 is a bar graph showing the expansion of TCRvb 6-5+ CD4+ T cells and TCRvb 6-5+CD8+ T cells over 8 days using the anti-TCRvb 6-5 v1 antibody (100nM).

[0191] FIG. 34 is a FACS plot showing the showing the expansion of TCRvb 6-5+ T cells over 8 daysusing anti-TCRvb 6-5 v1 or the anti-CD3ε antibody OKT3.

[0192] FIG. 35A is a bar graph showing the percentage of TCRβV 6-5+ T cells in PBMC cultures after8 days of culture with the indicated antibody. Data for 5 replicates are shown. FIG.35B is a bar graph showing the percentage of TCRβV 6-5+ T cells in purified T cell cultures after 8 days of culture with the indicated antibody. Data for 5 replicates are shown.

[0193] FIG. 36A is a bar graph showing the relative count of TCRβV 6-5+ T cells in PBMC cultureafter 8 days of culture with the indicated antibody. FIG.36B is a bar graph showing the relative count of TCRβV 6-5+ T cells in PBMC culture after 8 days of culture with the indicated antibody.

[0194] FIG. 37A is a bar graph showing the relative count of TCRβV 6-5+ T cells in a purified T cellculture after 8 days of culture with the indicated antibody. FIG.37B is a bar graph showing the relative count of TCRβV 6-5+ T cells in a purified T cell culture after 8 days of culture with the indicated antibody.

[0195] FIG. 38 is a line graph showing the total CD3+ T cell count (fold increase) after 8 days of T cellculture with either the anti-CD3ε antibody OKT3 or the anti-TCRvb 6-5 v1 antibody.

[0196] FIG. 39 is a series of line graphs showing the kinetics of target cells by TCRβV 6-5 v1 activatedT cells or anti-CD3ε (OKT3) activated T cells. T cells from three different donors were utilized (donor 6769, donor 9880, donor 5411).

[0197] FIG. 40A is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1activated T cells or anti-CD3ε (OKT3) activated T cells without T cell pre activation. The data is presented at day 6 of co-culture between target cells and effector T cells. FIG.40B is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells with 4 days of T cell pre activation. The data is presented at day 2 of co-culture between target cells and effector T cells (after 4 days of T cell pre-activation).

[0198] FIG. 41 is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1activated T cells or anti-CD3ε (OKT3) activated T cells with 4 days of T cell pre activation. The data is presented at day 2 of co-culture between target cells and effector T cells (after 4 days of T cell pre- activation).

[0199] FIG. 42 is a bar graph showing target cell lysis by T cells by TCRβV 6-5 v1 activated T cells oranti-CD3ε (OKT3) activated T cells (100nM each antibody). The data includes seven replicates of each experimental condition.WSGR Docket No.53676-768.601

[0200] FIG. 43 is a series of FACS plots that show the cell surface expression of CD3ε on CD4+TCRβV 6-5- or CD4+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0201] FIG. 44 is a series of FACS plots that show the cell surface expression of CD3ε on CD8+TCRβV 6-5- or CD8+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0202] FIG. 45 is a series of FACS plots that show the cell surface expression of TCRβV on CD4+TCRβV 6-5- or CD4+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0203] FIG. 46 is a series of FACS plots that show the cell surface expression of TCRβV on CD8+TCRβV 6-5- or CD8+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0204] FIG. 47A shows FACS plot of TCRβV 6-5+ cynomolgus T cell expansion either unstimulated(left) or stimulated with anti-TCRβV 6-5 v1 (right) 7 days post activation of cynomolgus PBMCs. PBMCs from Donor DW8N (fresh PBMC sample, male, age 8, weight 7.9 kgs) were used. FIG.47B shows FACS plot of TCRβV 6-5+cynomolgus T cell expansion either unstimulated (left) or stimulated with anti-TCRβV 6-5 v1 (right) 7 days post activation of cynomolgus PBMCs. PBMCs from Donor G709 (cryopreserved sample, male, age 6, weight 4.7 kgs) were used.

[0205] FIG. 48 shows FACS plot and corresponding microscopy images of TCRβV 6-5+ cynomolgus Tcell expansion either unstimulated (left), stimulated with SP34-2 (anti-CD3ε antibody) (middle); or stimulated with anti-TCRβV 6-5 v1 (right) post activation of cryopreserved donor DW8N cynomolgus PBMCs. The microscopy images show the cell cluster formation (indicated by circles).

[0206] FIG. 49 shows a schematic of FACS plot showing the FACS gating / staining of PBMCs prior ɣ^T cell purification.

[0207] FIG. 50 shows a schematic of FACS plot showing the FACS gating / staining of purified ɣ^ T cellpopulation.

[0208] FIG. 51 show activation of purified ɣ^ T cell population with anti-CD3ε antibody (SP34-2) (left)or anti-TCRβV antibody (anti-TCRβV 6-5 v1) (right).

[0209] FIG. 52A shows the release of IFNɣ from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52B shows the release of TNFα from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52C shows the release of IL-2 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti- TCRβV 6-5 v1), or unstimulated. FIG.52D shows the release of IL-17A from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52E shows the release of IL-1α from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52F shows the release of IL-1β from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-WSGR Docket No.53676-768.601 2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52G shows the release of IL-6 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52H shows the release of IL-10 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated.

[0210] FIG. 53 shows the relative representations of all TCR alpha V segments (TRAV group of genes)and their variants (top), all TCR beta V segment 6-5 variants (TRBV6-5 gene) (bottom left), and all TCR beta V segments and variants excluding 6-5 (bottom right).

[0211] FIG. 54A is a FACS plot showing phenotypic markers of CD4+ T cells expanded with anti-TCRβV antibody (anti-TCRβV 6-5 v1). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right). FIG.54B is a FACS plot showing phenotypic markers of CD4+ T cells expanded with anti-CD3ε antibody (OKT3). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right).

[0212] FIG. 55A is a FACS plot showing phenotypic markers of CD8+ T cells expanded with anti-TCRβV antibody (anti-TCRβV 6-5 v1). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right). FIG.55B is a FACS plot showing phenotypic markers of CD8+ T cells expanded with anti-CD3ε antibody (OKT3). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right).

[0213] FIG. 56A is a bar graph showing the percentage of PD1 expressing CD4+ T cells from T cellcultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated. FIG.56B is a bar graph showing the percentage of PD1 expressing CD8+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0214] FIG. 57A is a bar graph showing the expression of Ki-67 by CD4+ T cells from T cell culturesactivated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated. FIG.57B is a bar graph showing the expression of Ki-67 by CD8+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0215] FIG. 58A is a FACS plot showing the percentage of TEMRA-like CD8+ T cells activated usinganti-TCRβV antibody (anti-TCRβV 6-5 v1) that express CD57 (18.7%). FIG.58B is a FACS plot showing the percentage of TEM-like CD8+ T cells activated using anti-CD3ε antibody (OKT3) that express CD57 (46.8%) and the percentage of TCM-like CD8+ T cells activated using anti-CD3ε antibody (OKT3) that express CD57 (18.9%).

[0216] FIG. 59 shows a series of FACS plots showing the expression of expression of CD27 and byCD4+ (top) or CD8+ (bottom) T cells from T cell cultures activated with anti-TCRβV antibody (anti- TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0217] FIG. 60 shows a series of FACS plots showing the expression of expression of OX40, 41BB,and ICOS by CD4+ (top) or CD8+ (bottom) T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.WSGR Docket No.53676-768.601

[0218] FIG. 61 shows a series of FACS plots showing the percentage of CD3+ (CD4 gated) TCRβV 6-5+ T cells 1, 2, 3, 4, 5, 6, and 8 days port activation with BCMA and the anti-TCR Vβ antibody anti-TCR Vβ 6-5 v1.

[0219] FIG. 62A shows a series of FACS plots showing the percentage of CD4+ T cells expanded usingisotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 0 post activation. FIG.62B shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 1 post activation. FIG.62C shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 2 post activation. FIG.62D shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti- TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 3 post activation. FIG.62E shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 4 post activation. FIG.62F shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 5 post activation. FIG.62G shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti- CD3ε (OKT3) antibodies on day 6 post activation. FIG.62H shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 8 post activation.

[0220] FIG. 63A is a bar graph showing ATP production from glycolysis of T cell cultures activatedwith the indicated antibodies. FIG.63B is a bar graph showing ATP production from oxidative phosphorylation of T cell cultures activated with the indicated antibodies.

[0221] FIG. 64 is a line graph showing the oxygen consumption rate (OCR) of T cells from about 0 to75 minutes activated with the indicated antibody.

[0222] FIG. 65A shows the oxygen consumption rate (OCR) of T cells activated with the indicatedantibody during basal respiration. FIG.65B shows the oxygen consumption rate (OCR) of T cells activated with the indicated antibody during maximal respiration. FIG.65C shows the oxygen consumption rate (OCR) of T cells activated with the indicated antibody during spare respiratory capacity. FIG.65D is a line graph indicates the areas of basal respiration and maximal respiration as shown in FIG.64A and FIG.64B, respectively.

[0223] FIG. 66A is a bar graph showing ATP production from glycolysis of T cell cultures activatedwith anti-TCRβV 6-5 v1 and re-stimulated with the indicated antibody. FIG.66B is a bar graph showing ATP production from oxidative phosphorylation of T cell cultures activated with anti-TCRβV 6-5 v1 and re-stimulated with the indicated antibody.

[0224] FIGs. 67A-67G are graphs showing expression of IFNγ (FIG. 67A), TNFα (FIG. 67E), IL-1α(FIG.67B), IL-1β (FIG.67C), IL-6 (CRS and neurotoxicity associated cytokines) (FIG.67D) withWSGR Docket No.53676-768.601 BHM1710 (anti TCRVB), a reduced affinity anti CD3 antibody (TB) and the SP34 anti CD3e antibody. IL-10 (FIG.67F), IL-17A (FIG.67G).

[0225] FIG. 68 is a FACS plot showing the percentage of NK cells expanded from T cell culturesactivated with the indicated antibody.

[0226] FIG. 69 is a bar graph showing the number of NK cells expanded from T cell cultures activatedwith the indicated antibody.

[0227] FIG. 70 shows a series of FACS plots showing NK cell proliferation induced by T cell culturesactivated with the indicated antibody.

[0228] FIG. 71 is a schematic showing an assay described in Example for determining NK cellmediated lysis of target K562 cells.

[0229] FIG. 72 is a bar graph showing the percent target cell lysis mediated by NK cells activated byPBMCs activated with the indicated antibody.

[0230] FIG. 73 shows a series of FACS plots showing the proliferation of NK cells from PBMCcultures activated / expanded with the indicated antibody (isotype control or OKT3). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0231] FIG. 74 shows a series of FACS plots showing the proliferation of NK cells from PBMCcultures activated / expanded with the indicated antibody (anti-TCRv^ 12-3 / 4 v1 or anti-TCRv^ 12-3 / 4 v2). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0232] FIG. 75 shows a series of FACS plots showing the proliferation of NK cells from PBMCcultures activated / expanded with the indicated antibody (anti-TCRv^ 12-3 / 4 v3 or SP34-2). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0233] FIG. 76 is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0234] FIG. 77 is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0235] FIG. 78 is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0236] FIG. 79 is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0237] FIG. 80 is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).WSGR Docket No.53676-768.601

[0238] FIG. 81 is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with theindicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0239] FIG. 82 is a bar graph showing the level of the indicated cytokine secreted by T cellsactivated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or SP34). The data includes use of 17 individual PBMC donors.

[0240] FIG. 83A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded withthe indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6).

[0241] FIG. 84A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded withthe indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti- TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicatedWSGR Docket No.53676-768.601 number of days (1, 2, 3, 5, or 6). FIG.84G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34- 2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6).

[0242] FIG. 85A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded withthe indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG. 85D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti- TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG. 85G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8).

[0243] FIG. 86A is a bar graph showing the level of secreted IL-17A by T cells activated / expanded withthe indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 7). FIG.86B is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 8). FIG.86C is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 7). FIG.86D is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or SP34-2) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7).

[0244] FIG. 87A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded withthe indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for theWSGR Docket No.53676-768.601 indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87C is a bar graph showing the level of secreted IL- 4 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87H is a bar graph showing the level of secreted IL-12p70 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87I is a bar graph showing the level of secreted IL- 13 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87J is a bar graph showing the level of secreted IL-8 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87K is a bar graph showing the level of secreted exotaxin by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87L is a bar graph showing the level of secreted exotoxin-3 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87M is a bar graph showing the level of secreted IL-8 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87N is a bar graph showing the level of secreted IP- 10 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with saidWSGR Docket No.53676-768.601 antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87O is a bar graph showing the level of secreted MCP-1 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87P is a bar graph showing the level of secreted MCP-4 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Q is a bar graph showing the level of secreted MDC by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87R is a bar graph showing the level of secreted MIP-1a by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87S is a bar graph showing the level of secreted MIP-1b by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87T is a bar graph showing the level of secreted TARC by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87U is a bar graph showing the level of secreted GMCSF by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87V is a bar graph showing the level of secreted IL-12-23p40 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87W is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87X is a bar graph showing the level of secreted IL- 16 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Y is a bar graph showing the level of secreted IL-17a by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Z is a bar graph showing the level of secreted IL-1a by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4WSGR Docket No.53676-768.601 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87AA is a bar graph showing the level of secreted IL-5 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87BB is a bar graph showing the level of secreted IL-7 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87CC is a bar graph showing the level of secreted TNF-B by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87DD is a bar graph showing the level of secreted VEGF by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8).

[0245] FIG. 88 shows a graphical representation of the relation of sequences between different TCRVBclonotype subfamilies.

[0246] FIG. 89A is a bar graph showing the percentage of cytokine release from PBMCsactivated / expanded for eight days using the indicated antibody (anti-TCRβV 12-3 / 4 v1 or SP34-2). FIG. 89B is a bar graph showing the percentage of cytokine release from PBMCs activated / expanded for eight days using the indicated antibody (anti-TCRβV 5 or SP34-2). FIG.89C is a bar graph showing the percentage of cytokine release from PBMCs activated / expanded for eight days using the indicated antibody (anti-TCRβV 10 or SP34-2).

[0247] FIG. 90A a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with theindicated antibody for the indicated number of days (3 or 6). FIG.90B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90G a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90H a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6).

[0248] FIG. 91 is a bar graph summarizing data from FACS analysis of PBMCs activated / expanded for6 days using the indicated anti-TCRV^ antibody.WSGR Docket No.53676-768.601

[0249] FIG. 92A a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with theindicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92G a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92H a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7).

[0250] FIG. 93 is a bar graph summarizing data from FACS analysis of PBMCs activated / expanded for7 days using the indicated anti-TCRV^ antibody.

[0251] FIG. 94A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded withthe indicated antibody for the indicated number of days (3 or 6). FIG.94B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94G a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94H a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94I a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6).

[0252] FIG. 95A is a bar graph showing the level of secreted IFN-ɣ by T cells activated / expanded withthe indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95B is a bar graph showing the level of secreted IFN-ɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6- 5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95C is a bar graph showing the level of secreted IL-1b by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody forWSGR Docket No.53676-768.601 the indicated number of days (1, 3, 5, or 7). FIG.95D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95F is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95G is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95H is a bar graph showing the level of secreted IL-1a by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95I is a bar graph showing the level of secreted IL-1b by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6- 5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95J is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95K is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95L is a bar graph showing the level of secreted TNF-a by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7).

[0253] FIG. 96 is a FACS plot showing the showing the ability of MH3-2 to bind PBMCs from one oftwo donors when the PBMCs are either preincubated with TM23 or not (MH3-2 Alone).

[0254] FIG. 97 is a FACS plot showing the ability of MH3-2 to bind PBMCs from one of two donorswhen the PBMCs are either preincubated with TM23 or not (MH3-2 Alone).

[0255] FIG. 98A is a bar graph showing the polyfunctional strength index (PSI) of PBMC CD4+ Tcells, CD4+ T cells expanded with anti-CD3 antibody, (CD3 Expanded T cells), and CD4+ T cells expanded with anti-TCRVβ 6-5 antibody (Drug Expanded T cells). The Effector mediators are Granzyme B, IFNɣ, MIP-1α, perforin, TNFα, and TNFβ. The Stimulatory mediators are IL-5. The Chemoattractive mediators are MIP-1b. FIG. 98B is a bar graph showing the polyfunctional strength index (PSI) ofWSGR Docket No.53676-768.601 PBMC CD8+ T cells, CD8+ T cells expanded with anti-CD3 antibody, (CD3 Expanded T cells), and CD8+ T cells expanded with anti-TCRVβ 6-5 antibody (Drug Expanded T cells). The Effector mediators are Granzyme B, IFNɣ, MIP-1α, perforin, and TNFβ. The Chemoattractive mediators are MIP-1b and RANTES.

[0256] FIG. 99 is a schematic of the experimental design for the pharmacokinetic (PK) profile anddosing strategy of the multifunctional polypeptide molecule as described herein.

[0257] FIG. 100 shows Table 9, which depicts alignment of TCRBV amino acid sequences (SEQ IDNOS 3457-3516, 3669-3673, 3522, 3674-3675, 3525, 3676-3687, 3538, 3688-3698, 3550-3639 and 3699-3790, respectively, in order of appearance). The alignment of TCRBV amino acid sequences in Table 9 underscores the diversity of TCR sequences. In particular, the TRBV sequences from different subfamilies are considerably different from each other.

[0258] FIG. 101 shows alignment of affinity matured humanized Antibody A-H VL sequences (SEQ IDNOS: 3377-3389, respectively, in order of appearance).

[0259] FIG. 102 shows alignment of affinity matured humanized Antibody A-H VH sequences (SEQ IDNOS: 3390-3436, respectively, in order of appearance).

[0260] FIG. 103A shows an exemplary embodiment (e.g., BKM0186) of the multifunctional moleculesas described herein comprising a first domain as described herein or a second domain as described herein and at least one cytokine or a functional fragment or variant thereof (e.g., IL2 or a functional fragment or variant thereof or IL2-C125A or a functional fragment or variant thereof) as described herein. FIG.103B shows an exemplary embodiment of the multifunctional molecules as described herein comprising a first domain as described herein or a second domain as described herein and at least one cytokine or a functional fragment or variant thereof as described herein. FIGs.103C, 103D, 103E, and 103F show exemplary embodiments of the multifunctional molecules as described herein comprising a first domain, a second domain, and two molecules of a cytokine or a functional fragment or variant thereof as described herein. In some embodiments, the cytokine or a functional fragment or variant thereof comprises IL-2 or a functional fragment or a functional variant thereof, IL2-C125A or a functional fragment or a functional variant thereof, IL-15 or a functional fragment or a functional variant thereof, IL-7 or a functional fragment or a functional variant thereof, IL-12 or a functional fragment or a functional variant thereof, or IL-21 or a functional fragment or a functional variant thereof. In embodiments, the cytokine polypeptide further comprises a cytokine receptor. In some embodiments, the cytokine polypeptide comprises IL-15 linked to a IL-15Ra. In some embodiments, the cytokine or a functional fragment or variant thereof as described herein comprises IL-15 or a functional fragment or variant thereof linked to a IL-15Ra sushi domain. In some embodiments, the cytokine or a functional fragment or variant thereof as described herein comprises a cytokine dimer. In some embodiments, the cytokine or a functional fragment or variant thereof as described herein comprises an IL-12 beta subunit or a functional fragment or variant thereof linked to an IL-12 alpha subunit or a functional fragment or variant thereof. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domainWSGR Docket No.53676-768.601 that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, the exemplary multifunctional molecules as described herein do not comprise at least one cytokine or a functional fragment or variant thereof.

[0261] FIG. 104 shows FACS plots showing binding of BKM0186 to different immune cell populationsin Human PBMCs.

[0262] FIG. 105 shows binding of BKM0186 to pure human T cells expressing either Vβ6 or CD25 (IL-2Rα) or both.

[0263] FIG. 106 shows in vitro concentration-effect relationships for BKM0186-mediated in vitroexpansion of Vβ6 T cells and activated (CD25) Vβ6 T cells from human PBMCs at day 5 as a % of total T-Cytotoxic (CD8) and T-helper (CD4) populations. Left graph: T-cytotoxic lymphocytes; right graph: T-helper lymphocytes.

[0264] FIG. 107 shows in vitro TCR sequencing. PBMCs were incubated with 100nM of BKM0186 for5 days and T cells were sequenced for TCR β chain V (TRBV) genes. Compared to unstimulated T cells (grey), BKM0186 selectively expanded T cells bearing TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-5, and TRBV10-3.

[0265] FIG. 108A and FIG. 108B show a series of graphs (FIG. 108A) and a series of FACS plots(FIG.108B) exhibiting activation of CD4+ and CD8+ T cells as assessed by CD25 expression following stimulation with BKM0186, RSV-IL2 and Anti-TCRVβ6 control in solution.

[0266] FIG. 109 shows a series of FACS plots demonstrating differentiation of memory T cellsmediated by BKM0186 in comparison to unstimulated and the controls RSV-IL2 and anti-TCRVβ6. Upper left quadrant represents Central memory (CM), lower left quadrant represents Effector memory (EM), upper right quadrant represents Naïve (N) and lower right quadrant represents Effector memory RA (TEMRA).

[0267] FIG. 110 shows in vitro concentration-effect relationships for BKM0186-induced cytokinerelease from human PBMCs at day 4 using MSD V-plex human cytokine panel.

[0268] FIG. 111 shows BKM0186-mediated killing of human tumor organoids generated from primary,patient-derived tissue from colorectal and NSCLC cancer patients. Vertical bars represent percentage of organoid area reduced relative to isotype control following incubation of organoids with BKM0186 and autologous TILs.

[0269] FIG. 112 shows tumor growth curves of mBKM0186-treated EMT6 tumor-bearing mice. Studieswere performed in randomized mice with tumor volumes of 80-150 mm3. For all models except MC38,WSGR Docket No.53676-768.601 mice were dosed for 3 weeks with a weekly dosing of 0.5-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point.

[0270] FIG. 113 shows tumor growth curves of mBKM0186-treated mice. Studies were performed inrandomized mice with tumor volumes of 80-150 mm3. For all models except MC38, mice were dosed for 4 weeks with a weekly dosing of 1-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point. For MC38, mice were given first dose of 3 mg / kg followed by 1 mg / kg for subsequent three weekly (QW) doses.

[0271] FIG. 114 shows Kaplan-Meier survival curves of treated mice. Studies were performed inrandomized mice with tumor volumes of 80-150 mm3. For all models except MC38, mice were dosed for 4 weeks with a weekly dosing of 1-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point. For MC38, mice were given first dose of 3 mg / kg followed by 1 mg / kg for subsequent three weekly (QW) doses.

[0272] FIG. 115 shows the experimental design for the tumor rechallenge study. Cured EMT6 tumorbearing mice were rechallenged with EMT6 tumor cells in one flank and CT26 tumor cells in another flank and monitored for tumor growth for 28 days.

[0273] FIG. 116 shows the results of the tumor rechallenge study. While the EMT6 tumors wererejected, CT26 tumors grew, suggesting that the memory response against EMT6 tumors likely mediated through mBKM0186 treatment has been established.

[0274] FIG. 117 shows immune profiling of T cells in blood and tumor tissue on day 14 post dosing ofmBKM0186.

[0275] FIG. 118 shows tumor growth curves of EMT6 tumors after weekly (QW) treatment of micebearing 150 mm3tumors with 1 mg / kg of mBKM0186 with and without depletion of Vβ-specific T cells. Filled Triangles indicate dosing intervals of the depleting antibodies and open Triangles indicate dosing intervals of mBKM0186.

[0276] FIG. 119A and FIG. 119B show Pharmacokinetic profiles of BKM0186 (FIG. 119A) andBKM0281 (FIG.119B) administered single dose IV in cynomolgus monkeys.

[0277] FIG. 120A shows T cell expansion following a single IV dose of BKM0186. FIG. 120B showsT cell expansion following a single IV dose BKM0281. n=3 Monkeys, n=1 monkey vehicle control.

[0278] FIG. 121 shows serum soluble CD25 levels in monkeys administered a single IV dose ofBKM0186. Mean values, n=2-3 monkeys per group.

[0279] FIG. 122A shows serum IL-6 levels in monkeys administered a single IV dose of BKM0186.FIG.122B shows serum IL-6 levels in monkeys administered a single IV dose of BKM0281. Mean values, n=2-3 monkeys per group.

[0280] FIG. 123A shows serum IFN-γ levels in monkeys administered a single IV dose of BKM0186.FIG.123B shows IFN-γ levels in monkeys administered a single IV dose of BKM0281. Mean values, n=2-3 monkeys per group. Mean values, n=2-3 monkeys per group.

[0281] FIG. 124 shows in vitro concentration-effect relationships for bispecific-mediated in vitroexpansion of Vβ6 T cells.WSGR Docket No.53676-768.601

[0282] FIGs. 125A and 125B show exemplary embodiments of the multifunctional molecules asdescribed herein comprising a first domain as described herein or a second domain as described herein and at least one cytokine or a functional fragment or variant thereof (e.g., wild-type human IL2 or a functional fragment or variant thereof or IL15-IL15R sushi fusion or a functional fragment or variant thereof) as described herein. FIGs.125C-125T show exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein. In some embodiments, a tumor-associated antigen (TAA)-binding moiety, a first domain as described herein and / or a second domain as described herein are antibody, an antigen binding fragment thereof or an antibody fragment. In some embodiments, an antigen binding fragment or an antibody fragment comprises Fab, Fab′, F(ab′)2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv). In some embodiments, the at least one cytokine or a functional fragment or variant thereof is selected from the group consisting of interleukin-2 (IL-2) or functional fragment or functional variant thereof, interleukin-7 (IL-7) or functional fragment or functional variant thereof, interleukin-12 (IL-12) or functional fragment or functional variant thereof, interleukin-15 (IL-15) or functional fragment or functional variant thereof, interleukin-18 (IL-18) or functional fragment or functional variant thereof, interleukin-21 (IL-21) or functional fragment or functional variant thereof, or interferon gamma or functional fragment or functional variant thereof, or any combination thereof. In some embodiments, exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein comprises a dimerization module comprising an Fc region comprising N297A mutation. FIGs.125C-125H show exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region comprising Knob-in-hole mutations and disulfide bridges. FIGs. 125I-125N show exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctionalWSGR Docket No.53676-768.601 molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region comprising disulfide bridges, but not comprising Knob-in-hole mutations. FIGs.125O-125T show exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region not comprising disulfide bridges nor Knob-in-hole mutations. FIGs.125C-125E, 125I-125K, and 125O-135Q show exemplary embodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, wherein a second domain as described herein or a first domain asdescribed herein comprises an scFv. FIGs. 125F-125H, 125L-125N, and 125R-135T show exemplaryembodiments of the multifunctional molecules as described herein, e.g., the multifunctional molecules comprising a first domain as described herein, a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, or the multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a first domain as described herein and / or a second domain as described herein, and at least one cytokine or a functional fragment or variant thereof as described herein, wherein a second domain as described herein or a first domain as described herein comprises an Fab. In some embodiments, exemplary embodiments of the multifunctional molecules as described herein comprises an antibody molecule, an antigen binding domain thereof, or a functional fragment or variant that binds to DLL-3 as an a-TAA. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, the exemplaryWSGR Docket No.53676-768.601 multifunctional molecules as described herein do not comprise at least one cytokine or a functional fragment or variant thereof.

[0283] FIG. 126 depicts exemplary graphs showing the average abundance of each depicted TRAVgene relative to total TRAV gene based on sequencing analysis of samples obtained from 5 healthy individuals. TCR repertoire sequencing was performed from total RNA isolated from healthy human purified T-cells. The MiXCR pipeline was used for alignment of reads to TCR clonotypes / germlines. Each TRAV gene is plotted against relative abundance (where 1 equals 100% of total TRAV gene). The average relative abundance for each TRAV is shown with each individual donor shown as (•). n=5.

[0284] FIG. 127A depicts exemplary flow cytometry results showing expansion of T-cells afterstimulation with an anti-TRAV 12-1 antibody. Purified human T-cells isolated from a healthy individual were stimulated for 5 days with plate-bound anti-TCR V alpha 12.1 antibody (6D6.6). The FACS plots show expanded T-cells stained with an AF647-labeled anti-TCR V alpha 12.1 antibody. Unstimulated T- cells were stained for baseline expression. Data from 1 representative donor is shown.

[0285] FIG. 127B depicts exemplary flow cytometry results showing expansion of T-cells afterstimulation with an anti-TRAV 12-1 antibody. Purified human T cells isolated from healthy individuals were stimulated for 10 days with 100 nM of plate-bound anti-TCR V alpha 12.1 antibody (6D6.6); followed by an additional 2-days in culture in the presence of 100 U / mL recombinant human IL-2. The FACS plots show expanded T cells stained with an AF647-labeled anti-TCR V alpha 12.1 antibody. Unstimulated T-cells were stained for baseline expression. Data from 2 representative donors are shown.

[0286] FIG. 128A depicts exemplary dot plots showing expansion of human CD4+ CD25+ and CD8+CD25+ T-cells after stimulation with anti-TCRαV-19 / IL-2 bispecific antibody. Human PBMCs were treated at with 0.001, 0.01, 0.1, 1, 10, or 100 nM of anti-TCRαV-19 / IL-2 for 5 days at 37°C. Cells were stained with anti-CD4, anti-CD25 (IL2RA), and anti-CD8 antibodies and FACS was used to quantify percentage of CD4+ CD25+ and CD8+ CD25+ T-cell populations. Isotype controls were used for baseline comparison.

[0287] FIG. 128B depicts exemplary dot plots showing expansion of murine CD4+ CD25+ and CD8+CD25+ T-cells after stimulation with anti-TCRαV-14 / IL-2 bispecific antibody. Isolated murine T-cells were treated at with 0.001, 0.01, 0.1, 1, 10, or 100 nM of anti-TCRαV-14 / IL-2 for 4 days at 37°C. Cells were stained with anti-CD4, anti-CD25 (IL2RA), and anti-CD8 antibodies and FACS was used to quantify percentage of CD4+ CD25+ and CD8+ CD25+ T-cell populations. Isotype controls were used for baseline comparison.

[0288] FIG. 128C depicts exemplary dot plots showing expansion of murine CD4+ CD25+ and CD8+CD25+ T-cells after stimulation with anti-TCRαV-12 / IL-2 bispecific antibody. Isolated murine T-cells were treated at with 0.001, 0.01, 0.1, 1, 10, or 100 nM of anti-TCRαV-12 / IL-2 for 4 days at 37°C. Cells were stained with anti-CD4, anti-CD25 (IL2RA), and anti-CD8 antibodies and FACS was used to quantify percentage of CD4+ CD25+ and CD8+ CD25+ T-cell populations. Isotype controls were used for baseline comparison.WSGR Docket No.53676-768.601

[0289] FIGs. 129A-129D depict exemplary embodiments of the multifunctional molecules as describedherein, wherein the N-terminus of the first portion of a dimerization module is linked to the C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the N-terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C- terminus of the first portion of the second domain. FIGs.129A and 129D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an scFv. FIGs.129B and 129C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an Fab. FIGs.129A and 129C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain as described herein comprises an scFv. FIGs.129B and 129D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain asdescribed herein comprises an Fab. In some embodiments, a first domain as described herein binds to a Tcell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain.

[0290] FIGs. 130A-130D depict exemplary embodiments of the multifunctional molecules as describedherein, wherein the N-terminus of the first portion of the dimerization module is linked to the C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the C-terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N- terminus of the first portion of the second domain. FIGs.130A and 130C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an Fab. FIGs.130B and 130D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an scFv. FIGs. 130A and 130D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain as described herein comprises an Fab. FIGs.130B and 130C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain asdescribed herein comprises an scFv. In some embodiments, a first domain as described herein binds to aT cell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, aWSGR Docket No.53676-768.601 second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain.

[0291] FIGs. 131A-131D depict exemplary embodiments of the multifunctional molecules as describedherein, wherein the C-terminus of the first portion of the dimerization module is linked to the N-terminus of the first domain or the N-terminus of the first portion of the first domain, and the C-terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N- terminus of the first portion of the second domain. FIGs.131A and 131D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an scFv. FIGs.131B and 131C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an Fab. FIGs.131A and 131B depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain as described herein comprises an scFv. FIGs.131C and 131D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain asdescribed herein comprises an Fab. In some embodiments, a first domain as described herein binds to a Tcell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain.

[0292] FIGs. 132A-132D depict exemplary embodiments of the multifunctional molecules as describedherein, wherein the C-terminus of the first portion of the dimerization module is linked to the N-terminus of the first domain or the N-terminus of the first portion of the first domain, and the N-terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C- terminus of the first portion of the second domain. FIGs.132A and 132B depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an Fab. FIGs.132C and 132D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a first domain as described herein comprises an scFv. FIGs. 132A and 132D depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain as described herein comprises an Fab. FIGs.132B and 132C depict exemplary embodiments of the multifunctional molecules as described herein, wherein a second domain asdescribed herein comprises an scFv. In some embodiments, a first domain as described herein binds to aT cell receptor alpha (TCRα) chain. In some embodiments, a second domain as described herein binds toWSGR Docket No.53676-768.601 a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor alpha (TCRα) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain. In some embodiments, a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. In some embodiments, a first domain as described herein binds to a T cell receptor beta (TCRβ) chain, and a second domain as described herein binds to a second domain that binds to a T cell receptor alpha (TCRα) chain. DETAILED DESCRIPTION DEFINITION

[0293] Certain specific details of this description are set forth in order to provide a thoroughunderstanding of various embodiments. However, one skilled in the art will understand that the present disclosure may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments.

[0294] Unless the context requires otherwise, throughout the specification and claims which follow, theword “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed disclosure.

[0295] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the”include plural referents unless the content clearly dictates otherwise. The use of the words “a” or “an” when used in conjunction with the term “comprising” herein may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0296] It should also be noted that the term “or” is generally employed in its sense including “and / or”unless the content clearly dictates otherwise.

[0297] Unless otherwise defined, all technical and scientific terms used herein have the same meaningas commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below.

[0298] The term “about” when referring to a measurable value such as an amount, a temporal duration,and the like, is meant to encompass variations of ±20% or in some instances ±10%, or in some instances ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. As used herein, “about” and “approximately” generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given range of values.

[0299] The term “acquire” or “acquiring” as the terms are used herein, refer to obtaining possession of aphysical entity (e.g., a sample, a polypeptide, a nucleic acid, or a sequence), or a value, e.g., a numericalWSGR Docket No.53676-768.601 value, by “directly acquiring” or “indirectly acquiring” the physical entity or value. “Directly acquiring” means performing a process (e.g., performing a synthetic or analytical method) to obtain the physical entity or value. “Indirectly acquiring” refers to receiving the physical entity or value from another party or source (e.g., a third party laboratory that directly acquired the physical entity or value). Directly acquiring a physical entity includes performing a process that includes a physical change in a physical substance, e.g., a starting material. Directly acquiring a value includes performing a process that includes a physical change in a sample or another substance, e.g., performing an analytical process which includes a physical change in a substance, e.g., a sample.

[0300] “Antibody molecule” as used herein refers to a protein, e.g., an immunoglobulin chain orfragment thereof, comprising at least one immunoglobulin variable domain structure and / or sequence. An antibody molecule encompasses antibodies (e.g., full-length antibodies) and antibody fragments. In some embodiments, an antibody molecule comprises an antigen binding or functional fragment of a full length antibody, or a full length immunoglobulin chain. For example, a full-length antibody is an immunoglobulin (Ig) molecule (e.g., an IgG antibody) that is naturally occurring or formed by normal immunoglobulin gene fragment recombinatorial processes). In embodiments, an antibody molecule refers to an immunologically active, antigen-binding portion of an immunoglobulin molecule, such as an antibody fragment. An antibody fragment, e.g., functional fragment, is a portion of an antibody, e.g., Fab, Fab′, F(ab′)2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv). A functional antibody fragment binds to the same antigen as that recognized by the intact (e.g., full-length) antibody. The terms “antibody fragment” or “functional fragment” also include isolated fragments consisting of the variable regions, such as the “Fv” fragments consisting of the variable regions of the heavy and light chains or recombinant single chain polypeptide molecules in which light and heavy variable regions are connected by a peptide linker (“scFv proteins”). In some embodiments, an antibody fragment does not include portions of antibodies without antigen binding activity, such as Fc fragments or single amino acid residues. Exemplary antibody molecules include full length antibodies and antibody fragments, e.g., dAb (domain antibody), single chain, Fab, Fab’, and F(ab’)2 fragments, a diabody, a VHH antibody, a nanobody, a single domain antibody, a single domain variant, a camelid antibody, and single chain variable fragments (scFvs). In some embodiments, the antibody molecule is an antibody mimetic. In some embodiments, the antibody molecule is, or comprises, an antibody-like framework or scaffold, such as, fibronectins, ankyrin repeats (e.g., designed ankyrin repeat proteins (DARPins)), avimers, affibody affinity ligands, anticalins, or affilin molecules.

[0301] The term “human-like antibody molecule” as used herein refers to a humanized antibodymolecule, human antibody molecule or an antibody molecule having at least 95% sequence identity with a non-murine germline framework region, e.g., FR1, FR2, FR3 and / or FR4. In some embodiments, the human-like antibody molecule comprises a framework region having at least 95% sequence identity to a human germline framework region, e.g., a FR1, FR2, FR3 and / or FR4 of a human germline framework region. In some embodiments, the human-like antibody molecule is a recombinant antibody. In some embodiments, the human-like antibody molecule is a humanized antibody molecule. In someWSGR Docket No.53676-768.601 embodiments, the human-like antibody molecule is human antibody molecule. In some embodiments, the human-like antibody molecule is a phage display or a yeast display antibody molecule. In some embodiments, the human-like antibody molecule is a chimeric antibody molecule. In some embodiments, the human-like antibody molecule is a CDR grafted antibody molecule.

[0302] As used herein, an “immunoglobulin variable domain sequence” refers to an amino acidsequence which can form the structure of an immunoglobulin variable domain. For example, the sequence may include all or part of the amino acid sequence of a naturally-occurring variable domain. For example, the sequence may or may not include one, two, or more N- or C-terminal amino acids, or may include other alterations that are compatible with formation of the protein structure.

[0303] In embodiments, an antibody molecule is monospecific, e.g., it comprises binding specificity fora single epitope. In some embodiments, an antibody molecule is multispecific, e.g., it comprises a plurality of immunoglobulin variable domain sequences, where a first immunoglobulin variable domain sequence has binding specificity for a first epitope and a second immunoglobulin variable domain sequence has binding specificity for a second epitope. In some embodiments, an antibody molecule is a bispecific antibody molecule. “Bispecific antibody molecule” as used herein refers to an antibody molecule that has specificity for more than one (e.g., two, three, four, or more) epitope and / or antigen.

[0304] “Antigen” (Ag) as used herein refers to a molecule that can provoke an immune response, e.g.,involving activation of certain immune cells and / or antibody generation. Any macromolecule, including almost all proteins or peptides, can be an antigen. Antigens can also be derived from genomic recombinant or DNA. For example, any DNA comprising a nucleotide sequence or a partial nucleotide sequence that encodes a protein capable of eliciting an immune response encodes an “antigen.” In embodiments, an antigen does not need to be encoded solely by a full length nucleotide sequence of a gene, nor does an antigen need to be encoded by a gene at all. In embodiments, an antigen can be synthesized or can be derived from a biological sample, e.g., a tissue sample, a tumor sample, a cell, or a fluid with other biological components. As used, herein a “tumor antigen” or interchangeably, a “cancer antigen” includes any molecule present on, or associated with, a cancer, e.g., a cancer cell or a tumor microenvironment that can provoke an immune response. As used, herein an “immune cell antigen” includes any molecule present on, or associated with, an immune cell that can provoke an immune response.

[0305] The “antigen-binding site,” or “binding portion” of an antibody molecule refers to the part of anantibody molecule, e.g., an immunoglobulin (Ig) molecule, that participates in antigen binding. In embodiments, the antigen binding site is formed by amino acid residues of the variable (V) regions of the heavy (H) and light (L) chains. Three highly divergent stretches within the variable regions of the heavy and light chains, referred to as hypervariable regions, are disposed between more conserved flanking stretches called “framework regions,” (FRs). FRs are amino acid sequences that are naturally found between, and adjacent to, hypervariable regions in immunoglobulins. In embodiments, in an antibody molecule, the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relative to each other in three dimensional space to form an antigen-binding surface,WSGR Docket No.53676-768.601 which is complementary to the three-dimensional surface of a bound antigen. The three hypervariable regions of each of the heavy and light chains are referred to as “complementarity-determining regions,” or “CDRs.” The framework region and CDRs have been defined and described, e.g., in Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.91-3242, and Chothia, C. et al. (1987) J. Mol. Biol.196:901-917 or by ImMunoGeneTics (IMGT) system. Each variable chain (e.g., heavy chain variable region and light chain variable region) is typically made up of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the amino acid order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0306] As used herein, an “immune cell” refers to any of various cells that function in the immunesystem, e.g., to protect against agents of infection and foreign matter. In embodiments, this term includes leukocytes, e.g., neutrophils, eosinophils, basophils, lymphocytes, and monocytes. Innate leukocytes include phagocytes (e.g., macrophages, neutrophils, and dendritic cells), mast cells, eosinophils, basophils, and natural killer cells. Innate leukocytes identify and eliminate pathogens, either by attacking larger pathogens through contact or by engulfing and then killing microorganisms, and are mediators in the activation of an adaptive immune response. The cells of the adaptive immune system are special types of leukocytes, called lymphocytes. B cells and T cells are important types of lymphocytes and are derived from hematopoietic stem cells in the bone marrow. B cells are involved in the humoral immune response, whereas T cells are involved in cell-mediated immune response. The term “immune cell” includes immune effector cells.

[0307] “Immune effector cell,” as that term is used herein, refers to a cell that is involved in an immuneresponse, e.g., in the promotion of an immune effector response. Examples of immune effector cells include, but are not limited to, T cells, e.g., alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NK T) cells, and mast cells.

[0308] The term “effector function” or “effector response” refers to a specialized function of a cell.Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines.

[0309] The terms “polypeptide”, “peptide” and “protein” (if single chain) are used interchangeablyherein to refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component. The polypeptide can be isolated from natural sources, can be a produced by recombinant techniques from a eukaryotic or prokaryotic host, or can be a product of synthetic procedures.

[0310] The terms “nucleic acid,” “nucleic acid sequence,” “nucleotide sequence,” or “polynucleotidesequence,” and “polynucleotide” are used interchangeably. They refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. The polynucleotide may be either single-stranded or double-stranded, and if single-stranded may be the coding strand or non- coding (antisense) strand. A polynucleotide may comprise modified nucleotides, such as methylatedWSGR Docket No.53676-768.601 nucleotides and nucleotide analogs. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component. The nucleic acid may be a recombinant polynucleotide, or a polynucleotide of genomic, cDNA, semisynthetic, or synthetic origin which either does not occur in nature or is linked to another polynucleotide in a non-natural arrangement.

[0311] The term “isolated,” as used herein, refers to material that is removed from its original or nativeenvironment (e.g., the natural environment if it is naturally occurring). For example, a naturally- occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide, separated by human intervention from some or all of the co-existing materials in the natural system, is isolated. Such polynucleotides could be part of a vector and / or such polynucleotides or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of the environment in which it is found in nature. An isolated polynucleotide (ribonucleic acid (RNA), deoxyribonucleic acid (DNA)), or polypeptide is free of the genes / nucleic acids or sequences / amino acids that flank it in its naturally-occurring state.

[0312] The compositions and methods of the present invention encompass polypeptides and nucleicacids having the sequences specified, or sequences substantially identical or similar thereto, e.g., sequences at least 80%, 85%, 90%, 95% identical or higher to the sequence specified. In the context of an amino acid sequence, the term “substantially identical” is used herein to refer to a first amino acid that contains a sufficient or minimum number of amino acid residues that are i) identical to, or ii) conservative substitutions of aligned amino acid residues in a second amino acid sequence such that the first and second amino acid sequences can have a common structural domain and / or common functional activity. For example, amino acid sequences that contain a common structural domain having at least about 80%, 85%, 90%.91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99%, 99.5%, 99.9%, or 100% sequence identity to a reference sequence, e.g., a sequence provided herein. In the context of nucleotide sequence, the term “substantially identical” is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity, or encode a common structural polypeptide domain or a common functional polypeptide activity. For example, nucleotide sequences having at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99%, 99.5%, 99.9%, or 100% sequence identity to a reference sequence, e.g., a sequence provided herein.

[0313] The term “variant” refers to a polypeptide that has a substantially identical amino acid sequenceto a reference amino acid sequence, or is encoded by a substantially identical nucleotide sequence. In some embodiments, the variant is a functional variant. In some embodiments, a TCRβV variant can bind to TCRα and form a TCR α:β complex.

[0314] The term “functional variant” refers to a polypeptide that has a substantially identical amino acidsequence to a reference amino acid sequence, or is encoded by a substantially identical nucleotide sequence, and is capable of having one or more activities of the reference amino acid sequence.WSGR Docket No.53676-768.601

[0315] The term “functional fragment” refers to a polypeptide that has a partial amino acid sequence ofa reference amino acid sequence, and is capable of having one or more activities of the reference amino acid sequence. In some embodiments, the functional fragment comprises at least about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99%, 99.5%, or 99.9% amino acid sequence of a reference amino acid sequence. In some embodiments, the functional fragment comprises an amino acid sequence that has at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acid deletion from the reference amino acid sequence. In some embodiments, the functional fragment comprises an amino acid sequence that has at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 350, 400, 450, or 500 amino acids of the reference amino acid sequence.

[0316] Calculations of homology or sequence identity between sequences (the terms are usedinterchangeably herein) are performed as follows. To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”).

[0317] The percent identity between the two sequences is a function of the number of identical positionsshared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol.48:444-453 ) algorithm which has been incorporated into the GAP program in the GCG software package (available at www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and the one that should beWSGR Docket No.53676-768.601 used unless otherwise specified) are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0318] The percent identity between two amino acid or nucleotide sequences can be determined usingthe algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4:11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The nucleic acid and protein sequences described herein can be used as a “query sequence” to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol.215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to a nucleic acid molecule of the invention. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res.25:3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0319] It is understood that the molecules of the present invention may have additional conservative ornon-essential amino acid substitutions, which do not have a substantial effect on their functions.

[0320] The term “amino acid” is intended to embrace all molecules, whether natural or synthetic, whichinclude both an amino functionality and an acid functionality and capable of being included in a polymer of naturally-occurring amino acids. Exemplary amino acids include naturally-occurring amino acids; analogs, derivatives and congeners thereof; amino acid analogs having variant side chains; and all stereoisomers of any of any of the foregoing. As used herein the term “amino acid” includes both the D- or L- optical isomers and peptidomimetics.

[0321] A “conservative amino acid substitution” is one in which the amino acid residue is replaced withan amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0322] As used herein, the term “molecule” as used in, e.g., antibody molecule, cytokine molecule,receptor molecule, includes full-length, naturally-occurring molecules, as well as variants, e.g., functional variants (e.g., truncations, fragments, mutated (e.g., substantially similar sequences) or derivatized form thereof), so long as at least one function and / or activity of the unmodified (e.g., naturally-occurring) molecule remains.WSGR Docket No.53676-768.601

[0323] As used herein, the term “mutation” refers to an alteration in the nucleotide sequence of thegenome of an organism, virus, or extrachromosomal DNA. In some embodiments, the mutation may be a large-scale mutation, such as amplifications (or gene duplications) or repetitions of a chromosomal segment, deletions of large chromosomal regions, chromosomal rearrangements (e.g., chromosomal translocations, chromosomal inversions, non-homologous chromosomal crossover, and interstitial deletions), and loss of heterozygosity. In some embodiments, the mutation may be a small-scale mutation, such as insertions, deletions, and substitution mutations. As used herein, the term “substitution mutation” refers to the transition that exchange a single nucleotide for another.

[0324] As used herein, the term “linked” and “operatively linked” are interchangeably used.

[0325] “Interleukin-2” also known as IL2, IL-2, IL 2, TCGF, lymphokine, and interleukin 2, as referredto herein, includes any of the recombinant or naturally-occurring forms of IL-2 or variants or homologs thereof that have or maintain IL-2 activity (e.g., at least 40% 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity). In some aspects, the variants or homologs have at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring IL-2. In some embodiments, IL-2 is substantially identical to the protein identified by the UniProt reference number P60568 or a variant or homolog having substantial identity thereto. Human T cell receptor (TCR) complex

[0326] TCR is a disulfide-linked membrane-anchored heterodimeric protein normally consisting of thehighly variable alpha (α) and beta (β) chains expressed as part of a complex with the invariant CD3 chain molecules. TCR on αβ T cells is formed by a heterodimer of one alpha chain and one beta chain. Each alpha or beta chain consists of a constant domain and a highly variable domain classified as the Immunoglobulin superfamily (IgSF) fold. The TCRβV chains can be further classified into subfamilies. Despite their high structural and functional homology, the amino acid sequence homology in the TRBV genes is very low. Only 4 amino acids out of approximately 95 are identical while 10 additional amino acids are conserved among all subfamilies. Nevertheless, TCRs formed between alpha and beta chains of highly diverse sequences show a remarkable structural homology and elicit a similar function, e.g., activation of T cells.

[0327] T cell receptors (TCR) can be found on the surface of T cells. TCRs recognize antigens, e.g.,peptides, presented on, e.g., bound to, major histocompatibility complex (MHC) molecules on the surface of cells, e.g., antigen-presenting cells. TCRs are heterodimeric molecules and can comprise an alpha chain, a beta chain, a gamma chain or a delta chain. TCRs comprising an alpha chain and a beta chain are also referred to as TCRαβ. The TCR beta chain consists of the following regions (also known as segments): variable (V), diversity (D), joining (J) and constant (C) (see Mayer G. and Nyland J. (2010) Chapter 10: Major Histocompatibility Complex and T-cell Receptors-Role in Immune Responses. In: Microbiology and Immunology on-line, University of South Carolina School of Medicine). The TCRWSGR Docket No.53676-768.601 alpha chain consists of V, J and C regions. The rearrangement of the T-cell receptor (TCR) through somatic recombination of V (variable), D (diversity), J (joining), and C (constant) regions is a defining event in the development and maturation of a T cell. TCR gene rearrangement takes place in the thymus.

[0328] TCRs can comprise a receptor complex, known as the TCR complex, which comprises a TCRheterodimer comprising of an alpha chain and a beta chain, and dimeric signaling molecules, e.g., CD3 co-receptors, e.g., CD3δ / ε, and / or CD3γ / ε.

[0329] As used herein, the term “T cell receptor beta variable chain” or “TCRβV,” refers to anextracellular region of the T cell receptor beta chain which comprises the antigen recognition domain of the T cell receptor. The term TCRβV includes isoforms, mammalian, e.g., human TCRβV, species homologs of human and analogs comprising at least one common epitope with TCRβV. Human TCRβV comprises a gene family comprising subfamilies including, but not limited to: a TCRβ V6 subfamily, a TCRβ V10 subfamily, a TCRβ V12 subfamily, a TCRβ V5 subfamily, a TCRβ V7 subfamily, a TCRβ V11 subfamily, a TCRβ V14 subfamily, a TCRβ V16 subfamily, a TCRβ V18 subfamily, a TCRβ V9 subfamily, a TCRβ V13 subfamily, a TCRβ V4 subfamily, a TCRβ V3 subfamily, a TCRβ V2 subfamily, a TCRβ V15 subfamily, a TCRβ V30 subfamily, a TCRβ V19 subfamily, a TCRβ V27 subfamily, a TCRβ V28 subfamily, a TCRβ V24 subfamily, a TCRβ V20 subfamily, TCRβ V25 subfamily, a TCRβ V29 subfamily, a TCRβ V1 subfamily, a TCRβ V17 subfamily, a TCRβ V21 subfamily, a TCRβ V23 subfamily, or a TCRβ V26 subfamily, as well as family members of said subfamilies, and variants thereof (e.g., a structural or functional variant thereof). In some embodiments, the TCRβ V6 subfamily comprises: TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-9*01, TCRβ V6-8*01, TCRβ V6-5*01, TCRβ V6-6*02, TCRβ V6-6*01, TCRβ V6-2*01, TCRβ V6-3*01 or TCRβ V6-1*01. In some embodiments, TCRβV comprises TCRβ V6-5*01, or a variant thereof, e.g., a variant having 85%, 90%, 95%, 99% or more identity the naturally-occurring sequence. TCRβ V6-5*01 is also known as TRBV65; TCRBV6S5; TCRBV13S1, or TCRβ V13.1. The amino acid sequence of TCRβ V6-5*01, e.g., human TCRβ V6-5*01, is known in that art, e.g., as provided by IMGT ID L36092. In some embodiments, TCRβ V6-5*01 is encoded by the nucleic acid sequence of SEQ ID NO: 43, or a sequence having 85%, 90%, 95%, 99% or more identity thereof. In some embodiments, TCRβ V6-5*01 comprises the amino acid sequence of SEQ ID NO: 44, or a sequence having 85%, 90%, 95%, 99% or more identity thereof.

[0330] SEQ ID NO: 43ATGAGCATCGGCCTCCTGTGCTGTGCAGCCTTGTCTCTCCTGTGGGCAGGTCCAGTGAATGC TGGTGTCACTCAGACCCCAAAATTCCAGGTCCTGAAGACAGGACAGAGCATGACACTGCAG TGTGCCCAGGATATGAACCATGAATACATGTCCTGGTATCGACAAGACCCAGGCATGGGGC TGAGGCTGATTCATTACTCAGTTGGTGCTGGTATCACTGACCAAGGAGAAGTCCCCAATGGC TACAATGTCTCCAGATCAACCACAGAGGATTTCCCGCTCAGGCTGCTGTCGGCTGCTCCCTC CCAGACATCTGTGTACTTCTGTGCCAGCAGTTACTC

[0331] SEQ ID NO: 44MSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMG LRLIHY-SVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYWSGR Docket No.53676-768.601 TCR beta V (TCRβV)

[0332] Diversity in the immune system enables protection against a huge array of pathogens. Since thegermline genome is limited in size, diversity is achieved not only by the process of V(D)J recombination but also by junctional (junctions between V-D and D-J segments) deletion of nucleotides and addition of pseudo-random, non-templated nucleotides. The TCR beta gene undergoes gene arrangement to generate diversity.

[0333] The TCR V beta repertoire varies between individuals and populations because of, e.g., 7frequently occurring inactivating polymorphisms in functional gene segments and a large insertion / deletion-related polymorphism encompassing 2 V beta gene segments.

[0334] Provided herein are, inter alia, antibody molecules and fragments thereof, that bind, e.g.,specifically bind, to a human TCR beta V chain (TCRβV), e.g., a TCRβV gene family (also referred to as a group), e.g., a TCRβV subfamily (also referred to as a subgroup), e.g., as described herein. TCR beta V families and subfamilies are known in the art, e.g., as described in Yassai et al., (2009) Immunogenetics61(7)pp:493-502; Wei S. and Concannon P. (1994) Human Immunology 41(3) pp: 201-206. Theantibodies described herein can be recombinant antibodies, e.g., recombinant non-murine antibodies, e.g., recombinant human or humanized antibodies.

[0335] The terms TCRBV, TCRVB, TRBV, TCRβV, TCRVβ or TRβV are used interchangeably hereinand refer to a TCR beta V chain, e.g., as described herein.

[0336] The terms TCRBV, TCRVB, TRBV, TCRβV, TCRVβ or TRβV are used interchangeably hereinand refer to a TCR beta V chain, e.g., as described herein.

[0337] In some embodiments, provided herein is an anti-TCRβV antibody molecule that binds to humanTCRβV, e.g., a TCRβV family, e.g., gene family or a variant thereof. In some embodiments a TCRBV gene family comprises one or more subfamilies, e.g., as described herein, e.g., Table 8A or Table 8B. In some embodiments, the TCRβV gene family comprises: a TCRβ V6 subfamily, a TCRβ V10 subfamily, a TCRβ V12 subfamily, a TCRβ V5 subfamily, a TCRβ V7 subfamily, a TCRβ V11 subfamily, a TCRβ V14 subfamily, a TCRβ V16 subfamily, a TCRβ V18 subfamily, a TCRβ V9 subfamily, a TCRβ V13 subfamily, a TCRβ V4 subfamily, a TCRβ V3 subfamily, a TCRβ V2 subfamily, a TCRβ V15 subfamily, a TCRβ V30 subfamily, a TCRβ V19 subfamily, a TCRβ V27 subfamily, a TCRβ V28 subfamily, a TCRβ V24 subfamily, a TCRβ V20 subfamily, TCRβ V25 subfamily, a TCRβ V29 subfamily, a TCRβ V1 subfamily, a TCRβ V17 subfamily, a TCRβ V21 subfamily, a TCRβ V23 subfamily, or a TCRβ V26 subfamily.

[0338] In some embodiments, TCRβ V6 subfamily is also known as TCRβ V13.1. In someembodiments, the TCRβ V6 subfamily comprises: TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-9*01, TCRβ V6-8*01, TCRβ V6-5*01, TCRβ V6-6*02, TCRβ V6-6*01, TCRβ V6-2*01, TCRβ V6-3*01 or TCRβ V6-1*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-9*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-8*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβWSGR Docket No.53676-768.601 V6-5*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-2*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-3*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-1*01, or a variant thereof.

[0339] In some embodiments, TCRβ V6 comprises TCRβ V6-5*01, or a variant thereof. In someembodiments, TCRβ V6, e.g., TCRβ V6-5*01, is recognized, e.g., bound, by SEQ ID NO: 1 and / or SEQ ID NO: 2. In some embodiments, TCRβ V6, e.g., TCRβ V6-5*01, is recognized, e.g., bound, by SEQ ID NO: 9 and / or SEQ ID NO: 10. In some embodiments, TCRβ V6 is recognized, e.g., bound, by SEQ ID NO: 9 and / or SEQ ID NO: 11.

[0340] In some embodiments, TCRβ V10 subfamily is also known as TCRβ V12. In someembodiments, the TCRβ V10 subfamily comprises: TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-3*01 or TCRβ V10-2*01, or a variant thereof.

[0341] In some embodiments, TCRβ V12 subfamily is also known as TCRβ V8.1. In someembodiments, the TCRβ V12 subfamily comprises: TCRβ V12-4*01, TCRβ V12-3*01, or TCRβ V12- 5*01, or a variant thereof. In some embodiments, TCRβ V12 is recognized, e.g., bound, by SEQ ID NO: 15 and / or SEQ ID NO: 16. In some embodiments, TCRβ V12 is recognized, e.g., bound, by any one of SEQ ID NOs 23-25, and / or any one of SEQ ID NO: 26-30:

[0342] In some embodiments, the TCRβ V5 subfamily is chosen from: TCRβ V5-5*01, TCRβ V5-6*01,TCRβ V5-4*01, TCRβ V5-8*01, TCRβ V5-1*01, or a variant thereof.

[0343] In some embodiments, the TCRβ V7 subfamily comprises TCRβ V7-7*01, TCRβ V7-6*01,TCRβ V7 -8*02, TCRβ V7 -4*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*01, TCRβ V7-3*01, TCRβ V7-9*03, or TCRβ V7-9*01, or a variant thereof.

[0344] In some embodiments, the TCRβ V11 subfamily comprises: TCRβ V11-1*01, TCRβ V11-2*01or TCRβ V11-3*01, or a variant thereof. In some embodiments, the TCRβ V14 subfamily comprises TCRβ V14*01, or a variant thereof. In some embodiments, the TCRβ V16 subfamily comprises TCRβ V16*01, or a variant thereof. In some embodiments, the TCRβ V18 subfamily comprises TCRβ V18*01, or a variant thereof. In some embodiments, the TCRβ V9 subfamily comprises TCRβ V9*01 or TCRβ V9*02, or a variant thereof. In some embodiments, the TCRβ V13 subfamily comprises TCRβ V13*01, or a variant thereof. In some embodiments, the TCRβ V4 subfamily comprises TCRβ V4-2*01, TCRβ V4-3*01, or TCRβ V4-1*01, or a variant thereof. In some embodiments, the TCRβ V3 subfamily comprises TCRβ V3-1*01, or a variant thereof. In some embodiments, the TCRβ V2 subfamily comprises TCRβ V2*01, or a variant thereof. In some embodiments, the TCRβ V15 subfamily comprises TCRβ V15*01, or a variant thereof. In some embodiments, the TCRβ V30 subfamily comprises TCRβ V30*01, or TCRβ V30*02, or a variant thereof. In some embodiments, the TCRβ V19 subfamily comprises TCRβ V19*01, or TCRβ V19*02, or a variant thereof. In some embodiments, the TCRβ V27 subfamily comprises TCRβ V27*01, or a variant thereof. In some embodiments, the TCRβ V28 subfamily comprises TCRβ V28*01, or a variant thereof. In some embodiments, the TCRβ V24WSGR Docket No.53676-768.601 subfamily comprises TCRβ V24-1*01, or a variant thereof. In some embodiments, the TCRβ V20 subfamily comprises TCRβ V20-1*01, or TCRβ V20-1*02, or a variant thereof. In some embodiments, the TCRβ V25 subfamily comprises TCRβ V25-1*01, or a variant thereof. In some embodiments, the TCRβ V29 subfamily comprises TCRβ V29-1*01, or a variant thereof.

[0345] Exemplary amino acid sequences for TCRβV subfamily members can be found on theImMunoGeneTics Information System website: www.imgt.org / , or in a similar resource. Anti-TCRβV antibodies

[0346] Current bispecific constructs designed to redirect T cells to promote tumor cell lysis for cancerimmunotherapy typically utilize antibody fragments (Fab, scFv, VH, single domain antibody, etc.) that are derived from monoclonal antibodies (mAb) directed against the CD3e subunit of the T cell receptor (TCR). However, there are limitations to this approach which may prevent the full realization of the therapeutic potential for such bispecific constructs. Previous studies have shown that even low “activating” doses of anti-CD3e mAb can cause long-term T cell dysfunction and exert immunosuppressive effects. In addition, anti-CD3e mAbs have been associated with side effects that result from massive T cell activation. The large number of activated T cells secrete substantial amounts of cytokines, the most important of which is Interferon gamma (IFNγ). This excess amount of IFNγ in turn activates macrophages which then overproduce proinflammatory cytokines such as IL-1beta, IL-6, IL-10 and TNF-alpha, causing a “cytokine storm” known as the cytokine release syndrome (CRS) (Shimabukuro-Vornhagen et al., J Immunother Cancer.2018 Jun 15;6(1):56, herein incorporated by reference in its entirety). Thus, the need exists for developing antibodies that are capable of binding and activating only a subset of effector T cells, e.g., to re-duce the CRS and / or neurotoxicity (NT).

[0347] Described herein are molecules targeting the TCRβV chain of TCR and methods thereof.Without wishing to be bound by theory, such molecules are capable of binding, activating, and / or expanding only a subset of T cells, avoiding or reducing CRS and / or NT and minimizing potential immunosuppressive effects of anti-CD3 mAbs.

[0348] Described herein is a class of antibodies, i.e., anti-TCRβV antibody molecules as describedherein, which despite having low sequence similarity (e.g., low sequence identity among the different antibody molecules that recognize different TCRβV subfamilies), recognize a structurally conserved, yet sequence-wise variable, region, e.g., domain, on the TCRβV protein and have a similar function (e.g., activation of T cells and a similar cytokine profile as described herein). Thus, the anti-TCRβV antibody molecules as described herein share a structure-function relationship.

[0349] Without wishing to be bound by theory, in some embodiments, the anti-TCRβV antibodymolecules as described herein bind to an outward facing epitope of a TCRβV protein when it is in a complex with a TCRalpha protein. In some embodiments, the anti-TCRβV antibody molecules as described herein recognize (e.g., bind to), a domain (e.g., an epitope) on the TCRβV protein that is: (1) structurally conserved among different TCRβV subfamilies; and (2) has minimal sequence identity among the different TCRβV subfamilies. TCRβV proteins from the different TCRBV subfamilies shareWSGR Docket No.53676-768.601 minimal sequence similarity. However, TCRβV proteins which have minimal sequence similarity, share a similar 3D conformation and structure.

[0350] The alignment of TCRBV amino acid sequences in ta9 underscores the diversity of TCRsequences. In particular, the TRBV sequences from different subfamilies are considerably different from each other.

[0351] In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize,e.g., bind to, an interface of a TCRβV:TCRalpha complex. In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize, e.g., bind to, a constant region of a TCRβV protein. An exemplary antibody that binds to a constant region of a TCRBV region is JOVI.1 as de- scribed in Viney et al., (Hybridoma.1992 Dec;11(6):701-13). In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize, e.g., bind to, one or more (e.g., all) of a complementarity determining region (e.g., CDR1, CDR2 and / or CDR3) of a TCRβV protein.

[0352] Provided herein are, inter alia, antibody molecules directed to the variable chain of the betasubunit of TCR (TCRβV) which bind and, e.g., activate a subset of T cells. The anti-TCRβV antibody molecules as described herein result in lesser or no production of cytokines associated with CRS, e.g., IL- 6, IL-1beta, IL-10 and TNF alpha; and enhanced and / or delayed production of IL-2 and IFNγ. In some embodiments, the anti-TCRβV antibodies as described herein have a cytokine profile, e.g., as described herein, which differs from a cytokine profile of a T cell engager that binds to a receptor or molecule other than a TCRβV region (“a non-TCRβV-binding T cell engager”). In some embodiments, the non-TCRβV- binding T cell engager comprises an antibody that binds to a CD3 molecule (e.g., CD3 epsilon (CD3e) molecule); or a TCR alpha (TCRα) molecule. In some embodiments, the non-TCRβV-binding T cell engager is an OKT3 antibody or an SP34-2 antibody.

[0353] In some embodiments, the anti-TCRβV antibodies as described herein result in expansion ofTCRβV+ T cells, e.g., a subset of memory effector T cells known as TEMRA. Without wishing to be bound by theory, it is believed that in some embodiments, TEMRA cells can promote tumor cell lysis but not CRS. Accordingly, provided herein are methods of making said anti-TCRβV antibody molecules and uses thereof. Also described herein are multispecific molecules, e.g., bispecific molecules comprising said anti-TCRβV antibody molecules. In some embodiments, compositions comprising anti-TCRβV antibody molecules of the present disclosure, can be used, e.g., to: (1) activate and redirect T cells to promote tumor cell lysis for cancer immuno-therapy; and / or (2) expand TCRβV+ T cells. In some embodiments, compositions comprising anti-TCRβV antibody molecules as described herein limit the harmful side-effects of CRS and / or NT, e.g., CRS and / or NT associated with anti-CD3e targeting.

[0354] In some embodiments, the anti-TCRβV antibody molecule binds to one or more of TRBV2,TRBV3-1, TRBV4-1, TRBV4-2, TRBV4-3, TRBV5-1, TRBV5-4, TRBV5-5, TRBV5-6, TRBV5-8, TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8, TRBV6-9, TRBV7-2, TRBV7-3, TRBV7-4, TRBV7-6, TRBV7-7, TRBV7-8, TRBV7-9, TRBV9, TRBV10-1, TRBV10-2, TRBV10-3, TRBV11-1, TRBV11-2, TRBV11-3, TRBV12-3, TRBV12-4, TRBV12-5, TRBV13, TRBV14, TRBV15, TRBV16, TRBV18, TRBV19, TRBV20-1, TRBV24-1, TRBV25-1, TRBV27,WSGR Docket No.53676-768.601 TRBV28, TRBV29-1 and TRBV30. In some embodiments, the anti-TCRβV antibody molecule binds to one or more of TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8 and TRBV6-9. In some embodiments, the anti-TCRβV antibody molecule is an anti-TRBV2, anti-TRBV3-1, anti-TRBV4-1, anti-TRBV4-2, anti-TRBV4-3, anti-TRBV5-1, anti-TRBV5-4, anti-TRBV5-5, anti- TRBV5-6, anti-TRBV5-8, anti-TRBV6-1, anti-TRBV6-2, anti-TRBV6-3, anti-TRBV6-4, anti-TRBV6-5, anti-TRBV6-6, anti-TRBV6-8, anti-TRBV6-9, anti-TRBV7-2, anti-TRBV7-3, anti-TRBV7-4, anti- TRBV7-6, anti-TRBV7-7, anti-TRBV7-8, anti-TRBV7-9, anti-TRBV9, anti-TRBV10-1, anti-TRBV10- 2, anti-TRBV10-3, anti-TRBV11-1, anti-TRBV11-2, anti-TRBV11-3, anti-TRBV12-3, anti-TRBV12-4, anti-TRBV12-5, anti-TRBV13, anti-TRBV14, anti-TRBV15, anti-TRBV16, anti-TRBV18, anti- TRBV19, anti-TRBV20-1, anti-TRBV24-1, anti-TRBV25-1, anti-TRBV27, anti-TRBV28, anti- TRBV29-1, or anti-TRBV30. Exemplary anti-TCRβV antibody molecules and the corresponding TCRβV subfamilies recognized by said anti-TCRβV antibody molecules are disclosed in Table 10A.

[0355] In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV2,TRBV3-1, TRBV4-1, TRBV4-2, TRBV4-3, TRBV5-1, TRBV5-4, TRBV5-5, TRBV5-6, TRBV5-8, TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8, TRBV6-9, TRBV7-2, TRBV7-3, TRBV7-4, TRBV7-6, TRBV7-7, TRBV7-8, TRBV7-9, TRBV9, TRBV10-1, TRBV10-2, TRBV10-3, TRBV11-1, TRBV11-2, TRBV11-3, TRBV12-3, TRBV12-4, TRBV12-5, TRBV13, TRBV14, TRBV15, TRBV16, TRBV18, TRBV19, TRBV20-1, TRBV24-1, TRBV25-1, TRBV27, TRBV28, TRBV29-1 or TRBV30. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-1. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-2. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-3. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-4. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-5. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-6. In some embodiments, the anti- TCRβV antibody molecule binds specifically to TRBV6-8. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-9.

[0356] In some embodiments, the anti-TCRβV antibody molecule does not bind to TCRβ V12, or bindsto TCRβ V12 with an affinity and / or binding specificity that is less than (e.g., less than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0357] In some embodiments, the anti-TCRβV antibody molecule binds to TCRβ V12 with an affinityand / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0358] In some embodiments, the anti-TCRβV antibody molecule binds to a TCRβV region other thanTCRβ V12 (e.g., TCRβV region as described herein, e.g., TCRβ V6 subfamily (e.g., TCRβ V6-5*01) with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%,WSGR Docket No.53676-768.601 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0359] In some embodiments, the anti-TCRβV antibody molecule does not comprise the CDRs of theAntibody B murine antibody.

[0360] In some embodiments, the anti-TCRβV antibody molecule does not bind to TCRβ V5-5*01 orTCRβ V5-1*01, or binds to TCRβ V5-5*01 or TCRβ V5-1*01 with an affinity and / or binding specificity that is less than (e.g., less than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0361] In some embodiments, the anti-TCRβV antibody molecule binds to TCRβ V5-5*01 or TCRβ V5-1*01with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0362] In some embodiments, the anti-TCRβV antibody molecule binds to a TCRβV region other thanTCRβ V5-5*01 or TCRβ V5-1*01 (e.g., TCRβV region as described herein, e.g., TCRβ V6 subfamily (e.g., TCRβ V6-5*01) with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0363] In some embodiments, the anti-TCRβV antibody molecule does not comprise the CDRs of theTM23 murine antibody.

[0364] In some embodiments, the light or the heavy chain variable framework (e.g., the regionencompassing at least FR1, FR2, FR3, and optionally FR4) of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule can be chosen from: (a) a light or heavy chain variable framework including at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98%, or 100% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (b) a light or heavy chain variable framework including from 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (c) a non-human framework (e.g., a rodent framework); or (d) a non-human framework that has been modified, e.g., to remove antigenic or cytotoxic determinants, e.g., deimmunized, or partially humanized. In some embodiments, the light or heavy chain variable framework region (particularly FR1, FR2 and / or FR3) includes a light or heavy chain variable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94, 95, 96, 97, 98, 99% identical or identical to the frameworks of a VL or VH segment of a human germline gene.

[0365] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a heavy chain variable domain having at least one, two, three,WSGR Docket No.53676-768.601 four, five, six, seven, ten, fifteen, twenty or more changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the amino acid sequence of the FR region in the entire variable region, e.g., SEQ ID NO: 9.

[0366] Alternatively, or in combination with the heavy chain substitutions described herein, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the amino acid sequence of the FR region in the entire variable region, e.g., SEQ ID NO: 10 or SEQ ID NO: 11.

[0367] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes one, two, three, or four heavy chain framework regions, or a sequence substantially identical thereto.

[0368] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes one, two, three, or four light chain framework regions, or a sequence substantially identical thereto.

[0369] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the light chain framework region 1 of A-H.1 or A-H.2.

[0370] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the light chain framework region 2 of A-H.1 or A-H.2.

[0371] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the light chain framework region 3 of A-H.1 or A-H.2.

[0372] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the light chain framework region 4 of A-H.1 or A-H.2.

[0373] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 1 (FR1), comprising a change, e.g., a substitution (e.g., a conservative substitution) at position 10 according to Kabat numbering. In some embodiments, the FR1 comprises a Phenylalanine at position 10, e.g., a Serine to Phenylalanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0374] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 2 (FR2), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR2 comprises a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution. In some embodiments, FR2 comprises an Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., an Arginine to Alanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.WSGR Docket No.53676-768.601

[0375] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 3 (FR3), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR3 comprises a Phenylalanine at position 87, e.g., a substitution at position 87 according to Kabat numbering, e.g., a Tyrosine to Phenylalanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0376] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 1 (FR1) comprising a Phenylalanine at position 10, e.g., a substitution at position 10 according to Kabat numbering, e.g., a Serine to Phenylalanine substitution; (b) a framework region 2 (FR2) comprising a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution, and a Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., a Arginine to Alanine substitution; and (c) a framework region 3 (FR3) comprising a Phenylalanine at position 87, e.g., a substitution at position 87 according to Kabat numbering, e.g., a Tyrosine to Phenylalanine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 10. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0377] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 2 (FR2) comprising a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution, and a Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., a Arginine to Alanine substitution; and (b) a framework region 3 (FR3) comprising a Phenylalanine at position 87, e.g., a substitution at position 87 according to Kabat numbering, e.g., a Tyrosine to Phenylalanine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 11. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0378] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 1 (FR1) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) positions as described herein according to Kabat numbering, ; (b) a framework region 2 (FR2) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) position as described herein according to Kabat numbering and (c) a framework region 3 (FR3) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) position as described herein according to Kabat numbering. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0379] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the heavy chain framework region 1 of A-H.1 or A-H.2. In someWSGR Docket No.53676-768.601 embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 2 of A-H.1 or A-H.2, e.g. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 3 of A-H.1 or A-H.2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 4 of A-H.1 or A-H.2.

[0380] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a heavy chain variable domain comprising a framework region, e.g., framework region 3 (FR3), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR3 comprises a Threonine at position 73, e.g., a substitution at position 73 according to Kabat numbering, e.g., a Glutamic Acid to Threonine substitution. In some embodiments, FR3 comprises a Glycine at position 94, e.g., a substitution at position 94 according to Kabat numbering, e.g., an Arginine to Glycine substitution. In some embodiments, the substitution is relative to a human germline heavy chain framework region sequence.

[0381] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a heavy chain variable domain comprising a framework region 3 (FR3) comprising a Threonine at position 73, e.g., a substitution at position 73 according to Kabat numbering, e.g., a Glutamic Acid to Threonine substitution, and a Glycine at position 94, e.g., a substitution at position 94 according to Kabat numbering, e.g., a Arginine to Glycine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 10.

[0382] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.1 or A-H.2, e.g., SEQ ID NO: 9. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 10. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 11. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 9; and the light chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 10. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 9; and the light chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 11.

[0383] In some embodiments, the heavy or light chain variable domain, or both, of the anti-TCRβVantibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes an amino acid sequence, which is substantially identical to an amino acid as described herein, e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical to a variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or asWSGR Docket No.53676-768.601 described in Table 1, or encoded by the nucleotide sequence in Table 1; or which differs at least 1 or 5 residues, but less than 40, 30, 20, or 10 residues, from a variable region of an antibody described herein.

[0384] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises at least one, two, three, or four antigen-binding regions, e.g., variable regions, having an amino acid sequence as set forth in Table 1, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the sequences shown in Table 1. In another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes a VH and / or VL domain encoded by a nucleic acid having a nucleotide sequence as set forth in Table 1, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 1.

[0385] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 9, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 9; and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 10, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 10.

[0386] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 9, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 9; and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 11, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 11.

[0387] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule is a full antibody or fragment thereof (e.g., a Fab, F(ab')2, Fv, single domain antibody, or a single chain Fv fragment (scFv)). In embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule is a monoclonal antibody or an antibody with single specificity. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can also be a humanized, chimeric, camelid, shark, or an in vitro-generated antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, is a humanized antibody molecule. The heavy and light chains of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can be full-length (e.g., an antibody can include at least one,WSGR Docket No.53676-768.601 and preferably two, complete heavy chains, and at least one, and preferably two, complete light chains) or can include an antigen-binding fragment (e.g., a Fab, F(ab')2, Fv, a single chain Fv fragment, a single domain antibody, a diabody (dAb), a bivalent antibody, or bispecific antibody or fragment thereof, a single domain variant thereof, or a camelid antibody).

[0388] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, is in the form of a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0389] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, has a heavy chain constant region (Fc) chosen from, e.g., the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE. In some embodiments, the Fc region is chosen from the heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4. In some embodiments, the Fc region is chosen from the heavy chain constant region of IgG1 or IgG2 (e.g., human IgG1, or IgG2). In some embodiments, the heavy chain constant region is human IgG1. In some embodiments, the Fc region comprises a Fc region variant, e.g., as described herein.

[0390] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, has a light chain constant region chosen from, e.g., the light chain constant regions of kappa or lambda, preferably kappa (e.g., human kappa). In some embodiments, the constant region is altered, e.g., mutated, to modify the properties of the anti-TCRβV antibody molecule, e.g., anti- TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function). For example, the constant region is mutated at positions 296 (M to Y), 298 (S to T), 300 (T to E), 477 (H to K) and 478 (N to F) to alter Fc receptor binding (e.g., the mutated positions correspond to positions 132 (M to Y), 134 (S to T), 136 (T to E), 313 (H to K) and 314 (N to F) of SEQ ID NOs: 212 or 214; or positions 135 (M to Y), 137 (S to T), 139 (T to E), 316 (H to K) and 317 (N to F) of SEQ ID NOs: 215, 216, 217 or 218), e.g., relative to human IgG1.

[0391] Antibody A-H.1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:3278 and a light chain comprising the amino acid sequence of SEQ ID NO: 72. Antibody A-H.2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3278 and a light chain comprising the amino acid sequence of SEQ ID NO: 3279. Antibody A-H.68 comprises the amino acid sequence of SEQ ID NO: 1337, or a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto. Antibody A-H.69 comprises the amino acid sequence of SEQ ID NO: 1500, or a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto.

[0392] Additional exemplary humanized anti-TCRB V6 antibodies are provided in Table 1. In someembodiments, the anti-TCRβ V6 is antibody A, e.g., humanized antibody A (antibody A-H), as provided in Table 1. In some embodiments, the anti-TCRβV antibody comprises one or more (e.g., all three) of a LC CDR1, LC CDR2, and LC CDR3 provided in Table 1; and / or one or more (e.g., all three) of a HC CDR1, HC CDR2, and HC CDR3 provided in Table 1, or a sequence with at least 85%, 90%, 95%, 96%,WSGR Docket No.53676-768.601 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto. In some embodiments, antibody A comprises a heavy chain variable region (VH) and / or a light chain variable region (VL) provided in Table 1, or a sequence with at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto.

[0393] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a VH of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0394] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a VL of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0395] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a VH of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto; and a VL of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A-H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A- H.19, A-H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A- H.31, A-H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A- H.43, A-H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-WSGR Docket No.53676-768.601 H.55, A-H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A- H.67, A-H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A- H.79, A-H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0396] Exemplary anti-TCRβV antibody molecules and the corresponding TCRβV subfamiliesrecognized by said anti-TCRβV antibody molecules are disclosed in Table 10A.

[0397] The various TCRβV subfamilies and / or subfamily members can be expressed at different levelsin individuals, e.g., healthy individuals, as disclosed in Kitaura K. et al (2016), BMC Immunology vol 17: 38, the entire contents of which are hereby incorporated by reference. For example, TCRβ V6-5 is represented in approximately 3-6% healthy donors.

[0398] The representation of various TCRBV subfamilies and / or subfamily members can also bedifferent in cancer cells. For example, TCRβV is present in about 3-6% of tumor infiltrating T cells irrespective of tumor type (see Li B. et al., Nature Genetics, 2016, vol:48(7):725-32 the entire contents of which are hereby incorporated by references). Li et al., also disclose that TCRβ V6-5 is present at a high frequency in tumor cells. Anti-TCRβ V6 antibodies

[0399] In one aspect, provided herein is an anti-TCRβV antibody molecule that binds to human TCRβV6, e.g., a TCRβ V6 subfamily comprising: TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-9*01, TCRβ V6-8*01, TCRβ V6-5*01, TCRβ V6-6*02, TCRβ V6-6*01, TCRβ V6-2*01, TCRβ V6-3*01 or TCRβ V6-1*01. In some embodiments the TCRβ V6 subfamily comprises TCRβ V6-5*01 or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-9*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-8*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-5*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-2*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-3*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-1*01, or a variant thereof.

[0400] In some embodiments, TCRβ V6-5*01 is encoded by the nucleic acid sequence of SEQ ID NO:43, or a sequence having 85%, 90%, 95%, 99% or more identity thereof. In some embodiments, TCRβ V6-5*01 comprises the amino acid sequence of SEQ ID NO: 44, or an amino acid sequence having 85%, 90%, 95%, 99% or more identity thereof.

[0401] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, is a non-murine antibody molecule, e.g., a human or humanized antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti- TCRβ V6-5*01) antibody molecule is a human antibody molecule. In some embodiments, the anti-WSGR Docket No.53676-768.601 TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule is a humanized antibody molecule.

[0402] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, is isolated or recombinant.

[0403] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0404] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises at least one, two, three or four variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A- H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0405] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody molecule described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0406] In some embodiments, the anti-TCRβV antibody molecule comprises a heavy chain variableregion (VH) having a consensus sequence of SEQ ID NO: 231 or 3290.

[0407] SEQ ID NO: 231 - Consensus VHQVQLVQSGAEVKKPGSSVKVSCKASGH / T / G / YD / T / SFH / R / D / K / TL / D / K / T / NW / F / T / I / Y / GYIHWV RQAPGQGLEWMGR / WV / I / FF / S / YA / PGSGN / ST / V / Y / IK / RYNEKFKGRVTITADTSTSTAYMELSS LRSEDTAVYYCAG / VSY / IYSY / AD / GVLDYWGQGTTVTVSS

[0408] SEQ ID NO: 3290 - Consensus VHQVQLVQSGAEVKKPGSSVKVSCKASGX1X2FX3X4X5YIHWVRQAPGQGLEWMGX6X7X8X9G SGX10X11X12YNEKFKGRVTITADTSTSTAYMELSSLRSEDTAVYYCAX13SX14YSX15X16VLD YWGQGTTVTVSS, where-in: X1 is H or T or G or Y; X2 is D or T or S; X3 is H or R or D or K or T; X4 is L or D or K or T or N; X5 is W or F or T or I or Y or G; X6 is R or W; X7 is V or I or F; X8 is F or S or Y; X9 is A or P; X10 is N or S; X11 is T or V or Y or I; X12 is K or R; X13 is G or V; X14 is Y or I; X15 is Y or A; and X16 is D or G.

[0409] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-WSGR Docket No.53676-768.601 H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0410] In some embodiments, the anti-TCRβV antibody molecule comprises a light chain variableregion (VL) having a consensus sequence of SEQ ID NO: 230 or 3289.

[0411] SEQ ID NO: 230 - Consensus VLDIQMTQSPSFLSASVGDRVTITCKASQNVG / E / A / DN / DR / KVAWY / HQQKPGKAPKALIYSSSHRY K / SGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQFKSYPLTFGQGTKLEIK

[0412] SEQ ID NO: 3289 - Consensus VLDIQMTQSPSFLSASVGDRVTITCKASQNVX1X2X3VAWX4QQKPGKAPKALIYSSSHRYX5GVPS RFSGSGSGTEFTLTISSLQPEDFATYFCQQFKSYPLTFGQGTKLEIK, wherein X1 is G, E, A or D; X2 is N or D; X3 is R or K; X4 is Y or H; and X5 is K or S

[0413] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, comprises a heavy chain constant region for an IgG4, e.g., a human IgG4. In still another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes a heavy chain constant region for an IgG1, e.g., a human IgG1. In some embodiments, the heavy chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0414] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes a kappa light chain constant region, e.g., a human kappa light chain constant region. In some embodiments, the light chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0415] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region (VH) of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0416] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a heavy chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.WSGR Docket No.53676-768.601

[0417] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0418] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a light chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

[0419] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, three, four, five or six CDRs (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

[0420] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, molecule includes all six CDRs from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or closely related CDRs, e.g., CDRs which are identical or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein.

[0421] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 1) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A- H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 1.WSGR Docket No.53676-768.601

[0422] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 1) from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A- H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 1.

[0423] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, three, four, five, or six CDRs according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Kabat definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Kabat et al. shown in Table 1.

[0424] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes all six CDRs according to Kabat et al. (e.g., all six CDRs according to the Kabat definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A- H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Kabat et al. shown in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein.

[0425] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody chosen from chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the same canonical structures as at least loop 1 and / or loop 2 of the heavy and / or light chain variable domains of an antibody described herein. See, e.g., Chothia et al., (1992) J. Mol. Biol.227:799-817; Tomlinson et al., (1992) J. Mol. Biol.227:776-798 for descriptions of hypervariable loop canonicalWSGR Docket No.53676-768.601 structures or by ImMunoGeneTics (IMGT) system. These structures can be determined by inspection of the tables described in these references.

[0426] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 1) or by ImMunoGeneTics (IMGT) system from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or as described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 1 or by ImMunoGeneTics (IMGT) system.

[0427] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 1) or by ImMunoGeneTics (IMGT) system from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 1 or by ImMunoGeneTics (IMGT) system.

[0428] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes at least one, two, three, four, five, or six CDRs according to Chothia et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Chothia definition as set out in Table 1) or by ImMunoGeneTics (IMGT) system from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Chothia et al. shown in Table 1 or by ImMunoGeneTics (IMGT) system.

[0429] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, includes all six CDRs according to Chothia et al. (e.g., all six CDRs according to the Chothia definition as set out in Table 1) or by ImMunoGeneTics (IMGT) system from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, orWSGR Docket No.53676-768.601 encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Chothia et al. shown in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein or by ImMunoGeneTics (IMGT) system.

[0430] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, molecule includes a combination of CDRs or hypervariable loops defined according to Kabat et al., Chothia et al., or as described in Table 1 or by ImMunoGeneTics (IMGT) system.

[0431] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, can contain any combination of CDRs or hypervariable loops according to the Kabat and Chothia definitions or by ImMunoGeneTics (IMGT) system.

[0432] In some embodiments, a combined CDR as set out in Table 1 is a CDR that comprises a KabatCDR and a Chothia CDR or by ImMunoGeneTics (IMGT) system.

[0433] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule, molecule includes a combination of CDRs or hypervariable loops identified as combined CDRs in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can contain any combination of CDRs or hypervariable loops according the “combined” CDRs are described in Table 1.

[0434] In some embodiments, e.g., an embodiment comprising a variable region, a CDR (e.g., acombined CDR, Chothia CDR or Kabat CDR or CDRs by ImMunoGeneTics (IMGT) system), or other sequence referred to herein, e.g., in Table 1, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, a bivalent antibody molecule, a biparatopic antibody molecule, or an antibody molecule that comprises an antigen binding fragment of an antibody, e.g., a half antibody or antigen binding fragment of a half antibody. In certain embodiments the antibody molecule comprises a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0435] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule includes: (i) one, two or all of a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3) of SEQ ID NO: 2, SEQ ID NO: 10 or SEQ ID NO: 11, and / or (ii) one, two or all of a heavy chain complementarity determining region 1 (HC CDR1), heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3) of SEQ ID NO: 1 or SEQ ID NO: 9.

[0436] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 2, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 1.WSGR Docket No.53676-768.601

[0437] In some embodiments the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 10, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 9.

[0438] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 11, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 9.

[0439] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 6, a LC CDR2 amino acid sequence of SEQ ID NO: 7, or a LC CDR3 amino acid sequence of SEQ ID NO: 8; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 3, a HC CDR2 amino acid sequence of SEQ ID NO: 4, or a HC CDR3 amino acid sequence of SEQ ID NO: 5.

[0440] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 6, a LC CDR2 amino acid sequence of SEQ ID NO: 7, or a LC CDR3 amino acid sequence of SEQ ID NO: 8; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 3, a HC CDR2 amino acid sequence of SEQ ID NO: 4, or a HC CDR3 amino acid sequence of SEQ ID NO: 5.

[0441] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 51, a LC CDR2 amino acid sequence of SEQ ID NO: 52, or a LC CDR3 amino acid sequence of SEQ ID NO: 53; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 45, a HC CDR2 amino acid sequence of SEQ ID NO: 46, or a HC CDR3 amino acid sequence of SEQ ID NO: 47.

[0442] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 51, a LC CDR2 amino acid sequence of SEQ ID NO: 52, or a LC CDR3 amino acid sequence of SEQ ID NO: 53; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 45, a HC CDR2 amino acid sequence of SEQ ID NO: 46, or a HC CDR3 amino acid sequence of SEQ ID NO: 47.

[0443] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 54, a LC CDR2 amino acid sequence of SEQ ID NO: 55, or a LC CDR3 amino acid sequence of SEQ ID NO: 56; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 48, a HC CDR2 amino acid sequence of SEQ ID NO: 49, or a HC CDR3 amino acid sequence of SEQ ID NO: 50.

[0444] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 54, a LC CDR2 amino acid sequence of SEQ ID NO: 55, or a LC CDR3 amino acid sequence of SEQ ID NO: 56; and / or (ii) a heavy chain variable region (VH)WSGR Docket No.53676-768.601 comprising a HC CDR1 amino acid sequence of SEQ ID NO: 48, a HC CDR2 amino acid sequence of SEQ ID NO: 49, or a HC CDR3 amino acid sequence of SEQ ID NO: 50.

[0445] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a VH and / or a VL of an antibody described in Table 1, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0446] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβV6-5*01) antibody molecule comprises a VH and a VL of an antibody described in Table 1, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0447] In some embodiments, an anti-TCRVb antibody as described herein has an antigen bindingdomain having a VL having a consensus sequence of SEQ ID NO: 230, wherein position 30 is G, E, A or D; position 31 is N or D; position 32 is R or K; position 36 is Y or H; and / or position 56 is K or S.

[0448] In some embodiments, an anti-TCRVb antibody as described herein has an antigen bindingdomain having a VH having a consensus sequence of SEQ ID NO: 231, wherein: position 27 is H or T or G or Y; position 28 is D or T or S; position 30 is H or R or D or K or T; position 31 is L or D or K or T or N; position 32 is W or F or T or I or Y or G; position 49 is R or W; position 50 is V or I or F; position 51 is F or S or Y; position 52 is A or P; position 56 is N or S; position 57 is T or V or Y or I; position 58 is K or R; position 97 is G or V; position 99 is Y or I; position 102 is Y or A; and / or position 103 is D or G. Anti-TCRβ V12 antibodies

[0449] In one aspect, provided herein is an anti-TCRβV antibody molecule that binds to human TCRβV12, e.g., a TCRβ V12 subfamily comprising: TCRβ V12-4*01, TCRβ V12-3*01 or TCRβ V12-5*01. In some embodiments the TCRβ V12 subfamily comprises TCRβ V12-4*01. In some embodiments the TCRβ V12 subfamily comprises TCRβ V12-3*01.

[0450] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, is a non-murine antibody molecule, e.g., a human or humanized antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule is a human antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule is a humanized antibody molecule.

[0451] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, is isolated or recombinant.

[0452] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0453] In some embodiments, the anti- antibody molecule,anti-TCRβ V12 antibodymolecule, comprises at least one, two, three or four variable regions from an antibody described herein,WSGR Docket No.53676-768.601 e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0454] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0455] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0456] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, comprises a heavy chain constant region for an IgG4, e.g., a human IgG4. In still another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes a heavy chain constant region for an IgG1, e.g., a human IgG1. In some embodiments, the heavy chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0457] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, includes a kappa light chain constant region, e.g., a human kappa light chain constant region. In some embodiments, the light chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0458] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, includes at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0459] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a heavy chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0460] In some embodiments, the anti-TCRβV antibody molecule,anti-TCRβ V12 antibodymolecule, includes at least one, two, or three complementarity determining regions (CDRs) from a lightWSGR Docket No.53676-768.601 chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0461] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a light chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0462] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, includes at least one, two, three, four, five or six CDRs (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0463] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, molecule includes all six CDRs from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2, or closely related CDRs, e.g., CDRs which are identical or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, may include any CDR described herein.

[0464] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 2) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 2.

[0465] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 2) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three orWSGR Docket No.53676-768.601 four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 2.

[0466] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, three, four, five, or six CDRs according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Kabat definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Kabat et al. shown in Table 2.

[0467] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes all six CDRs according to Kabat et al. (e.g., all six CDRs according to the Kabat definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Kabat et al. shown in Table 2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V12 antibody molecule may include any CDR described herein.

[0468] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody described in Table 2, e.g., the same canonical structures as at least loop 1 and / or loop 2 of the heavy and / or light chain variable domains of an antibody described herein. See, e.g., Chothia et al., (1992) J. Mol. Biol.227:799- 817; Tomlinson et al., (1992) J. Mol. Biol.227:776-798 for descriptions of hypervariable loop canonical structures. These structures can be determined by inspection of the tables described in these references.

[0469] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 2) or by ImMunoGeneTics (IMGT) system from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 2 or by ImMunoGeneTics (IMGT) system.WSGR Docket No.53676-768.601

[0470] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 2) or by ImMunoGeneTics (IMGT) system from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 2 or by ImMunoGeneTics (IMGT) system.

[0471] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, three, four, five, or six CDRs according to Chothia et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Chothia definition as set out in Table 2) or by ImMunoGeneTics (IMGT) system from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Chothia et al. shown in Table 2 or by ImMunoGeneTics (IMGT) system.

[0472] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes all six CDRs according to Chothia et al. (e.g., all six CDRs according to the Chothia definition as set out in Table 2) or by ImMunoGeneTics (IMGT) system from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Chothia et al. shown in Table 2 or by ImMunoGeneTics (IMGT) system. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule may include any CDR described herein.

[0473] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to a combined CDR (e.g., at least one, two, or three CDRs according to the combined CDR definition as set out in Table 2) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to combined CDR shown in Table 2.WSGR Docket No.53676-768.601

[0474] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, or three CDRs according to a combined CDR (e.g., at least one, two, or three CDRs according to the combined CDR definition as set out in Table 2) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to a combined CDR shown in Table 2.

[0475] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes at least one, two, three, four, five, or six CDRs according to a combined CDR. (e.g., at least one, two, three, four, five, or six CDRs according to the combined CDR definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to a combined CDR shown in Table 2.

[0476] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes all six CDRs according to a combined CDR (e.g., all six CDRs according to the combined CDR definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to a combined CDR shown in Table 2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule may include any CDR described herein.

[0477] In some embodiments, a combined CDR as set out in Table 1 is a CDR that comprises a KabatCDR and a Chothia CDR or CDR by ImMunoGeneTics (IMGT) system.

[0478] In some embodiments, the anti-TCRβV antibody molecule, e e.g., anti-TCRβ V12 antibodymolecule, molecule includes a combination of CDRs or hypervariable loops identified as combined CDRs in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, can contain any combination of CDRs or hypervariable loops according the “combined” CDRs are described in Table 1.

[0479] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes a combination of CDRs or hypervariable loops defined according to the Kabat et al. and Chothia et al., or as described in Table 1 or by ImMunoGeneTics (IMGT) system.WSGR Docket No.53676-768.601

[0480] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule can contain any combination of CDRs or hypervariable loops according to the Kabat and Chothia definitions or by ImMunoGeneTics (IMGT) system.

[0481] In some embodiments, e.g., an embodiment comprising a variable region, a CDR (e.g., acombined CDR, Chothia CDR or Kabat CDR, or CDR by ImMunoGeneTics (IMGT) system), or other sequence referred to herein, e.g., in Table 2, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, a bivalent antibody molecule, a biparatopic antibody molecule, or an antibody molecule that comprises an antigen binding fragment of an antibody, e.g., a half antibody or antigen binding fragment of a half antibody. In certain embodiments the antibody molecule comprises a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0482] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes: (i) one, two or all of a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3) of SEQ ID NO: 16, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29 or SEQ ID NO: 30, and / or (ii) one, two or all of a heavy chain complementarity determining region 1 (HC CDR1), heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3) of SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 24 or SEQ ID NO: 25.

[0483] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 20, a LC CDR2 amino acid sequence of SEQ ID NO: 21, or a LC CDR3 amino acid sequence of SEQ ID NO: 22; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 17, a HC CDR2 amino acid sequence of SEQ ID NO: 18, or a HC CDR3 amino acid sequence of SEQ ID NO: 19.

[0484] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 20, a LC CDR2 amino acid sequence of SEQ ID NO: 21, and a LC CDR3 amino acid sequence of SEQ ID NO: 2; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 17, a HC CDR2 amino acid sequence of SEQ ID NO: 18, and a HC CDR3 amino acid sequence of SEQ ID NO: 19.

[0485] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0486] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1WSGR Docket No.53676-768.601 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0487] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 66, a LC CDR2 amino acid sequence of SEQ ID NO: 67, or a LC CDR3 amino acid sequence of SEQ ID NO: 68; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 60, a HC CDR2 amino acid sequence of SEQ ID NO: 61, or a HC CDR3 amino acid sequence of SEQ ID NO: 62.

[0488] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0489] In some embodiments, the light or the heavy chain variable framework (e.g., the regionencompassing at least FR1, FR2, FR3, and optionally FR4) of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule can be chosen from: (a) a light or heavy chain variable framework including at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98%, or 100% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (b) a light or heavy chain variable framework including from 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (c) a non-human framework (e.g., a rodent framework); or (d) a non-human framework that has been modified, e.g., to remove antigenic or cytotoxic determinants, e.g., deimmunized, or partially humanized. In some embodiments, the light or heavy chain variable framework region (particularly FR1, FR2 and / or FR3) includes a light or heavy chain variable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94, 95, 96, 97, 98, 99% identical or identical to the frameworks of a VL or VH segment of a human germline gene.

[0490] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule, comprises a heavy chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more changes, e.g., amino acid substitutions or deletions, from an amino acid sequence described in Table 2.e.g., the amino acid sequence of the FR region in the entire variable region, e.g., shown in FIGs.3A and 3B, or in SEQ ID NOs: 23-25.

[0491] Alternatively, or in combination with the heavy chain substitutions described herein the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of an antibodyWSGR Docket No.53676-768.601 described herein .e.g., the amino acid sequence of the FR region in the entire variable region, e.g., shown in FIGs.3A and 3B, or in SEQ ID NOs: 26-30.

[0492] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes one, two, three, or four heavy chain framework regions shown in FIG.3A, or a sequence substantially identical thereto.

[0493] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule includes one, two, three, or four light chain framework regions shown in FIG.3B, or a sequence substantially identical thereto. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 1 e.g., as shown in FIG. 3B. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 2 e.g., as shown in FIG.3B. In some embodiments, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 3, e.g., as shown in FIG.3B. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V12 antibody molecule comprises the light chain framework region 4, e.g., as shown in FIG.3B.

[0494] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more, e.g., all, position as described herein according to Kabat numbering. In some embodiments, FR1 comprises an Aspartic Acid at position 1, e.g., a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution. In some embodiments, FR1 comprises an Asparagine at position 2, e.g., a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution. In some embodiments, FR1 comprises a Leucine at position 4, e.g., a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution.

[0495] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution, a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution, and a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution, and a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution. In some embodiments, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution, and a substitution at position 4WSGR Docket No.53676-768.601 according to Kabat numbering, e.g., a Methionine to Leucine substitution. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution, and a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0496] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising a framework region, e.g., framework region 3 (FR3), comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more, e.g., all, position as described herein according to Kabat numbering. In some embodiments, FR3 comprises a Glycine at position 66, e.g., a substitution at position 66 according to Kabat numbering, e.g., a Lysine to Glycine substitution, or a Serine to Glycine substitution. In some embodiments, FR3 comprises an Asparagine at position 69, e.g., a substitution at position 69 according to Kabat numbering, e.g., a Tyrosine to Asparagine substitution. In some embodiments, FR3 comprises a Tyrosine at position 71, e.g., a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, or an Alanine to Tyrosine substitution.

[0497] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising a framework region, e.g., framework region 3 (FR3), comprising a substitution at position 66 according to Kabat numbering, e.g., a Lysine to Glycine substitution, or a Serine to Glycine substitution, and a substitution at position 69 according to Kabat numbering, e.g., a Tyrosine to Asparagine substitution.. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 3 (FR3), comprising a substitution at position 66 according to Kabat numbering, e.g., Lysine to Glycine substitution, or a Serine to Glycine substitution, and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, or an Alanine to Tyrosine substitution. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 3 (FR3), comprising a substitution at position 69 according to Kabat numbering, e.g., a Tyrosine to Asparagine substitution and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, or an Alanine to Tyrosine substitution. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 3 (FR3), comprising a substitution at position 66 according to Kabat numbering, e.g., a Lysine to Glycine substitution, or a Serine to Glycine substitution, a substitution at position 69 according to Kabat numbering, e.g., a Tyrosine to Asparagine substitution and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, or an Alanine to Tyrosine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.WSGR Docket No.53676-768.601

[0498] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising: a framework region 1 (FR1) comprising a substitution at position 2 according to Kabat numbering, e.g., a Isoleucine to Asparagine substitution; and a framework region 3 (FR3), comprising a substitution at position 69 according to Kabat numbering, e.g., a Threonine to Asparagine substitution and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 26. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0499] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising: (a) a framework region 1 (FR1) comprising a substitution at position 1 according to Kabat numbering, e.g., a Alanine to Aspartic Acid substitution, and a substitution at position 2 according to Kabat numbering, e.g., a Isoleucine to Asparagine substitution; and (b) a framework region 3 (FR3), comprising a substitution at position 69 according to Kabat numbering, e.g., a Threonine to Asparagine substitution and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 27 In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0500] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising: (a) a framework region 1 (FR1) comprising a substitution at position 2 according to Kabat numbering, e.g., a Serine to Asparagine substitution; and a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution; and (b) a framework region 3 (FR3), comprising a substitution at position 69 according to Kabat numbering, e.g., a Threonine to Asparagine substitution and a substitution at position 71 according to Kabat numbering, e.g., a Phenylalanine to Tyrosine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 28 In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0501] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising: (a) a framework region 1 (FR1) comprising a substitution at position 2 according to Kabat numbering, e.g., a Serine to Asparagine substitution; and (b) a framework region 3 (FR3) comprising a substitution at position 66 according to Kabat numbering, e.g., a Lysine to Glycine substitution; a substitution at position 69 according to Kabat numbering, e.g., a Threonine to Asparagine substitution; and a substitution at position 71 according to Kabat numbering, e.g., a Alanine to Tyrosine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 29. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0502] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain comprising: (a) a framework region 1 (FR1) comprising a substitution at position 2 according to Kabat numbering, e.g., a Tyrosine to Asparagine substitution; and (b) aWSGR Docket No.53676-768.601 framework region 3 (FR3) comprising a substitution at position 66 according to Kabat numbering, e.g., a Serine to Glycine substitution; a substitution at position 69 according to Kabat numbering, e.g., a Threonine to Asparagine substitution; and a substitution at position 71 according to Kabat numbering, e.g., a Alanine to Tyrosine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 29. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0503] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises a light chain variable domain comprising: (a) a framework region 1 (FR1) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) positions as described herein according to Kabat numbering, and (b) a framework region 3 (FR3) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) position as described herein according to Kabat numbering. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0504] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises the heavy chain framework region 1, e.g., as shown in FIG.3A. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the heavy chain framework region 2, e.g., as shown in FIG.3A. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the heavy chain framework region 3, e.g., as shown in FIG.3A. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the heavy chain framework region 4, e.g., as shown in FIG.3A. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the heavy chain framework regions 1-4, e.g., SEQ ID NOS: 20-23, or as shown in FIG.3A. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework regions 1-4, e.g., SEQ ID NOs: 26-30, or as shown in FIG.3B.

[0505] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises the heavy chain framework regions 1-4, e.g., SEQ ID NOs: 23-25; and the light chain framework regions 1-4, e.g., SEQ ID NOs: 26-30, or as shown in FIGs.3A and 3B.

[0506] In some embodiments, the heavy or light chain variable domain, or both, of , the anti-TCRβVantibody molecule, e.g., anti-TCRβ V12 antibody molecule includes an amino acid sequence, which is substantially identical to an amino acid as described herein, e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical to a variable region of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or which differs at least 1 or 5 residues, but less than 40, 30, 20, or 10 residues, from a variable region of an antibody described herein.

[0507] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises at least one, two, three, or four antigen-binding regions, e.g., variable regions, having an amino acid sequence as set forth in Table 2, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the sequences shown in Table 2. In another embodiment, , theWSGR Docket No.53676-768.601 anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes a VH and / or VL domain encoded by a nucleic acid having a nucleotide sequence as set forth in Table 2, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 2.

[0508] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising an amino acid sequence chosen from the amino acid sequence of SEQ ID NO: 23, SEQ ID NO:24 or SEQ ID NO:25, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, SEQ ID NO:24 or SEQ ID NO:25, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23, SEQ ID NO:24 or SEQ ID NO:25; and / or a VL domain comprising an amino acid sequence chosen from the amino acid sequence of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29 or SEQ ID NO: 30, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29 or SEQ ID NO: 30, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29 or SEQ ID NO: 30. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 23, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23; and a VL domain comprising the amino acid sequence of SEQ ID NO: 26, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 26, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 26.

[0509] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 23, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23; and a VL domain comprising the amino acid sequence of SEQ ID NO: 27, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 27, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 27.

[0510] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 23, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23; and a VL domain comprising the amino acid sequenceWSGR Docket No.53676-768.601 of SEQ ID NO: 28, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 28, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 28.

[0511] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 23, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23; and a VL domain comprising the amino acid sequence of SEQ ID NO: 29, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 29, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 29.

[0512] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 23, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 23, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 23; and a VL domain comprising the amino acid sequence of SEQ ID NO: 30, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 30, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 30.

[0513] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 24 or 25, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 24 or 25, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 24 or 25; and a VL domain comprising the amino acid sequence of SEQ ID NO: 26, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence SEQ ID NO: 26, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 26.

[0514] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibodymolecule comprises: a VH domain comprising the amino acid sequence of SEQ...

Claims

WSGR Docket No.53676-768.601 CLAIMS WHAT IS CLAIMED IS:

1. A multifunctional molecule comprising:(a) a first domain that binds to a first target molecule, wherein the first target molecule is a T cellreceptor alpha (TCRα) chain; and (b) a second domain that binds to a second target molecule, wherein the second target molecule isa T cell receptor beta (TCRβ) chain.

2. The multifunctional molecule of claim 1, wherein the TCRα chain is a human TCRα.

3. The multifunctional molecule of claim 1 or 2, wherein the TCRβ chain is a human TCRβ chain.

4. The multifunctional molecule of any one of claims 1-3, wherein the first domain is a TCRαV-bindingdomain that binds to a variable region of the human TCRα chain (TCRαV).

5. The multifunctional molecule of any one of claims 1-4, wherein the second domain is a TCRβV-binding domain that binds to a variable region of the human TCRβ chain (TCRβV).

6. The multifunctional molecule of any one of claims 1-5, wherein the multifunctional moleculecomprises at least two non-contiguous polypeptides comprising a first polypeptide chain and a second polypeptide chain; wherein the first polypeptide chain comprises a first portion of a dimerization module; wherein the second polypeptide chain comprises a second portion of the dimerization module; and wherein the first polypeptide chain and the second polypeptide chain form a dimer via association of the first portion of the dimerization module and the second portion of the dimerization module.

7. The multifunctional molecule of any one of claims 1-6, wherein the first domain is an antibodymolecule.

8. The multifunctional molecule of any one of claims 1-7, wherein the second domain is an antibodymolecule.

9. The multifunctional molecule of claim 7 or 8, wherein the antibody molecule is selected from thegroup consisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

10. The multifunctional molecule of any one of claims 7-9, wherein the first domain is a scFv or a singledomain antibody.

11. The multifunctional molecule of any one of claims 7-9, wherein the first domain is an Fab comprisinga first portion of the first domain and a second portion of the first domain; wherein the first portion of the first domain and the second portion of the first domain assemble and form the first domain of the multifunctional molecule;WSGR Docket No.53676-768.601 wherein the multifunctional molecule further comprises a polypeptide chain comprising the second portion of the first domain, wherein the polypeptide chain comprising the second portion of the first domain is non-contiguous with the first polypeptide chain and the second polypeptide chain.

12. The multifunctional molecule of claim 11, wherein the first portion of the first domain comprises aheavy chain variable region (VH) of the Fab and the second portion of the first domain comprises a light chain variable region (VL) of the Fab, or the first portion of the first domain comprises the VL of the Fab and the second portion of the first domain comprises the VH of the Fab.

13. The multifunctional molecule of any one of claims 7-12, wherein the second domain is a scFv or asingle domain antibody.

14. The multifunctional molecule of any one of claims 7-12, wherein the second domain is an Fabcomprising a first portion of the second domain and a second portion of the second domain; wherein the first portion of the second domain and the second portion of the second domain assemble and form the second domain of the multifunctional molecule; wherein the multifunctional molecule further comprises a polypeptide chain comprising the second portion of the second domain, wherein the polypeptide chain comprising the second portion of the second domain is non-contiguous with the first polypeptide chain, the second polypeptide chain, and the polypeptide chain comprising the second portion of the first domain.

15. The multifunctional molecule of claim 14, wherein the first portion of the second domain comprises aheavy chain variable region (VH) of the Fab and the second portion of the second domain comprises a light chain variable region (VL) of the Fab, or the first portion of the second domain comprises the VL of the Fab and the second portion of the second domain comprises the VH of the Fab.

16. The multifunctional molecule of any one of claims 6-15, wherein the first portion of the dimerizationmodule is linked to the first domain or the first portion of the first domain.

17. The multifunctional molecule of claim 16, wherein the N-terminus of the first portion of thedimerization module is linked to the C-terminus of the first domain or the C-terminus of the first portion of the first domain.

18. The multifunctional molecule of claim 16, wherein the C-terminus of the first portion of thedimerization module is linked to the N-terminus of the first domain or the N-terminus of the first portion of the first domain.

19. The multifunctional molecule of any one of claims 6-18, wherein the first portion of the dimerizationmodule is linked to the second domain or the first portion of the second domain.

20. The multifunctional molecule of any one of claims 6-19, wherein the N-terminus of the first portionof the dimerization module is linked to the C-terminus of the first domain or the C-terminus of the first portion of the first domain, and the C-terminus of the first portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

21. The multifunctional molecule of any one of claims 6-19, wherein the C-terminus of the first portionof the dimerization module is linked to the N-terminus of the first domain or the N-terminus of theWSGR Docket No.53676-768.601 first portion of the first domain, and the N-terminus of the first portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

22. The multifunctional molecule of any one of claims 6-18, wherein the second portion of thedimerization module is linked to the second domain or the first portion of the second domain.

23. The multifunctional molecule of claim 22, wherein the N-terminus of the second portion of thedimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

24. The multifunctional molecule of claim 22, wherein the C-terminus of the second portion of thedimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

25. The multifunctional molecule of any one of claims 6-18 and 22-24, wherein the N-terminus of thefirst portion of a dimerization module is linked to the C-terminus of the first domain or the C- terminus of the first portion of the first domain, and the N-terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

26. The multifunctional molecule of any one of claims 6-18 and 22-24, wherein the N-terminus of thefirst portion of the dimerization module is linked to the C-terminus of the first domain or the C- terminus of the first portion of the first domain, and the C-terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

27. The multifunctional molecule of any one of claims 6-18 and 22-24, wherein the C-terminus of thefirst portion of the dimerization module is linked to the N-terminus of the first domain or the N- terminus of the first portion of the first domain, and the C-terminus of the second portion of the dimerization module is linked to the N-terminus of the second domain or the N-terminus of the first portion of the second domain.

28. The multifunctional molecule of any one of claims 6-18 and 22-24, wherein the C-terminus of thefirst portion of the dimerization module is linked to the N-terminus of the first domain or the N- terminus of the first portion of the first domain, and the N-terminus of the second portion of the dimerization module is linked to the C-terminus of the second domain or the C-terminus of the first portion of the second domain.

29. The multifunctional molecule of claims 6-9, 11-13, and 15-28, wherein the multifunctional moleculecomprises at least three non-contiguous polypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to a first portion of the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module; and (iii) a polypeptide chain comprising a second portion of the first domain; andWSGR Docket No.53676-768.601 wherein the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

30. The multifunctional molecule of claims 6-10 and 14-28, wherein the multifunctional moleculecomprises at least three non-contiguous polypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module; and (iii) a polypeptide chain comprising a second portion of the second domain; and wherein a first portion of the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

31. The multifunctional molecule of claims 6-9, 11, 12, and 14-28, wherein the multifunctional moleculecomprises at least four non-contiguous polypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked to a first portion of the first domain; (ii) the second polypeptide chain comprising the second portion of the dimerization module; (iii) a polypeptide chain comprising a second portion of the first domain; and (iv) a polypeptide chain comprising a second portion of the second domain; and wherein a first portion of the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

32. The multifunctional molecule of claims 6-10, 13, and 16-28, wherein the multifunctional moleculecomprises at least two non-contiguous polypeptide chains, wherein: (i) the first polypeptide chain comprising the first portion of the dimerization module linked tothe first domain; and (ii) the second polypeptide chain comprising the second portion of the dimerization module; andwherein the second domain is linked to the first portion of the dimerization module or the second portion of the dimerization module.

33. The multifunctional molecule of any one of claims 1-32, wherein the multifunctional moleculefurther comprises at least one cytokine or a functional fragment or variant thereof.

34. The multifunctional molecule of claim 33, wherein the at least one cytokine or a functional fragmentor variant thereof is linked to the first portion of the dimerization module, the second portion of the dimerization module, or any combination thereof.

35. The multifunctional molecule of claim 33 or 34, wherein the at least one cytokine or a functionalfragment or variant thereof is linked to the N-terminus of the first portion of the dimerization module, the C-terminus of the first portion of the dimerization module, the N-terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof.

36. The multifunctional molecule of any one of claims 33-35, wherein the at least one cytokine or afunctional fragment or variant thereof is linked to the first domain, the first portion of the firstWSGR Docket No.53676-768.601 domain, the second portion of the first domain, the second domain, the first portion of the second domain, the second portion of the second domain, or any combination thereof.

37. The multifunctional molecule of claim 36, wherein the at least one cytokine or a functional fragmentor variant thereof is linked to the N-terminus of the first domain, the C-terminus of the first domain, the N-terminus of the first portion of the first domain, the C-terminus of the first portion of the first domain, the N-terminus of the second portion of the first domain, the C-terminus of the second portion of the first domain, the N-terminus of the second domain, the C-terminus of the second domain, the N-terminus of the first portion of the second domain, the C-terminus of the first portion of the second domain, the N-terminus of the second portion of the second domain, the C-terminus of the second portion of the second domain, or any combination thereof.

38. The multifunctional molecule of any one of claims 33-37, wherein the at least one cytokine or afunctional fragment or variant thereof is selected from the group consisting of interleukin-2 (IL-2) or functional variant thereof, interleukin-7 (IL-7) or functional variant thereof, interleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL-15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL-21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

39. The multifunctional molecule of claim 38, wherein the at least one cytokine or a functional fragmentor variant thereof comprises interleukin-2 (IL-2) or functional variant thereof.

40. The multifunctional molecule of claim 38 or 39, wherein the interleukin-2 (IL-2) or functional variantthereof comprises a sequence with at least 70% sequence identity to the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

41. The multifunctional molecule of any one of claims 38-40, wherein the interleukin-2 (IL-2) orfunctional variant thereof comprises the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

42. The multifunctional molecule of claim 38, wherein the at least one cytokine or a functional fragmentor variant thereof comprises interleukin-15 (IL-15) or functional variant thereof.

43. The multifunctional molecule of claim 38 or 42, wherein the interleukin-15 (IL-15) or functionalvariant thereof comprises a sequence with at least 70% sequence identity to the sequence of SEQ ID NO: 2170.

44. The multifunctional molecule of any one of claims 38, 42, and 43, wherein the interleukin-15 (IL-15)or functional variant thereof comprises the sequence of SEQ ID NO: 2170.

45. The multifunctional molecule of any one of claims 38 and 42-44, wherein the at least one cytokine ora functional fragment or variant thereof further comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

46. The multifunctional molecule of claim 45, wherein the IL15Ralpha dimerizing domain comprises anIL-15 receptor alpha sushi domain or functional variant thereof.

47. The multifunctional molecule of claim 45 or 46, wherein the IL15Ralpha dimerizing domaincomprises a sequence with at least 70% sequence identity to the sequence of SEQ ID NO: 3472.WSGR Docket No.53676-768.60148. The multifunctional molecule of any one of claims 45-47, wherein the IL15Ralpha dimerizingdomain comprises the sequence of SEQ ID NO: 3472.

49. The multifunctional molecule of any one of claims 45-48, wherein the interleukin-15 (IL-15) orfunctional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

50. The multifunctional molecule of claim 49, wherein the interleukin-15 (IL-15) or functional variantthereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

51. The multifunctional molecule of any one of claims 38 and 42-50, wherein the at least one cytokine ora functional fragment or variant thereof comprises a sequence with at least 70% sequence identity to the sequence of SEQ ID NO: 3474.

52. The multifunctional molecule of any one of claims 38 and 42-51, wherein the at least one cytokine ora functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3474.

53. The multifunctional molecule of any one of claims 1-52, wherein the multifunctional moleculefurther comprises a linker between the first domain or the first portion of the first domain and the first portion of the dimerization module.

54. The multifunctional molecule of any one of claims 1-53, wherein the multifunctional moleculefurther comprises a linker between the second domain or the first portion of the second domain and the first portion of a dimerization module.

55. The multifunctional molecule of any one of claims 1-54, wherein the multifunctional moleculefurther comprises a linker between the second domain or the first portion of the second domain and the second portion of a dimerization module.

56. The multifunctional molecule of any one of claims 33-55, wherein the multifunctional moleculefurther comprises a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the dimerization module.

57. The multifunctional molecule of any one of claims 33-56, wherein the multifunctional moleculefurther comprises a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the dimerization module of the first polypeptide chain.

58. The multifunctional molecule of any one of claims 33-57, wherein the multifunctional moleculefurther comprises a linker between the at least one cytokine or a functional fragment or variant thereof and the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second portion of the first domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the first portion of the second domain, a linker between the at least one cytokine or a functional fragment or variant thereof and the second portion of the second domain, or any combination thereof.

59. The multifunctional molecule of any one of claims 1-58, wherein the multifunctional moleculefurther comprises a tumor-targeting moiety.WSGR Docket No.53676-768.60160. The multifunctional molecule of claim 59, wherein the tumor-targeting moiety binds to a cancerantigen.

61. The multifunctional molecule of claim 59 or 60, wherein the tumor-targeting moiety binds to a cancerantigen selected from the group consisting of CD19, CD123, CD22, CD30, CD171, CS-1, C-type lectin-like molecule-1, CD33, epidermal growth factor receptor variant III (EGFRvIII), ganglioside G2 (GD2), ganglioside GD3, TNF receptor family member B cell maturation (BCMA), Tn antigen ((Tn Ag) or (GalNAcα-Ser / Thr)), prostate-specific membrane antigen (PSMA), Receptor tyrosine kinase-like orphan receptor 1 (ROR1), Fms-Like Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD117), Interleukin-13 receptor subunit alpha- 2, mesothelin, Interleukin 11 receptor alpha (IL-11Ra), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage-specific embryonic antigen-4 (SSEA-4), CD20, Folate receptor alpha, Receptor tyrosine-protein kinase ERBB2 (Her2 / neu), Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion molecule (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gp100) / pmel17, oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr- abl), tyrosinase, ephrin type-A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma-associated antigen (HMWMAA), o-acetyl-GD2 ganglioside (OAcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related (TEM7R), claudin 6 (CLDN6), thyroid stimulating hormone receptor (TSHR), G protein-coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, CD179a, anaplastic lymphoma kinase (ALK), Polysialic acid, placenta-specific 1 (PLAC1), hexasaccharide portion of globoH glycoceramide (GloboH), mammary gland differentiation antigen (NY-BR-1), uroplakin 2 (UPK2), Hepatitis A virus cellular receptor 1 (HAVCR1), adrenoceptor beta 3 (ADRB3), pannexin 3 (PANX3), G protein-coupled receptor 20 (GPR20), lymphocyte antigen 6 complex, locus K 9 (LY6K), Olfactory receptor 51E2 (OR51E2), TCR Gamma Alternate Reading Frame Protein (TARP), Wilms tumor protein (WT1), Cancer / testis antigen 1 (NY-ESO-1) / LAGE-1, Cancer / testis antigen 2 (LAGE-1a), Melanoma-associated antigen 1 (MAGE-A1), ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML), sperm protein 17 (SPA17), X Antigen Family, Member 1A (XAGE1), angiopoietin-binding cell surface receptor 2 (Tie 2), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), Fos-related antigen 1, tumor protein p53 (p53), p53 mutant, prostein, survivin, telomerase, prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1, Rat sarcoma (Ras) mutant, human TelomeraseWSGR Docket No.53676-768.601 reverse transcriptase (hTERT), sarcoma translocation breakpoints, melanoma inhibitor of apoptosis (ML-IAP), ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene), N-Acetyl glucosaminyl-transferase V (NA17), paired box protein Pax-3 (PAX3), Androgen receptor, Cyclin B1, v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN), Ras Homolog Family Member C (RhoC), Tyrosinase-related protein 2 (TRP-2), Cytochrome P4501B1 (CYP1B1), CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3), Paired box protein Pax-5 (PAX5), proacrosin binding protein sp32 (OY-TES1), lymphocyte-specific protein tyrosine kinase (LCK), A kinase anchor protein 4 (AKAP-4), synovial sarcoma, X breakpoint 2 (SSX2), Receptor for Advanced Glycation Endproducts (RAGE-1), renal ubiquitous 1 (RU1), renal ubiquitous 2 (RU2), legumain, human papilloma virus E6 (HPV E6), human papilloma virus E7 (HPV E7), intestinal carboxyl esterase, heat shock protein 70-2 mutated (mut hsp70-2), CD79a, CD79b, CD72, Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), Fc fragment of IgA receptor (FCAR or CD89), Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), CD300 molecule-like family member f (CD300LF), C-type lectin domain family 12 member A (CLEC12A), bone marrow stromal cell antigen 2 (BST2), EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2), lymphocyte antigen 75 (LY75), Glypican-3 (GPC3), Fc receptor-like 5 (FCRL5), FcRH5, PDL1, CD47, prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), Ron Kinase, c- Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin- B1, 9D7, Ep-CAM, EphA3, SAP-1, PRAME, Melan-A / MART-1, TRP1 / gp75, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, Ras, TGF-Β receptor, TGF-β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, gangliosides, MART-2, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, L1-CAM, gpA33, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, TGF-beta, hyaluronic acid, collagen, tenascin C, tenascin W, and immunoglobulin lambda-like polypeptide 1 (IGLL1).

62. The multifunctional molecule of any one of claims 59-61, wherein the tumor-targeting moiety is anantibody molecule.

63. The multifunctional molecule of claim 62, wherein the antibody molecule is selected from the groupconsisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

64. The multifunctional molecule of any one of claims 1-63, wherein the multifunctional moleculefurther comprises a stromal modifying moiety.

65. The multifunctional molecule of any one of claims 1-64, wherein the multifunctional moleculefurther comprises an immune cell engager.WSGR Docket No.53676-768.60166. The multifunctional molecule of claim 65, wherein the immune cell engager is selected from thegroup consisting of a T cell engager, an NK cell engager, a B cell engager, a dendritic cell engager, the macrophage cell engager, and any combination thereof.

67. The multifunctional molecule of claim 65 or 66, wherein immune cell engager is an antibodymolecule.

68. The multifunctional molecule of claim 67, wherein the antibody molecule is selected from the groupconsisting of a full-length antibody, an Fab, an Fab′, an F(ab′)2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a camelid antibody, a nanobody, and a single chain variable fragment (scFv).

69. The multifunctional molecule of any one of claims 1-68, wherein the first portion of the dimerizationmodule of the first polypeptide chain is a first Fc region or variant thereof.

70. The multifunctional molecule of any one of claims 1-69, wherein the second portion of thedimerization module of the second polypeptide chain is a second Fc region or variant thereof.

71. The multifunctional molecule of claim 69 or 70, wherein the first portion of the dimerization module,the second portion of the dimerization module, or a combination thereof is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ Fc region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

72. The multifunctional molecule of any one of claims 69-71, wherein the first Fc region, the second Fcregion, or a combination thereof comprises an engineered Fc interface with one or more of: a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, wherein dimerization of the first Fc region and the second Fc region is enhanced as indicated by a greater ratio of heteromultimer:homomultimer forms relative to dimerization of Fc regions without the engineered interface.

73. The multifunctional molecule of any one of claims 69-72, wherein the first Fc region, the second Fcregion, or a combination thereof comprises a Cys at position 349, a Ser at position 366, an Ala at position 368, a Val at position 407, a Cys at position 354, a Trp at position 366, or any combination thereof in a heavy chain constant region according to EU Numbering.

74. The multifunctional molecule of any one of claims 69-73, wherein the first Fc region, the second Fcregion, or a combination thereof comprises (i) a Cys at position 349, a Ser at position 366, an Ala at position 368, and a Val at position 407 in a heavy chain constant region according to EU Numbering; (ii) a Cys at position 354 and a Trp at position 366 in a heavy chain constant region according to EU Numbering; or (iii) a combination thereof.

75. The multifunctional molecule of any one of claims 69-74, wherein:(i) the first Fc region comprises:WSGR Docket No.53676-768.601 (a) a Cys at position 349 in a heavy chain constant region according to EU Numbering, (b) a Ser at position 366 in a heavy chain constant region according to EU Numbering, (c) an Ala at position 368 in a heavy chain constant region according to EU Numbering, and (d) a Val at position 407 in a heavy chain constant region according to EU Numbering; and (ii) the second Fc region comprises: (a) a Cys at position 354 in a heavy chain constant region according to EU Numbering, and (b) a Trp at position 366 in a heavy chain constant region according to EU Numbering.

76. The multifunctional molecule of any one of claims 69-74, wherein:(i) the first Fc region comprises: (a) a Cys at position 354 in a heavy chain constant region according to EU Numbering, and (b) a Trp at position 366 in a heavy chain constant region according to EU Numbering (ii) the second Fc region comprises: (a) a Cys at position 349 in a heavy chain constant region according to EU Numbering, (b) a Ser at position 366 in a heavy chain constant region according to EU Numbering, (c) an Ala at position 368 in a heavy chain constant region according to EU Numbering, and (d) a Val at position 407 in a heavy chain constant region according to EU Numbering.

77. The multifunctional molecule of any one of claims 69-76, wherein the first Fc region, the second Fcregion, or a combination thereof comprise one or more mutations that result in reduced or ablated affinity for at least one Fc receptor relative to a Fc region without the one or more mutations.

78. The multifunctional molecule of any one of claims 69-77, wherein the first Fc region, the second Fcregion, or a combination thereof comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

79. The multifunctional molecule of any one of claims 69-78, wherein the first Fc region, the second Fcregion, or a combination thereof comprises a sequence having at least 70% sequence identity to the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 3901, SEQ ID NO: 3645, SEQ ID NO: 3902, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO: 3903, SEQ ID NO:206, SEQ ID NO: 207, SEQ ID NO: 3904, SEQ ID NO: 3452, SEQ ID NO: 3447, or SEQ ID NO: 3453.

80. The multifunctional molecule of any one of claims 69-79, wherein the first Fc region, the second Fcregion, or a combination thereof comprises the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 3901, SEQ ID NO: 3645, SEQ ID NO: 3902, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO: 3903, SEQ ID NO:206, SEQ ID NO: 207, SEQ ID NO: 3904, SEQ ID NO: 3452, SEQ ID NO: 3447, or SEQ ID NO: 3453.

81. The multifunctional molecule of any one of claims 69-80, wherein the first Fc region, the second Fcregion, or a combination thereof comprises a sequence having at least 70% sequence identity to the sequence of SEQ ID NO:206, SEQ ID NO: 3447, or SEQ ID NO: 3453.

82. The molecule of any one of claims 69-81, wherein the first Fc region, the second Fc region, or acombination thereof comprises the sequence of SEQ ID NO:206, SEQ ID NO: 3447, or SEQ ID NO: 3453.WSGR Docket No.53676-768.60183. The multifunctional molecule of any one of claims 1-82, wherein the multifunctional the moleculecomprises the following configuration: A-[first portion of dimerization module]-C, B-[second portion of dimerization module]-D, wherein: (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first domain; (iii) the second domain, (iv) the at least one cytokine or a functional fragment or variant thereof, (v) the tumor targeting moiety, (vi) the stromal modifying moiety; or (vii) the immune cell engager; wherein at least one of A or C are the first domain, and wherein when A is the first domain, at least one of B, C, or D is the second domain, and when C is the first domain, at least one of A, B, or D is the second domain.

84. The multifunctional molecule of any one of claims 1-83, wherein the multifunctional molecule is notimmobilized to a solid-phase.

85. The multifunctional molecule of any one of claims 1-84, wherein the multifunctional molecule is notimmobilized to a solid-phase.

86. The multifunctional molecule any one of claim 1-85, wherein the second domain binds to one ormore of a TCRβV subfamily selected from the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 subfamily, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 subfamily, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

87. The multifunctional molecule of any one of claims 1-86, wherein the second domain binds to one ormore of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V1 subfamily comprising TCRβ V1*01;(ii) TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02,and TCRβ V2*03; (iii) TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; (iv) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04;WSGR Docket No.53676-768.601(v) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04, TCRβ V5-5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5-8*01, and TCRβ V5-8*02;(vi) TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6-3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6-6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6-9*01;(vii) TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7-3*03, TCRβ V7-3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7-7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7- 8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7-9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7-9*06, and TCRβ V7-9*07.(viii) TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02;(ix) TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-1*02, and TCRβ V9-1*03;(x) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10- 3*02, TCRβ V10-3*03, and TCRβ V10-3*04;(xi) TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02,TCRβ V11-2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11-3*03, and TCRβ V11- 3*04;(xii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-3*01, TCRβV12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01;(xiii) TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβV13*02;(xiv) TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβV14*02;(xv) TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβV15*02, and TCRβ V15*03;(xvi) TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβV16*02, and TCRβ V16*03;(xvii) TCRβ V17 subfamily comprising TCRβ V17*01;(xviii) TCRβ V18 subfamily comprising TCRβ V18*01;(xix) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβV19*02, and TCRβ V19*03;WSGR Docket No.53676-768.601 (xx) TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβV20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07; (xxi) TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβV21-1*02; (xxii) TCRβ V22 subfamily comprising TCRβ V22-1*01;(xxiii) TCRβ V23 subfamily comprising TCRβ V23-1*01;(xxiv) TCRβ V24 subfamily comprising TCRβ V24-1*01;(xxv) TCRβ V25 subfamily comprising TCRβ V25-1*01;(xxvi) TCRβ V26 subfamily comprising TCRβ V26-1*01;(xxvii) TCRβ V27 subfamily comprising TCRβ V27*01;(xxviii) TCRβ V28 subfamily comprising TCRβ V28*01;(xxix) TCRβ V29 subfamily comprising one or more selected from TCRβ V29-1*01, TCRβV29-1*02, and TCRβ V29-1*03; and (xxx) TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβV30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

88. The multifunctional molecule of any one of claims 1-87, wherein the second domain binds to TCRβV4 subfamily, TCRβ V6 subfamily, TCRβ V20 subfamily, or TCRβ V25 subfamily.

89. The multifunctional molecule of any one of claims 1-88, wherein the second domain binds to TCRΒV4-1, TCRΒ V6-1, TCRΒ V6-2, TCRΒ V6-3, TCRΒ V6-4, TCRΒ V6-5, TCRΒ V6-6, TCRΒ V6-7, TCRΒ V6-8, TCRΒ V6-9, TCRΒ V20-1, or TCRΒ V25-1.

90. The multifunctional molecule of any one of claims 1-89, wherein the multifunctional moleculecomprises a single TCRβV-binding domain.

91. The multifunctional molecule of any one of claims 1-90, wherein the first domain binds to one ormore of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

92. The multifunctional molecule of any one of claims 1-91, wherein the first domain binds to one ormore of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02,TCRα V1-2*01, and TCRα V1-2*02;WSGR Docket No.53676-768.601(ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02;(iii) TCRα V3 subfamily comprising TCRα V3*01;(iv) TCRα V4 subfamily comprising TCRα V4*01;(v) TCRα V5 subfamily comprising TCRα V5*01(vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02,TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06;(vii) TCRα V7 subfamily comprising TCRα V7*01;(viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8-4*01, TCRα V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01;(ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01,TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04;(x) TCRα V10 subfamily comprising TCRα V10*01;(xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02;(xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02;(xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRαV14*02, TCRα V14*03, and TCRα V14*04;(xiv) TCRα V16 subfamily comprising TCRα V16*01;(xv) TCRα V17 subfamily comprising TCRα V17*01;(xvi) TCRα V18 subfamily comprising TCRα V18*01;(xvii) TCRα V19 subfamily comprising TCRα V19*01;(xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRαV20*02, TCRα V20*03, and TCRα V20*04;(xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRαV20*02;(xx) TCRα V22 subfamily comprising TCRα V22*01;(xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRαV23*02, TCRα V23*03, and TCRα V23*04;(xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRαV23*02;(xxiii) TCRα V25 subfamily comprising TCRα V25*01;(xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRαV26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02;WSGR Docket No.53676-768.601 (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRαV27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRαV27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRαV30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01;(xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRαV35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRαV36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRαV38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01;(xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and(xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

93. The multifunctional molecule of any one of claims 1-92, wherein the second domain binds to TCRαV12 subfamily, TCRα V13 subfamily, TCRα V19 subfamily, TCRα V21 subfamily, or TCRα V30 subfamily.

94. The multifunctional molecule of any one of claims 1-92, wherein the second domain binds to TCRαV1 subfamily, TCRα V10 subfamily, TCRα V17 subfamily, or TCRα V19 subfamily.

95. The multifunctional molecule of any one of claims 1-92, wherein the second domain binds to TCRαV1-2, TCRα V10, or TCRα V17.

96. The multifunctional molecule of claim 1-95, wherein the first domain and the second domain bind to:TCRα V1 and TCRβ V6, respectively; TCRα V1 and TCRβ V20, respectively; TCRα V10 and TCRβ V25, respectively; TCRα V1 and TCRβ V4, respectively; TCRα V17 and TCRβ V4, respectively; or TCRα V17 and TCRβ V6, respectively.

97. The multifunctional molecule of claim 1-96, wherein the first domain and the second domain bind to:TCRα V1-2 and TCRβ V6-1, respectively; TCRα V1-2 and TCRβ V6-2, respectively; TCRα V1-2 and TCRβ V6-3, respectively; TCRα V1-2 and TCRβ V6-4, respectively; TCRα V1-2 and TCRβ V6-5, respectively; TCRα V1-2 and TCRβ V6-6, respectively; TCRα V1-2 and TCRβ V6-7, respectively;WSGR Docket No.53676-768.601 TCRα V1-2 and TCRβ V6-8, respectively; TCRα V1-2 and TCRβ V6-9, respectively; TCRα V1-2 and TCRβ V20-1, respectively; TCRα V10 and TCRβ V25-1, respectively; TCRα V1-2 and TCRβ V4-1, respectively; TCRα V17 and TCRβ V4-1, respectively; or TCRα V17 and TCRβ V6-2, respectively.

98. The multifunctional molecule of any one of claims 1-97, wherein the first domain comprises anantibody molecule that binds to the TCRαV comprising (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Table 22; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Table 22; or (iii) a combination thereof.

99. The multifunctional molecule of any one of claims 1-98, wherein the first domain comprises anantibody molecule that binds to the TCRαV comprising (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

100. The multifunctional molecule of any one of claims 1-99, wherein the first domain comprises anantibody molecule that binds to the TCRαV comprising (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Table 22; (i) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

101. The multifunctional molecule of any one of claims 1-100, wherein the first domain comprises anantibody molecule that binds to the TCRαV comprising (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

102. The multifunctional molecule of any one of claims 1-101, wherein the first domain comprises anyone of antibody molecules that binds to the TCRαV listed in Table 22.

103. The multifunctional molecule of any one of claims 1-102, wherein the second domain comprises anantibody molecule that binds to the TCRβV comprising (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

104. The multifunctional molecule of any one of claims 1-103, wherein the second domain comprises anantibody molecule that binds to the TCRβV comprising (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VL comprisingWSGR Docket No.53676-768.601 a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

105. The multifunctional molecule of any one of claims 1-104, wherein the second domain comprises anantibody molecule that binds to the TCRβV comprising (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (i) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

106. The multifunctional molecule of any one of claims 1-105, wherein the second domain comprises anantibody molecule that binds to the TCRβV comprising (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26; or (iii) a combination thereof.

107. The multifunctional molecule of any one of claims 1-106, wherein the second domain comprisesany one of antibody molecules that binds to the TCRβV listed in Tables 1, 2, 10A,10B, 10C, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 23, 25, or 26.

108. The multifunctional molecule of any one of claims 1-107, wherein the first Fc region, the second Fcregion, or a combination thereof comprises one or more mutations listed in tables 4 and 14 according to EU numbering.

109. The multifunctional molecule of any one of claims 1-108, wherein the first Fc region, the second Fcregion, or a combination thereof comprises a sequence having at least 70% sequence identity to any one of the heavy chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

110. The multifunctional molecule of any one of claims 1-109, wherein the first Fc region, the second Fcregion, or a combination thereof comprises any one of the heavy chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

111. The multifunctional molecule of any one of claims 1-110, wherein the multifunctional moleculefurther comprises one or more immunoglobulin light chain constant regions.

112. The multifunctional molecule of claim 111, wherein the one or more immunoglobulin light chainconstant region is linked to the first domain, the first portion of the first domain, or the second portion of the first domain.

113. The multifunctional molecule of claim 111 or 112, wherein the one or more immunoglobulin lightchain constant region is linked to the second domain, the first portion of the second domain, or the second portion of the second domain.WSGR Docket No.53676-768.601114. The multifunctional molecule of any one of claims 111-113, wherein the one or moreimmunoglobulin light chain constant region is linked to the at least one cytokine or a functional fragment or variant thereof.

115. The multifunctional molecule of any one of claims 111-114, wherein the one or moreimmunoglobulin light chain constant region comprises a kappa light chain constant region, a lambda light chain constant region, or a combination thereof.

116. The multifunctional molecule of any one of claims 111-115, wherein the one or moreimmunoglobulin light chain constant region comprises a sequence having at least 70% sequence identity to any one of the light chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

117. The multifunctional molecule of any one of claims 111-116, wherein the one or moreimmunoglobulin light chain constant region comprises any one of the light chain constant region sequences listed in Tables 1, 2, 3, 10C, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

118. The multifunctional molecule of any one of claims 33-117, wherein the at least one cytokine or afunctional fragment or variant thereof comprises a sequence having at least 70% sequence identity to any one of the cytokine sequences listed in Tables 10C, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

119. The multifunctional molecule of any one of claims 33-118, wherein the at least one cytokine or afunctional fragment or variant thereof comprises any one of the cytokine sequences listed in Tables 10C, 15, 16, 17, 18, 19, 20, 21, 22, 23, 25, or 26.

120. The multifunctional molecule of any one of claims 59-119, wherein the tumor-targeting moietycomprises a sequence having at least 70% sequence identity to any one of the antibody sequences listed in Table 24.

121. The multifunctional molecule of any one of claims 59-120, wherein the tumor-targeting moietycomprises any one of the antibody sequences listed in Table 24.

122. The multifunctional molecule of any one of claims 1-121, wherein the multifunctional moleculedoes not comprise an anti-CD3 binding domain.

123. The multifunctional molecule of any one of claims 1-122, wherein the multifunctional molecule is apolypeptide molecule.

124. The multifunctional molecule of any one of claims 1-123, wherein the multifunctional molecule is amultispecific molecule.

125. A polynucleotide comprising a sequence encoding the multifunctional molecule of any one ofclaims 1-124.

126. The polynucleotide of claim 125, wherein the polynucleotide is an isolated nucleic acid molecule.

127. A vector comprising the polynucleotide of claim 125 or 126.

128. A cell comprising the multifunctional molecule of any one of claims 1-124, the polynucleotide ofclaim 125 or 126, or the vector of claim 127.WSGR Docket No.53676-768.601129. A method of making the multifunctional molecule of any one of claims 1-124 comprising: culturinga cell comprising the polynucleotide of claim 125 or 126 or the vector of claim 127 under conditions suitable for expression of the multifunctional molecule.

130. A method of making the multifunctional molecule of any one of claims 1-124 comprising: culturingthe cell of claim 128 under conditions suitable for expression of the multifunctional molecule.

131. A composition comprising the multifunctional molecule of any one of claims 1-124.

132. A pharmaceutical composition comprising the multifunctional molecule of any one of claims 1-124,the polynucleotide of claim 125 or 126, the vector of claim 127, the cell of claim 128, or the composition of claim 131, and a pharmaceutically acceptable carrier, excipient, or diluent.

133. A method of treating a condition or disease in a subject in need thereof comprising administering tothe subject a therapeutically effective amount of the multifunctional molecule of any one of claims 1- 124, the polynucleotide of claim 125 or 126, the vector of claim 127, the cell of claim 128, the composition of claim 131, the pharmaceutical composition of claim 132, or a combination thereof, wherein the administering is effective to treat the condition or disease in the subject.

134. The method of claim 133, wherein the condition or disease is cancer.

135. The method of claim 134, wherein the cancer is a solid tumor, a hematological cancer, a metastaticcancer, a soft tissue tumor, or a combination thereof.

136. The method of claim 134 or 135, wherein the cancer is the solid tumor, and wherein the solid tumoris selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

137. The method of claim 134 or 135, wherein the cancer is the hematological cancer, and wherein thehematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T-cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

138. The method of claim 137, wherein the Non-Hodgkin’s lymphoma is selected from the groupconsisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

139. The method of claim 137, wherein the T-cell lymphoma is peripheral T-cell lymphoma.

140. The method of any one of claims 134-139, wherein the cancer is characterized by a cancer antigenpresent on the cancer.

141. The method of claim 140, wherein the cancer antigen present on the cancer is a tumor antigen, astromal antigen, or a hematological antigen.

142. The method of claim 140 or 141, wherein the cancer antigen is selected from the group consisting ofCD19, CD123, CD22, CD30, CD171, CS-1, C-type lectin-like molecule-1, CD33, epidermal growth factor receptor variant III (EGFRvIII), ganglioside G2 (GD2), ganglioside GD3, TNF receptor family member B cell maturation (BCMA), Tn antigen ((Tn Ag) or (GalNAcα-Ser / Thr)), prostate-specific membrane antigen (PSMA), Receptor tyrosine kinase-like orphan receptor 1 (ROR1), Fms-LikeWSGR Docket No.53676-768.601 Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD117), Interleukin-13 receptor subunit alpha-2, mesothelin, Interleukin 11 receptor alpha (IL- 11Ra), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage-specific embryonic antigen-4 (SSEA-4), CD20, Folate receptor alpha, Receptor tyrosine-protein kinase ERBB2 (Her2 / neu), Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion molecule (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gp100) / pmel17, oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), tyrosinase, ephrin type-A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma-associated antigen (HMWMAA), o- acetyl-GD2 ganglioside (OAcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7-related (TEM7R), claudin 6 (CLDN6), thyroid stimulating hormone receptor (TSHR), G protein-coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, CD179a, anaplastic lymphoma kinase (ALK), Polysialic acid, placenta-specific 1 (PLAC1), hexasaccharide portion of globoH glycoceramide (GloboH), mammary gland differentiation antigen (NY-BR-1), uroplakin 2 (UPK2), Hepatitis A virus cellular receptor 1 (HAVCR1), adrenoceptor beta 3 (ADRB3), pannexin 3 (PANX3), G protein-coupled receptor 20 (GPR20), lymphocyte antigen 6 complex, locus K 9 (LY6K), Olfactory receptor 51E2 (OR51E2), TCR Gamma Alternate Reading Frame Protein (TARP), Wilms tumor protein (WT1), Cancer / testis antigen 1 (NY-ESO-1) / LAGE-1, Cancer / testis antigen 2 (LAGE-1a), Melanoma-associated antigen 1 (MAGE-A1), ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML), sperm protein 17 (SPA17), X Antigen Family, Member 1A (XAGE1), angiopoietin-binding cell surface receptor 2 (Tie 2), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), Fos-related antigen 1, tumor protein p53 (p53), p53 mutant, prostein, survivin, telomerase, prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1, Rat sarcoma (Ras) mutant, human Telomerase reverse transcriptase (hTERT), sarcoma translocation breakpoints, melanoma inhibitor of apoptosis (ML-IAP), ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene), N-Acetyl glucosaminyl-transferase V (NA17), paired box protein Pax-3 (PAX3), Androgen receptor, Cyclin B1, v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN), Ras Homolog Family Member C (RhoC), Tyrosinase-related protein 2 (TRP-2), Cytochrome P4501B1 (CYP1B1), CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3), Paired box protein Pax-5 (PAX5), proacrosin binding proteinWSGR Docket No.53676-768.601 sp32 (OY-TES1), lymphocyte-specific protein tyrosine kinase (LCK), A kinase anchor protein 4 (AKAP-4), synovial sarcoma, X breakpoint 2 (SSX2), Receptor for Advanced Glycation Endproducts (RAGE-1), renal ubiquitous 1 (RU1), renal ubiquitous 2 (RU2), legumain, human papilloma virus E6 (HPV E6), human papilloma virus E7 (HPV E7), intestinal carboxyl esterase, heat shock protein 70-2 mutated (mut hsp70-2), CD79a, CD79b, CD72, Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), Fc fragment of IgA receptor (FCAR or CD89), Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), CD300 molecule-like family member f (CD300LF), C-type lectin domain family 12 member A (CLEC12A), bone marrow stromal cell antigen 2 (BST2), EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2), lymphocyte antigen 75 (LY75), Glypican-3 (GPC3), Fc receptor-like 5 (FCRL5), FcRH5, PDL1, CD47, prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), Ron Kinase, c- Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin- B1, 9D7, Ep-CAM, EphA3, SAP-1, PRAME, Melan-A / MART-1, TRP1 / gp75, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, Ras, TGF-Β receptor, TGF-β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, gangliosides, MART-2, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, L1-CAM, gpA33, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, TGF-beta, hyaluronic acid, collagen, tenascin C, tenascin W, and immunoglobulin lambda-like polypeptide 1 (IGLL1).

143. The method of any one of claims 134-142, further comprising administering a second therapeuticagent or therapy to the subject.

144. The method of claim 143, wherein the second therapeutic agent or therapy comprises achemotherapeutic agent, a biologic agent, a hormonal therapy, radiation, or surgery.

145. The method of claim 143 or 144, wherein the second therapeutic agent or therapy is administered incombination with the multifunctional molecule of any one of claims 1-124, the polynucleotide of claim 125 or 126, the vector of claim 127, the cell of claim 128, , the composition of claim 131, or the pharmaceutical composition of claim 132, sequentially, simultaneously, or concurrently.

146. A method of expansion of a subset of T cells in a T cell population comprising contacting the T cellpopulation with the multifunctional molecule of any one of claims 1-124 or the composition of claim 131, thereby expanding the subset of T cells in the T cell population.

147. The method of claim 146, wherein the T cell population is a human T cell population.

148. The method of claim 146 or 147, wherein the subset of T cells are a subset of human T cells.

149. The method of any one of claims 146-148, wherein the subset of T cells express a T cell receptor(TCR) comprising a TCRβ chain that comprises the TCRβV to which the second domain binds.

150. The method of any one of claims 146-149, wherein the subset of T cells express a TCR comprisinga TCRα chain that comprises the TCRαV to which the first domain binds.

151. The method of claim 149 or 150, wherein the TCR is a human TCR.WSGR Docket No.53676-768.601152. The method of any one of claims 149-151, wherein the multispecific molecule is an agonist of theTCR.

153. The method of any one of claims 146-152, wherein the T cell population is an in vivo T cellpopulation.

154. The method of any one of claims 146-152, wherein the T cell population is an ex vivo T cellpopulation.

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