CD161 binding proteins and related methods

Abstract

Provided herein are CD161 binding proteins (e.g., antibodies) and compositions (e.g., pharmaceutical compositions) comprising the same; as well as methods of making the CD161 binding proteins (e.g., antibodies) and compositions. Further provided herein are kits comprising the CD161 binding proteins (e.g., antibodies) described herein. The CD161 binding proteins provided herein are useful in e.g., in therapeutic and diagnostic methods.

Description

Attorney Docket No. 63340.32WO01CD161 BINDING PROTEINS AND RELATED METHODS RELATED APPLICATIONS

[0001] This application claims priority to U.S. Serial No.: 63 / 721,878, filed November 18, 2024, U.S. Serial No.: 63 / 733,609, filed December 13, 2024, and U.S. Serial No.: 63 / 831,646, filed June 27, 2025, the entire contents of each of which is incorporated herein by reference.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on November 15, 2025, is named 63340_32WO01_SL.xml and is 65,870 bytes in size.1. FIELD

[0003] This disclosure relates to CD 161 binding proteins and nucleic acid molecules encoding the same. This disclosure is further related to methods of manufacturing and utilizing the same, including e.g., methods of treatment of a disease (e.g., a proinflammatory disease) in a subject in need thereof.2. BACKGROUND

[0004] CD 161 is a C-type lectin-like type-II transmembrane protein. CD 161 is encoded by the killer cell lectin like receptor Bl (KLRB1) gene located within the natural killer (NK) cell gene complex on chromosome 12. CD161 is expressed by e.g., NK cells and subsets of both CD4+ and CD8+ T cells. CD161 binds CLEC2D, also a C-type lectin transmembrane protein. CLEC2D is expressed, e.g., on the surface of both malignant cells and immune cells including germinal center B cells, activated T cells, and tumor associated macrophages. CLEC2D / CD161 interactions play a role in regulating immune responses in various contexts.3. SUMMARY

[0005] Provided herein are, inter alia, CD 161 binding proteins and nucleic acid molecules encoding the same; fusions and conjugates comprising the CD161 binding proteins; methods of manufacturing; pharmaceutical compositions; and methods of use including e.g., methods ofAttorney Docket No. 63340.32WO01treating a disease (e.g., a proinfl ammatory disease) in a subject in need thereof, methods of inhibiting binding of CD 161 to CLEC2D, and diagnostics.

[0006] Accordingly, in one aspect, provided herein are CD 161 binding proteins comprising a binding domain that specifically binds CD161 (e.g., human CD161 (hCD161)), wherein the binding domain comprises: (a) a VH region comprising (i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 3 or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid modifications; (ii) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 4 or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid modifications; and (iii) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 5 or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid modifications; and (b) a VL region comprising (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 6 or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid modifications; (ii) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 7 or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1. 2, or 3 amino acid modifications; and (iii) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 8 or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid modifications.

[0007] In one aspect, provided herein are CD 161 binding protein comprising a binding domain that specifically binds CD161 (e.g., hCD161), wherein the binding domain comprises: (a) a VH region comprising (i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 3; (ii) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 4; and (iii) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 5: and (b) a VL region comprising (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 6; (ii) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 7; and (iii) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 8.

[0008] In one aspect, provided herein are CD 161 binding protein comprising a binding domain that specifically binds CD161 e.g., hCD161), wherein the binding domain comprises: (a) a VH region comprising an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and (b) a VL region comprising an amino acid sequence at least 85%, 86%. 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to theAttorney Docket No. 63340.32WO01amino acid sequence set forth in SEQ ID NO: 10.

[0009] For the sake of clarity, it should be understood that the following embodiments are applicable to each of the foregoing aspects as if recited directly following each individual aspect.

[0010] In some embodiments, (a) the amino acid sequence of the VH region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and (b) the amino acid sequence of the VL region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

[0011] In some embodiments, (a) the amino acid sequence of the VH region is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and (b) the amino acid sequence of the VL region is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

[0012] In some embodiments, (a) the amino acid sequence of the VH region is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and (b) the amino acid sequence of the VL region is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

[0013] In some embodiments, (a) the amino acid sequence of the VH region is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 9; and (b) the amino acid sequence of the VL region is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 10.

[0014] In some embodiments, (a) the amino acid sequence of the VH region comprises the amino acid sequence set forth in SEQ ID NO: 9; and (b) the amino acid sequence of the VL region comprises the amino acid sequence set forth in SEQ ID NO: 10.

[0015] In some embodiments, the CD161 binding protein comprises a heavy chain (HC). In some embodiments, the CD 161 binding protein comprises a light chain (LC).

[0016] In some embodiments, the CD 161 binding protein comprises a heavy chain (HC) and a light chain (LC).

[0017] In some embodiments, the CD 161 binding protein comprises the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,Attorney Docket No. 63340.32WO0195%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68.

[0018] In some embodiments, the CD 161 binding protein (or the binding domain) comprises an antibody.

[0019] In some embodiments, the CD 161 binding protein (or the binding domain) comprises an IgG (e.g., IgGi, IgGz (e.g., lgG2;, or IgG2b), IgGs. IgG4), IgE, IgM, IgD, or IgA (e.g., IgAi or IgA2) antibody.

[0020] In some embodiments, the CD 161 binding protein (or the binding domain) comprises one or more of a monoclonal antibody, monospecific antibody, multispecific antibody, human antibody, humanized antibody, chimeric antibody, and / or murine antibody, or a functional fragment or functional variant of any of the foregoing.

[0021] In some embodiments, the CD 161 binding protein (or the binding domain) comprises one or more of a full-length antibody, scFv, Fab, F(ab’)2, Fab', Fv, single domain antibody (e.g., a VHH), scFv-Fc, Fab-Fc, and / or single domain antibody-Fc (e.g., VHH-Fc).

[0022] In some embodiments, the CD 161 binding protein (e.g., an anti-CD161 antibody) comprises a hlg Fc region.

[0023] In some embodiments, the hlg Fc region is capable of mediating one or more hlg Fc effector function. In some embodiments, the hlg Fc region is capable of mediating one or more of ADCC, ADCP, or CDC. In some embodiments, the hlg Fc region is capable of mediating ADCC.

[0024] In some embodiments, the hlg Fc region comprises one or more amino acid variation or alteration (e.g., described herein (e.g., Table 4 herein)) in glycosylation that enhances one or more hlg Fc effector function (e.g., compared to a reference hlg Fc region or a reference CD161 binding protein comprising the same).

[0025] In some embodiments, the hlg Fc region comprises one or more amino acid variation or alteration in glycosylation that enhances ADCC and / or ADCP (e.g., compared to a reference hlg Fc region or a reference CD161 binding protein comprising the same).

[0026] In some embodiments, the hlg Fc region is afucosylated.

[0027] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) is capable of inhibiting (blocking) the binding of CD 161 to CEEC2D.

[0028] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) is capable of mediating depletion of a population of CD161 expressing cells (e.g., immune cells (e.g.,Attorney Docket No. 63340.32WO01T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) (e.g., upon binding to CD161 expressed on the surface of the population of cells).

[0029] In some embodiments, the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells), NK cells, MAIT cells, gd T cells, Tregs, Th2A cells, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more pro-inflammatory cytokine is IL-4, IL-13, IL-5 or IL-9. In some embodiments, the population of CD161 expressing cells comprises Th2A cells and the one or more pro-inflammatory cytokine is IL-4, IL-13. IL-5 or IL-9.

[0030] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) is capable of mediating a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF) (e.g., produced by a population of cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs))). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0031] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector function.

[0032] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of antibody dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), or complement dependent cytotoxicity (CDC).

[0033] In some embodiments, the CD 161 binding protein comprises a heterologous moiety (e.g., a heterologous protein). In some embodiments, the CD161 binding protein comprises 2, 3, 4, or 5 or more heterologous moieties.

[0034] In some embodiments, the heterologous moiety is attached to the N-terminus, C-terminus, and / or internally between the N- and C-terminus of the CD161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologous polypeptide) is directly attached to the CD161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologousAttorney Docket No. 63340.32WO01polypeptide) is indirectly attached to the CD161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD161 binding protein via a linker.

[0035] In some embodiments, the heterologous moiety is a peptide, protein (e.g., antibody), nucleic acid molecule (e.g., DNA, RNA), carbohydrate, lipid, polymer, or small molecule. In some embodiments, the heterologous moiety is a half-life extension moiety (e.g., half-life extension protein (e.g., Fc region)). In some embodiments, the heterologous moiety is a detectable tag and / or a reporter gene. In some embodiments, the heterologous moiety is a therapeutic agent.

[0036] In some embodiments, the CD 161 binding protein is isolated. In some embodiments, the CD 161 binding protein is recombinant.

[0037] In one aspect, provided herein are conjugated comprising a CD 161 binding protein described herein and a heterologous moiety. In some embodiments, the conjugate comprises 2, 3, 4, or 5 or more heterologous moieties.

[0038] In some embodiments, the heterologous moiety is attached to the N-terminus, C-terminus, and / or internally between the N- and C-terminus of the CD161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologous polypeptide) is directly attached to the CD 161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD161 binding protein. In some embodiments, the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD161 binding protein via a linker.

[0039] In some embodiments, the heterologous moiety is a peptide, protein, carbohydrate, lipid, polymer, or small molecule. In some embodiments, the heterologous moiety is a therapeutic agent. In some embodiments, the therapeutic agent is a small molecule.

[0040] In one aspect, provided herein are fusion proteins comprising a CD 161 binding protein described herein and a heterologous protein.

[0041] In some embodiments, the heterologous protein comprises a cytokine (or functional fragment or functional variant thereof), a chemokine (or a functional fragment or functional variant thereof), or an antibody (or a functional fragment or functional variant thereof).

[0042] In some embodiments, the heterologous protein is fused to the N-terminus, C-terminus, and / or internally between the N- and C-terminus of the CD161 binding protein. In some embodiments, the heterologous protein is fused directly to the CD161 binding protein. In someAttorney Docket No. 63340.32WO01embodiments, the heterologous protein is fused indirectly to the CD161 binding protein. In some embodiments, the heterologous protein is fused indirectly to the CD 161 binding protein via a peptide linker.

[0043] In one aspect, provided herein are nucleic acid molecule encoding a CD 161 binding protein described herein or a fusion protein described herein.

[0044] In some embodiments, the nucleic acid molecule is a DNA or RNA (e.g., mRNA) molecule. In some embodiments, the nucleic acid molecule is codon optimized. In some embodiments, the nucleic acid molecule comprises one or more transcription or translation regulatory elements (e.g., promoter, enhancer (e.g., cell or tissue specific transcription regulatory elements).

[0045] In one aspect, provided herein are vectors comprising a nucleic acid molecule described herein. In some embodiments, the vector is a viral vector or a non-viral vector (e.g., plasmid, minicircle). In some embodiments, the carrier is a viral vector (e.g., an adeno associated viral (AAV) vector, a lentiviral vector, an adenoviral vector).

[0046] In one aspect, provided herein are carriers comprising a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a cell (or population of cells) described herein, or a pharmaceutical composition described herein. In some embodiments, the carrier is a nanoparticle, polymer, virus (e.g., a recombinant virus), virus like particle, virosome, fusosome, vesicle, or lipid-based carrier (e.g., a lipid nanoparticle (LNP), liposome, lipoplex, nanoliposome, an exosome, or a micelle).

[0047] In one aspect, provided herein are cells or population of cells comprising a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein.

[0048] In one aspect, provided herein are pharmaceutical compositions comprising a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, or a cell or population of cells described herein, and a pharmaceutically acceptable excipient.

[0049] In one aspect, provided herein are kits comprising a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid moleculeAttorney Docket No. 63340.32WO01described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein; and optionally instructions for using any one or more of the foregoing.

[0050] In one aspect, provided herein are methods of manufacturing a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, the method comprising: introducing into a cell a nucleic acid molecule described herein, a vector described herein, or a carrier described herein; culturing the cell under conditions that allow for expression of the CD161 binding protein, the conjugate, or the fusion protein; and optionally recovering the expressed the CD 161 binding protein, the conjugate, or the fusion protein from the culture; and optionally purifying the expressed the CD 161 binding protein, the conjugate, or the fusion protein from the culture.

[0051] In one aspect, provided herein are methods of manufacturing an afucosylated CD161 binding protein described herein, a fusion protein described herein, a conjugate described herein, the method comprising: introducing into a cell that comprises a disruption in the FUT8 gene a nucleic acid molecule described herein, a vector described herein, or a carrier described herein; culturing the cell under conditions that allow for expression of the afucosylated CD 161 binding protein, the conjugate, or the fusion protein; and optionally recovering the expressed the CD161 binding protein, the conjugate, or the fusion protein from the culture; and optionally purifying the expressed the CD 161 binding protein, the conjugate, or the fusion protein from the culture.

[0052] In one aspect, provided herein are methods of delivering a CD161 binding protein described herein, a conjugate described herein, a fusion protein described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell (or population of cells) described herein, or a pharmaceutical composition described herein to a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby deliver the CD 161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or pharmaceutical composition to the subject.

[0053] In one aspect, provided herein are methods of treating a proinflammatory disease (e.g., an autoimmune disease) in a subject in need thereof, the method comprising administering to theAttorney Docket No. 63340.32WO01subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby treat the proinflammatory disease (e.g., the autoimmune disease) in the subject.

[0054] In some embodiments, the proinflammatory disease (e.g., the autoimmune disease) is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis. In some embodiments, the proinflammatory disease is an autoimmune disease, an idiopathic inflammatory myopathies (e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

[0055] In some embodiments, the proinflammatory disease is an allergic disease (e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder (e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgE mediated disease (e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

[0056] In some embodiments, the autoimmune disease is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, an idiopathic inflammatory myopathies (e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), or sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

[0057] In some embodiments, the autoimmune disease is an allergic disease (e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder (e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgEAttorney Docket No. 63340.32WO01mediated disease (e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

[0058] In some embodiments, the proinflammatory disease (e.g., an autoimmune disease) is treated, at least in part, through the depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells. MAIT cells, gd T cells. Tregs. and / or ILCs)) in the subject.

[0059] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) mediates depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) in the subject.

[0060] In some embodiments, the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0061] In some embodiments, the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0062] In some embodiments, the proinflammatory disease (e.g., an autoimmune disease) is treated, at least in part, through a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0063] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-Attorney Docket No. 63340.32WO0117, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinfl ammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more prointlammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0064] In some embodiments, the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF) is mediated by the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL-13, IL-5 or IL-9.

[0065] In some embodiments, the population of CD161 expressing cells comprises one or more subpopulations of immune cells.

[0066] In some embodiments, the population of CD 161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).

[0067] In some embodiments, the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells).

[0068] In some embodiments, the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

[0069] In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL- 5 or IL-9.

[0070] In one aspect, provided herein are methods of treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby treat the autoimmune disease in the subject.

[0071] In some embodiments, the autoimmune disease is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemicAttorney Docket No. 63340.32WO01lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, an idiopathic inflammatory myopathies {e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), or sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

[0072] In some embodiments, the autoimmune disease is an allergic disease {e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder {e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgE mediated disease {e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

[0073] In some embodiments, the autoimmune disease is treated, at least in part, through the depletion of a population of CD161 expressing cells {e.g., immune cells e.g., T cells {e.g., activated T cells) {e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) in the subject.

[0074] In some embodiments, the CD161 binding protein {e.g., the anti-CD161 antibody) mediates depletion of a population of CD161 expressing cells {e.g., immune cells {e.g., T cells {e.g., activated T cells) {e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) in the subject.

[0075] In some embodiments, the depletion of the population of CD161 expressing cells {e.g., immune cells {e.g., T cells {e.g., activated T cells) {e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine {e.g., IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IE-17, IE-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IE-4, IE- 13, IE-5 or IL-9.

[0076] In some embodiments, the autoimmune disease is treated, at least in part, through a reduction in the level of one or more proinflammatory cytokine {e.g., IFNy, TNFa, IE- 17, IE-22, IE-21, or GM-CSF). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL-13, IL-5 or IL-9.

[0077] In some embodiments, the CD161 binding protein {e.g., the anti-CD161 antibody) mediates a reduction in the level of one or more proinflammatory cytokine {e.g., IFNy, TNFa, IL-Attorney Docket No. 63340.32WO0117, IL-22, IL-21, or GM-CSF). In some embodiments, the one or more proinfl ammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more prointlammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0078] In some embodiments, the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF) is mediated by the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 or IL-9.

[0079] In some embodiments, the population of CD161 expressing cells comprises one or more subpopulations of immune cells.

[0080] In some embodiments, the population of CD 161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).

[0081] In some embodiments, the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells).

[0082] In some embodiments, the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

[0083] In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL- 5 or IL-9.

[0084] In one aspect, provided herein are methods of depleting a population of cells (e.g., immune cells (e.g., T cells, NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) expressing CD161 (e.g., on the surface) in a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby deplete the population of cells (e.g., immune cells (e.g., T cells, NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) expressing CD161 (e.g., on the surface) in the subject.

[0085] In some embodiments, the population of cells comprises T cells (e.g., activated T cells)Attorney Docket No. 63340.32WO01(e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs.

[0086] In some embodiments, the population of cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells).

[0087] In some embodiments, the population of cells comprises CD4+ T cells and CD8+ T cells (e.g., activated CD4+ T cells and CD8+ T cells).

[0088] In some embodiments, the population of cells expresses one or more proinflammatory cytokine.

[0089] In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL- 5 or IL-9.

[0090] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a human Ig (hlg) Fc region.

[0091] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector function.

[0092] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of antibody dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), or complement dependent cytotoxicity (CDC).

[0093] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells, at least in part, through ADCC.

[0094] In one aspect, provided herein are methods of reducing the level of one or more proinflammatory cytokines in a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby reduce the level of one or more proinflammatory cytokines in the subject.

[0095] In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL- 13, IL-5 and IL-9.Attorney Docket No. 63340.32WO01

[0096] In some embodiments, the level of a plurality of proinflammatory cytokines is reduced (e.g., simultaneously).

[0097] In some embodiments, the level of at least 2, 3, 4, 5, or 6, or more proinflammatory cytokines is reduced (e.g., simultaneously).

[0098] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a human Ig (hlg) Fc region.

[0099] In some embodiments, the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, or GM-CSF) is mediated by the depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL-13, IL-5 and IL-9.

[0100] In some embodiments, the population of CD161 expressing cells comprises one or more subpopulations of immune cells.

[0101] In some embodiments, the population of CD 161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).

[0102] In some embodiments, the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells).

[0103] In some embodiments, the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

[0104] In some embodiments, the CD 161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector function.

[0105] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of ADCC. ADCP, or CDC.

[0106] In some embodiments, the CD 161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells, at least in part, through ADCC.

[0107] In one aspect, provided herein are methods of inhibiting (blocking) binding of CD161 to CLEC2D expressed on the surface of a cell in a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector describedAttorney Docket No. 63340.32WO01herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby inhibit (blocking) binding of CD 161 to CLEC2D expressed on the surface of a cell in the subject.

[0108] In one aspect, provided herein are methods of treating cancer in a subject in need thereof, the method comprising administering to the subject a CD 161 binding protein described herein, a fusion protein described herein, a conjugate described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell or population of cells described herein, or a pharmaceutical composition described herein, to thereby treat the cancer in the subject.

[0109] In some embodiments, the cancer is a leukemia or lymphoma. In some embodiments, the cancer is a T cell leukemia, NK cell leukemia. T cell prolymphocytic leukemia (T-PLL), or large granular lymphocytic leukemia (LGLL). In some embodiments, the cancer is a NK / T cell lymphoma (NKTCL), extranodal NK / T cell lymphoma (ENKL), mycosis fungoides (MF), Sezary syndrome, peripheral T cell lymphoma, angioimmunoblastic T cell lymphoma (AITL), and peripheral T cell lymphoma not otherwise specified (PTCL-NOS).

[0110] In some embodiments, a CD161 binding protein described herein or a fusion protein described herein is administered to the subject.

[0111] In some embodiments, the CD161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or a pharmaceutical composition is administered in combination with one or more additional therapeutic agent. In some embodiments, the CD 161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or a pharmaceutical composition is administered prior to. concomitant with, and / or after the administration of one or more additional therapeutic agent. In some embodiments, the one or more additional therapeutic agent is an anti-inflammatory agent.

[0112] In some embodiments, the subject is human.4. BRIEF DESCRIPTION OF THE DRAWINGS

[0113] FIG. 1 is a line graph showing the binding of Ab-1 (SEQ ID NOS: 67-68) (at the indicated concentration) to CD 161 expressed on the surface of HEK cells (as measured by How cytometry). Binding expressed as mean fluorescence intensity.

[0114] FIG. 2 is a line graph showing the binding of Ab-1 (SEQ ID NOS: 67-68) (at theAttorney Docket No. 63340.32WO01indicated concentration) to human CD161 expressed on the surface of CD4+ T cells.

[0115] FIG. 3 is a line graph showing the binding of Ab-1 (SEQ ID NOS: 67-68) (at the indicated concentration) to cyno CD161 expressed on the surface of CD4+ T cells.

[0116] FIG. 4 is a line graph showing the binding of CD 161 expressed on the surface of cells in vitro to biotinylated CLEC2D-Fc fusion in presence of Ab-1 (at the indicated concentration) (or no antibody control).

[0117] FIG. 5 is a line graph showing Ab-1 induced ADCC mediated killing of CD 161 expressing Jurkat cells (at the indicated concentration of Ab-1).

[0118] FIG. 6 is a line graph showing Ab-1 induced ADCC mediated killing of CD161 expressing human T cells (at the indicated concentration of Ab-1).

[0119] FIG. 7 is a line graph showing Ab-1 and afucosylated Ab-1 induced ADCC mediated killing of CD161 expressing CD4+ T cells (at the indicated concentration of Ab-1).

[0120] FIG. 8A is a line graph showing the total PASI (Psoriasis Area and Severity index) score of cynomolgus monkeys treated with 10 mg / kg afucosylated Ab-1 (or vehicle control) in the IMQ mediated Psoriasis model. FIG. 8B is a line graph showing the total PASI (Psoriasis Area and Severity index) score of cynomolgus monkeys treated with 20 mg / kg afucosylated Ab-1 (or vehicle control) in the IMQ mediated Psoriasis model.

[0121] FIG. 9 is a line graph showing the total PASI (Psoriasis Area and Severity index) score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg) or a reference anti-IL-17 antibody (3 mg / kg) in the IMQ mediated Psoriasis model.

[0122] FIG. 10A is a line graph showing the erythema score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control in the IMQ mediated Psoriasis model. FIG.10B is a line graph showing the scaling score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control in the IMQ mediated Psoriasis model. FIG.10C is a line graph showing the thickening score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control in the IMQ mediated Psoriasis model. FIG. 10D is a line graph showing the total PASI score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control in the IMQ mediated Psoriasis model.Attorney Docket No. 63340.32WO01

[0123] FIG. 11 shows photographic images of the skin inflammation of cynomolgus monkeys treated with afucosylated Ab- 1 (10 mg / kg or 20 mg / kg), a reference anti-IL- 17 antibody (3 mg / kg), or a vehicle control in the IMQ mediated Psoriasis model. Representative images at Day 14.

[0124] FIG. 12 is a line graph showing the total PASI (Psoriasis Area and Severity index) score of cynomolgus monkeys treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg) or a reference anti-IL- 17 antibody (3 mg / kg) in the IMQ mediated Psoriasis model through day 29 of treatment.

[0125] FIG. 13A is a line graph showing the total PASI score for individual cynomolgus monkeys in the IMQ mediated Psoriasis model treated with vehicle control through day 29. FIG.13B is a line graph showing the total PASI score for individual cynomolgus monkeys in the IMQ mediated Psoriasis model treated with a reference anti-IL- 17 antibody (3 mg / kg) through day 29.FIG. 13C is a line graph showing the total PASI score for individual cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg) through day 29. FIG.13D is a line graph showing the total PASI score for individual cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) through day 29.

[0126] FIG. 14A is a line graph showing the average skin erythema of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL- 17 antibody (3 mg / kg), or a vehicle control through day 29. FIG. 14B is a line graph showing the average skin thickening of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL- 17 antibody (3 mg / kg), or a vehicle control through day 29. FIG. 14C is a line graph showing the average skin scaling of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control through day 29. FIG. 14D is a line graph showing the average total PASI score of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL- 17 antibody (3 mg / kg), or a vehicle control through day 29.

[0127] FIG. 15A is a bar graph showing the average total PASI score of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL- 17 antibody (3 mg / kg), or a vehicle control as area under the curve (AUC) measurements. FIG. 15B is a bar graph showing the average skin thickening of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20Attorney Docket No. 63340.32WO01mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control as area under the curve (AUC) measurements. FIG. 15C is a bar graph showing the average skin scaling of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control as area under the curve (AUC) measurements. FIG. 15D is a bar graph showing the average skin erythema of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control as area under the curve (AUC) measurements.

[0128] FIG. 16A is a FACS plot showing the number of CD161 expressing T cells from the peripheral blood of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) or a vehicle control. FIG. 16B is a line graph showing the percent of CD161 expressing T cells from the peripheral blood of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) (expressed as percent of total CD3+ cells) over 14 days.

[0129] FIG. 17A is dot plot showing the frequency of CD161 expressing CD8+ T cells in intestinal biopsies of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) or vehicle control at day 17. FIG. 17B is dot plot showing the frequency of CD161 expressing CD4+ T cells in intestinal biopsies of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) or vehicle control at day 17. FIG. 17C is dot plot showing the frequency of CD161 expressing NK cells in intestinal biopsies of cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (20 mg / kg) or vehicle control at day 17.

[0130] FIG. 18A is a bar graph showing the average histopathology score of skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control at day 0, day 7, or day 14. FIG. 18B is a bar graph showing the average lymphocyte infiltrate score of skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab- 1 (10 mg / kg or 20 mg / kg), a reference anti-IL-17 antibody (3 mg / kg), or a vehicle control at day 0, day 7, or day 14.

[0131] FIG. 19 presents microscopy images of hematoxylin and eosin stained skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1Attorney Docket No. 63340.32WO01(10 mg / kg) or a vehicle control at day 0 or day 14.

[0132] FIG. 20A is a heat map showing the change in expression level of the indicated pro-inflammatory cytokines in skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg) or vehicle control at day 15. FIG.20B is a heat map showing the change in expression level of the indicated chemokines in skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg) or vehicle control at day 15. FIG. 20C is a heat map showing the change in expression level of the indicated psoriatic skin lesion associated genes in skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model treated with afucosylated Ab-1 (10 mg / kg) or vehicle control at day 15.

[0133] FIG.21 is a line graph showing afucosylated Ab-1 induced ADCC mediated killing of CD 161 expressing cyno CD8+ T cells (at the indicated concentration of Ab-1).

[0134] FIG. 22A is a table showing Gene Set Enrichment Analysis (GSEA) of bulk RNA-sequencing data from Day 15 skin biopsies in the NHP imiquimod (IMQ) model comparing afucosylated Ab-1 (10 mg / kg)-treated animals to vehicle controls. FIG. 22B is a series of graphs showing Gene Set Variation Analysis (GSVA) quantification of pathway- specific enrichment scores for each individual sample (afucosylated Ab-1 treatment or vehicle control). “Ab-1” in FIG.22B refers to afucosylated Ab-1.

[0135] FIG. 23 is a schematic showing the design of the IMQ study.

[0136] FIG. 24 is a FACS plot showing the number of CD 161+ CD3+ T cells from skin biopsies taken at day 21 from the indicated treatment group (vehicle control; 3 x dose of lOmg / kg afucosylated Ab-1 treatment group; and single dose of 30mg / kg afucosylated Ab-1 treatment group).

[0137] FIG. 25 is a bar graph showing the absolute counts of CD161+ T cells in the skin at day 21 from the indicated treatment group (vehicle control; 3 x dose of lOmg / kg afucosylated Ab-1 treatment group; and single dose of 30mg / kg afucosylated Ab-1 treatment group).

[0138] FIG. 26 is a bar graph showing the percent reduction in CD 161+ T cells in the skin at day 21 from the indicated treatment group (vehicle control; 3 x dose of lOmg / kg afucosylated Ab-1 treatment group; and single dose of 30mg / kg afucosylated Ab-1 treatment group).

[0139] FIG.27 is a series of line graphs showing the PASI score over 1-21 days in each of the indicated treatment groups (vehicle control; 3 x dose of lOmg / kg treatment group; and single doseAttorney Docket No. 63340.32WO01of 30mg / kg treatment group). “Ab-1” in FIG. 27 refers to afucosylated Ab-1.

[0140] FIG. 28 is a heat map showing the change in expression level of the indicated gene (cytokine, chemokine. or psoriasis skin lesion associated gene) in skin biopsies from cynomolgus monkeys in the IMQ mediated Psoriasis model from the indicated treatment group (vehicle control; 3 x dose of lOmg / kg treatment group; and single dose of 30mg / kg treatment group) at day 21.

[0141] FIG. 29A is a table showing Gene Set Enrichment Analysis (GSEA) of bulk RNA-sequencing data from Day 21 skin biopsies in the NHP imiquimod (IMQ) model comparing Ab-1-treated animals to vehicle controls. FIG. 29B is a series of graphs showing Gene Set Variation Analysis (GSVA) quantification of pathway- specific enrichment scores for each individual sample (Afucosylated Ab-1 treatment or vehicle control). “Ab-1” in FIG. 29B refers to afucosylated Ab-1.

[0142] FIG. 30A is a dot graph showing the number of neutrophils in skin sample at day 21 of animals treated with vehicle control or afucosylated Ab-1; along with bar graphs showing the level of CXCL1 and CXCL8 at day 7 and day 21 in the indicated treatment group (vehicle control or afucosylated Ab-1). FIG.30B is a dot graph showing the number of macrophages in skin sample at day 21 of animals treated with vehicle control or afucosylated Ab-1; along with bar graph showing the level of CCL2 / MCP1 and CXCL12 at day 7 and day 21 in the indicated treatment group (vehicle control or afucosylated Ab-1).

[0143] FIG. 31 is a heat map showing Gene Set Enrichment Analysis (GSEA) of bulk RNA-sequencing data from Day 21 skin biopsies in the NHP IMQ model comparing vehicle control, afucosylated Ab-l-treatment group, anti-IL-23 antibody treatment, and anti-TNF antibody treatment group. “Ab-1” in FIG. 31 refers to afucosylated Ab-1.

[0144] FIG. 32 is a series of graphs showing Gene Set Variation Analysis (GSVA) quantification of pathway-specific enrichment scores for each individual sample (vehicle control, afucosylated Ab-l-treatment group, anti-IL-23 antibody treatment, and anti-TNF antibody treatment group).

[0145] FIG. 33A is a plot and a bar graph showing the percentage of CD4+ CD 161+ T cells and CD8+ CD161+ T cells in samples of intestinal tissue from Crohn’s disease patients. FIG.33B is a series of bar graphs showing the level of IL-17F, IL-22, IFNy, and TNFa expressed by CD161-cells and CD 161+ cells in samples of intestinal tissue from Crohn’s disease patients.

[0146] FIG. 34 is a series of line graphs showing the concentration (pg / mL) of each of theAttorney Docket No. 63340.32WO01indicated cytokines or chemokines (Th 17 cytokines - TL17f, IL-22; Pro-inflammatory cytokines -GMCSF, IFNy, TNFa; and IL17 regulated chemokines - CXCL5, CCL20, and IL8) from samples of intestinal tissue from Crohn’s disease patients in the presence or absence of CD161+ cell depletion and in the indicated treatment group (no treatment, anti-CD3 / CD28 antibody treatment, anti-IL23 / IL-ip antibody treatment, or anti-IL-18 / IL-12 antibody treatment).

[0147] FIG. 35A is a line graph showing the binding affinity (MFI) of Ab-1. HP-3G10. and hlgGl control to human CD 161 expressed by HEK293 cells in vitro. FIG. 35B is a line graph showing the binding affinity (MFI) of Ab-1, HP-3G10, and hlgGl control to cyno CD 161 expressed by HEK.293 cells in vitro.

[0148] FIG. 36 is a graph showing the Octet epitope binning assay between Ab-1 and HP-3010 in sandwich format. A positive signal or shift in wavelength was observed from binding of HP3G10 to human CD 161 bound by immobilized Ab-1, which indicated that HP3G10 is not competing with Ab-1 for the same binding site, and therefore belong to different epitope bins.

[0149] FIG. 37 shows two FACS plots showing isotype control versus HP-3G10 binding to human CD 161 (left) and Ab-1 versus HP-3G10 binding to human CD 161 (right).

[0150] FIG. 38 is a series of FACS plots showing the phagocytosis of CD161- T cells and CD 161+ T cells in the indicated treatment group.

[0151] FIG. 39 is a bar graph showing the % phagocytosis of CD161- T cells and CD 161+ T cells in the indicated treatment group.

[0152] FIG. 40A is a bar graph showing the concentration of GMCSF (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor.FIG. 40B is a bar graph showing the concentration of IFNy (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG. 40C is a bar graph showing the concentration of IL-2 (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG.40D is a bar graph showing the concentration of IL-12p70 (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG.40E is a bar graph showingAttorney Docket No. 63340.32WO01the concentration of IL-6 (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG. 40F is a bar graph showing the concentration of IL-ip (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG. 40G is a bar graph showing the concentration of TNFa (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG. 40H is a bar graph showing the concentration of IL-17A (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor.FIG. 401 is a bar graph showing the concentration of IL-4 (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. FIG.40J is a bar graph showing the concentration of IL- 10 (pg / mL) produced by human whole blood cells treated with the indicated agent in vitro. All treatments were performed in triplicates per donor. Every data point is expressed as the average of triplicates for each donor. “Ab-1” in FIGS. 40A-40 J refers to afucosylated Ab-1.

[0153] FIG.41A is a bar graph showing the log2 fold change in IL-17F from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / F antibody, or anti-IgGl control) antibody. FIG.41B is a bar graph showing the log2 fold change in IL-6 from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / F antibody, or anti-IgGl control) antibody. FIG. 41C is a bar graph showing the log2 fold change in IL-22 from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / F antibody, or anti-IgGl control) antibody. FIG. 41D is a bar graph showing the log2 fold change in IFNy from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / F antibody, or anti-IgGl control) antibody. FIG.41E is a bar graph showing the log2 fold change in TNFa from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / FAttorney Docket No. 63340.32WO01antibody, or anti-TgGl control) antibody. FIG.41F is a bar graph showing the log2 fold change in GM-CSF from Crohn’s disease patient-derived colonic biopsies treated with the indicated agent (Ab-1, anti-IL-17A antibody, anti-TNF-a antibody, anti-IL-17A / F antibody, or anti-IgGl control) antibody.5. DETAILED DESCRIPTION

[0154] The inventors have, inter alia, engineered novel proteins that specifically bind CD 161. Accordingly, the novel CD 161 binding proteins disclosed herein are good candidates for the treatment of diseases {e.g., pro-inflammatory diseases (e.g., autoimmune diseases), cancer), diagnostics, etc. The inventors have further discovered, inter alia, that CD161 expressing immune cells e.g., activated immune cells e.g., T cells)) can be pathogenic, e.g., expressing higher levels of proinflammatory cytokines {e.g., within the context of a proinflammatory {e.g., autoimmune) disease). Without wishing to be bound by theory, in some embodiments, the CD16 binding proteins {e.g., anti-CD161 antibodies) described herein function to, inter alia, deplete CD161 expressing immune cells (e.g., pathogenic immune cells {e.g., activated immune cells {e.g., T cells))), thereby, e.g., reducing the level of one or more proinflammatory cytokine {e.g., reducing the level of a plurality of proinflammatory cytokines simultaneously). As such, the CD16 binding proteins (e.g., anti-CD161 antibodies) described herein are useful, inter alia, for the treatment of proinflammatory diseases {e.g., autoimmune diseases). As such, the current disclosure provides CD16 binding proteins {e.g., anti-CD161 antibodies) and their use in, inter alia, pharmaceutical compositions, and methods of treating diseases {e.g., proinflammatory diseases {e.g., autoimmune diseases)).TABLE OF CONTENTS5.1 Definitions5.2 CD161 Binding Proteins5.3 Ig Constant Regions5.3.1 Ig Effector Function5.3.1.1 Enhanced Ig Effector Function5.3.1.2 Reduced Ig Effector F unction5.3.2 Promotion of Heterodimerization5.4 Conjugates and Fusion ProteinsAttorney Docket No. 63340.32WO015.4.1 Chimeric Antigen Receptors5.4.2 T-Cell Engagers5.4.3 Half-Life Extension Moieties5.4.4 Linkers5.4.4.1 Non-peptide Linkers5.4.4.2 Peptide Linkers5.4.5 Orientation5.5 Multimeric CD161 Binding Proteins5.5.1 Exemplary Properties of CD161 Binding Proteins5.5.1.1 Affinity of CD161 Binding Proteins for CD1615.5.1.2 Inhibition of CD161 Binding to CLEC2D5.5.1.3 Depletion of Populations of CD161 Expressing Cells and Cytokines5.6 Methods of Making Proteins5.6.1 Methods of Making Afucosylated Antibodies5.7 Polynucleotides, Vectors, Carriers, & Cells5.8 Pharmaceutical Compositions5.9 Methods of Use5.9.1 Methods of Delivery5.9.2 Methods of Inhibiting Binding of CD161 to CLEC2D5.9.3 Methods of Treating a Disease5.9.4 Methods of Treating a Proinflammatory Disease5.9.5 Methods of Treating Autoimmune Diseases5.9.6 Methods of Depleting CD161 Expressing Cells5.9.7 Methods of Reducing the Level of Proinflammatory Cytokines5.9.8 Methods of Treating Cancer5.10 Kits5.1 Definitions

[0155] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0156] Unless defined otherwise, all technical and scientific terms used herein have the sameAttorney Docket No. 63340.32WO01meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed.

[0157] In this application, the use of the singular includes the plural unless specifically stated otherwise. For example, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0158] It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of’ and “consisting essentially of’ are also provided.

[0159] The term “and / or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” as used in a phrase such as “A and / or B” herein is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0160] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.

[0161] The terms “about” or “comprising essentially of’ refer to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. When particular values or compositions are provided in the application and claims, unless otherwise stated, the meaning of “about” or “comprising essentially of” should be assumed to be within an acceptable error range for that particular value or composition.

[0162] Where proteins are described herein, it is understood that polynucleotides (e.g., RNA or DNA nucleic acid molecules) encoding the proteins are also provided herein.Attorney Docket No. 63340.32WO01

[0163] Where proteins, nucleic acid molecules, vectors, carriers, etc. are described herein, it is understood that isolated forms of the proteins, nucleic acid molecules, vectors, carriers, etc. are also provided herein.

[0164] Where proteins, nucleic acid molecules, etc. are described herein, it is understood that recombinant forms of the proteins, nucleic acid molecules, etc. are also provided herein.

[0165] Where proteins or sets of proteins are described herein, it is understood that both proteins comprising the primary structure are provided herein as well as proteins folded into their three-dimensional structure (z.e., tertiary or quaternary structure) are provided herein.

[0166] As used herein, the term “antibody dependent cell mediated cytotoxicity” or “ADCC” refers to an immune mechanism leading to the lysis of antibody (or an Fc region containing protein) (e.g., an Ig Fc containing fusion protein described herein)-coated target cells by immune effector cells (e.g., NK cells). As used herein, the term “reduced ADCC” and the like refers to either a reduction in the number of target cells that are lysed in a given time, at a given concentration of antibody (or an Ig Fc region containing protein) (e.g., an Fc region containing fusion protein described herein) in the medium surrounding the target cells, by the mechanism of ADCC defined above, and / or an increase in the concentration of antibody (or an Fc region containing protein) (e.g., an Fc containing fusion protein described herein) in the medium surrounding the target cells, required to achieve the lysis of a given number of target cells in a given time, by the mechanism of ADCC defined above. The reduction in ADCC is relative to the ADCC mediated by the same antibody (or an Fc region containing protein) (e.g., an Fc containing fusion protein described herein) produced by the same type of host cells, using the same standard production, purification, formulation and storage methods (which are known to those skilled in the art), but that has not been engineered (e.g., does not comprise one or more amino acid variation, e.g., amino acid substitution, that mediates a decrease in ADCC). For example the reduction in ADCC mediated by an antibody (or an Fc region containing protein) (e.g., an Fc containing fusion protein described herein) comprising in its Fc region an amino acid substitution that reduces ADCC, is relative to the ADCC mediated by the same antibody (or an Fc region containing protein) (e.g., an Fc containing fusion protein described herein) without said amino acid substitution in the Fc region.

[0167] As used herein, the term “administering” refers to the physical introduction of an agent, e.g., a therapeutic agent (or a precursor of the therapeutic agent that is metabolized or altered within the body of the subject to produce the therapeutic agent in vivo) (e.g., a CD161 binding proteinAttorney Docket No. 63340.32WO01described herein (e.g., an anti-CD161 antibody described herein)) to a subject, using any of the various methods and delivery systems known to those skilled in the art. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods. The term administering includes self-administration and non-self-administration.

[0168] As used herein, the term “affinity” refers to the strength of the binding of one protein (e.g., an Antibody) to another protein (e.g., an Antigen). The affinity of a protein is measured by the dissociation constant Kd, defined as [Antibody] x [Antigen] / [Antibody- Antigen] where [Antibody-Antigen] is the molar concentration of the Antibody-Antigen complex, [Antibody] is the molar concentration of the unbound Antibody and [Antigen] is the molar concentration of the unbound Antigen. The affinity constant Ka is defined by 1 / Kd. Standard methods of measuring affinity are known to the person of ordinary skill in the art. Exemplary methods of measuring affinity are described herein, see for example, § 5.5.1.

[0169] As used herein, the terms “agent” is used generically to describe any macro or micro molecule. Exemplary moieties include, but are not limited polypeptides, proteins, peptides, polynucleotides (e.g., DNA, RNA), small molecules, carbohydrates, lipids, synthetic polymers (e.g., polymers of PEG) (or any combinations thereof).

[0170] As used herein, the term “antibody” or “antibodies” is used in the broadest sense and encompasses various immunoglobulin (Ig) (e.g., human Ig (hlg)) structures, including, but not limited to monoclonal antibodies, polyclonal antibodies, multispecific (e.g., bispecific, trispecific) antibodies, and antibody fragments so long as they exhibit the desired antigen-binding activity (i.e., antigen binding fragments or variants). It is common in the art to refer to an anti-X antibody, wherein X is the antigen (e.g., CD161). The term antibody thus includes, for example, full-length antibodies; antigen-binding fragments of full-length antibodies; molecules comprising antibody CDRs, VH regions, and / or VL regions; and antibody-like scaffolds (e.g., fibronectins). Examples of antibodies include, without limitation, monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies, human antibodies, humanized antibodies, chimeric antibodies, camelized antibodies, intrabodies, affybodies, diabodies, tribodies, heteroconjugate antibodies, antibody-drug conjugates, single domain antibodies (e.g.,VHH, (VHH)2), single chain antibodies, single-chain Fvs (scFv; (scFv)2), Fab fragments (e.g., Fab, single chain Fab (scFab), F(ab’)2 fragments, disulfide-linked Fvs (sdFv), Fc fusions (e.g., Fab-Fc, scFv-Fc, VHH-Fc, (SCFV)2-FC, (VHH)2-FC), and antigen-binding fragments of any of the above, andAttorney Docket No. 63340.32WO01conjugates or fusion proteins comprising any of the above. Antibodies can be of Ig isotype (e.g., IgG, IgE, IgM, IgD, or IgA), any class (e.g., IgGi, IgG2, IgGs, IgG4, IgAi or IgA2), or any subclass (e.g., IgG2a or IgGzh) of Ig). In certain embodiments, antibodies described herein are IgG antibodies, or a class (e.g., human IgGi or IgG4) or subclass thereof. In some embodiments, the antibody is a human, humanized, or chimeric IgGi or IgG4 monoclonal antibody. In some embodiments, the term antibodies refers to a monoclonal or polyclonal antibody population. Antibodies described herein can be produced by any standard methos known in the art, e.g., recombinant production in host cells, see, e.g., § 5.6; or synthetic production.

[0171] As used herein, the term “CD161” refers to the type II transmembrane C-type lectin-like receptor expressed, e.g., by natural killer cells. CD161 is also commonly referred to in the art as Killer cell lectin-like receptor subfamily B member 1 (KLRB1). The amino acid sequence of a reference human CD161 (hCD161) protein is set forth in SEQ ID NO: 1 (UniProt Ref.: Q12918-1).

[0172] As used herein, the term “CD161 binding protein” and the like refers to a protein (e.g., an antibody (i.e., an anti-CD161 antibody)) that specifically binds CD 161.

[0173] As used herein, the term “CLEC2D” or “C-type lectin domain family 2 member D” refers to the C type lectin receptor that, inter alia, binds histones released upon necrotic cell death. The amino acid sequence of a reference human CLEC2D (hCLEC2D) isoform is set forth in SEQ ID NO: 2 (UniProt Ref.: Q9UHP7-1). Multiple isoforms of hCLEC2D are known produced by alternative splicing.

[0174] As used herein, the term “CDR” or “complementarity determining region” refers to the noncontiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. These particular regions have been described by Kabat et al., J. Biol. Chem.252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991), the entire contents of each of which is incorporated herein by reference for all purposes. Unless otherwise specified, the term “CDR” is a CDR as defined by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat el al., Sequences of protein of immunological interest. (1991). A person of ordinary skill in the art would be able to determine the CDRs as defined by another scheme, e.g., Chothia, IMGT, using ordinary methods known in the art. The three sequential CDRs of the VH region are typically termed “CDR-H1”, “CDR-H2”, and “CDR-H3” herein. The threeAttorney Docket No. 63340.32WO01sequential CDRs of the VL region are typically termed “CDR-L1”, “CDR-L2”, and “CDR-L3” herein, as is common in the art.

[0175] The terms “CHI” and “CHI region” are used interchangeably herein and refer to the first constant region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference IgGl CHI region is set forth in SEQ ID NO: 11: and the amino acid sequence of an exemplary reference IgG4 CHI region is set forth in SEQ ID NO: 24.

[0176] The terms “CH2” and “CH2 region” are used interchangeably herein and refer to the second constant region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference IgGl CH2 region is set forth in SEQ ID NO: 13; and the amino acid sequence of an exemplary reference IgG4 CH2 region is set forth in SEQ ID NO: 26.

[0177] The terms “CH3” and “CH3 region” are used interchangeably herein and refer to the third constant region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference IgGl CH3 region is set forth in SEQ ID NO: 14; and the amino acid sequence of an exemplary reference IgG4 CH3 region is set forth in SEQ ID NO: 27.

[0178] As used herein, the term “conjugation” refers to chemical conjugation of a protein with a moiety (e.g., small molecule, polypeptide, nucleic acid molecule, carbohydrate, lipid, synthetic polymer (e.g., polymers of polyethylene glycol (PEG)), etc.). The moiety can be directly connected to the protein or indirectly connected through a linker, e.g., as described herein. Chemical conjugation methods are well known in the art, as are commercially available conjugation reagents and kits, with detailed instructions for their use readily available from the commercial suppliers.

[0179] The terms “constant region” and “constant domain” are used interchangeably herein and refer to a carboxyl terminal portion of a light and / or heavy chain of a full-length antibody which is not directly involved in binding of an antibody to antigen, but which can exhibit various effector functions, such as interaction with an Ig Fc receptor (e.g., Fc gamma receptor). The constant region of an Ig molecule generally has a more conserved amino acid sequence relative to an Ig variable domain.

[0180] As used herein, the term “combination” in reference to the administration of at least two agents to a subject or similar terms such as “administering in combination”, “coadministration” or “combination” includes the administration of at least two active agents. The two agents may be formulated in separate pharmaceutical formulations or in single pharmaceutical formulation. The at least two active agents may be administered simultaneously or consecutivelyAttorney Docket No. 63340.32WO01in any order such that, there is a time period while both (or all) active agents overlap in exerting their biological activities. Administering in combination does not require that the agents are administered at the same time, at the same frequency, or by the same route of administration.

[0181] As used herein, the term “derived from,” with reference to a polynucleotide refers to a polynucleotide that has at least 70% sequence identity to a reference polynucleotide {e.g., a naturally occurring polynucleotide) or a fragment thereof. The term “derived from,” with reference to a protein refers to a protein that comprises an amino acid sequence that has at least 70% sequence identity to the amino acid sequence of a reference protein {e.g., a naturally occurring protein). The term “derived from” as used herein does not denote any specific process or method for obtaining the polynucleotide or protein. For example, the polynucleotide or protein can be recombinantly produced or chemically synthesized.

[0182] As used herein, the term “disease” refers to any abnormal condition that impairs physiological function. The term is used broadly to encompass any disorder, illness, abnormality, pathology, sickness, condition, or syndrome in which physiological function is impaired, irrespective of the nature of the etiology.

[0183] The terms “DNA” and “polydeoxyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple deoxyribonucleotides that are polymerized via phosphodiester bonds. Deoxyribonucleotides are nucleotides in which the sugar is deoxyribose.

[0184] The term “effector function” when used in reference to an Ig Fc region or a protein comprising an Ig Fc region {e.g., a full-length antibody) refers to those biological activities attributable to the Ig Fc region of a typical full-length antibody, which therefore vary with the antibody isotype. Antibody effector functions include, but are not limited to. antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement dependent cytotoxicity (CDC). Fc receptor binding e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa. and / or Fey R II I b {e.g., FcyRI, Fcylla, and / or Fcyllla)). and Clq binding.

[0185] As used herein, the term “EU numbering system” refers to the EU numbering convention for the constant regions of an antibody, as described in Edelman, G.M. el al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat et al, Sequences of Proteins of Immunological Interest, U.S. Dept. Health and Human Services, 5th edition, 1991, the entire contents of each of which is incorporated herein by reference for all purposes.Attorney Docket No. 63340.32WO01

[0186] As used herein, the term “Fab” refers to an antigen binding domain that comprises a Fab heavy chain that comprises from N- to C-terminus a VH region and a CHI region; and a light chain comprising from N- to C-terminus a VL region and a CL region; and wherein the Fab heavy chain and the light chain associate to form an antigen binding domain.

[0187] The term “Fab-Fc” as used herein refers to an antibody that comprises a Fab operably linked to an Fc region.

[0188] As used herein, the term “Fc region” refers to the C-terminal region of an Ig heavy chain that comprises from N- to C-terminus at least a CH2 region operably connected to a CH3 region. In some embodiments, the Fc region comprises an Ig hinge region or at least a portion of an Ig hinge region operably connected to the N-terminus of the CH2 region. In some embodiments, the Fc region is engineered relative to a reference Fc region. Additional examples of proteins with engineered Fc regions can be found in Saunders 2019 (K. O. Saunders, “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life,” 2019, Frontiers in Immunology, V. 10. Art. 1296, pp. 1-20. the entire contents of which is incorporated by reference herein for all purposes).

[0189] As used herein, the terms “first” and “second” with respect to Fc regions etc., are used for convenience of distinguishing when there is more than one of each type of moiety. Use of these terms is not intended to confer a specific order or orientation in the protein unless explicitly so stated. For example, an antibody described herein (e.g., in the case of a full-length antibody) may contain two Fc regions that associate e.g., via one or more covalent (e.g., disulfide) bond.

[0190] As used herein, the term “framework region” or “FR region” refers to the amino acid residues that are part of the variable region of an antibody, but are not part of the CDRs e.g., using the Kabat definition of CDRs).

[0191] As used herein, the term “full-length antibody” refers to an antibody having a structure substantially similar to a native antibody structure (i) a first immunoglobulin (Ig) light chain comprising from N- to C-terminus a light chain variable region (VL) region and a light chain constant region (CL) region; (ii) a first Ig heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N- to C-terminus a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chainAttorney Docket No. 63340.32WO01associate to form a first antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the two heavy chains comprise a substantially identical amino acid sequence; and the two light chains comprise a substantially identical amino acid sequence. In some embodiments, the two heavy chains comprise a substantially identical amino acid sequence except for one or more amino acid modifications that promote heterodimerization of the correct heavy chains (e.g., as described herein); and the two light chains comprise a substantially identical amino acid sequence. Antibody chains may be substantially identical but not entirely identical if they differ due to post-translational modifications, such as C-terminal cleavage of lysine residues, alternative glycosylation patterns, etc.

[0192] The term “functional variant” as used herein in reference to a protein refers to a protein that comprises at least one but no more than 15%, not more than 12%, no more than 10%, no more than 8% amino acid modification (e.g., substitution, deletion, addition) compared to the amino acid sequence of a reference protein, wherein the protein retains at least one particular function of the reference protein. Not all functions of the reference protein (e.g., wild type) need be retained by the functional variant of the protein. In some instances, one or more functions are selectively reduced or eliminated. In some embodiments, the reference protein is a wild type protein. For example, a functional variant of an antibody that specifically binds CD 161 can refer to the antibody that specifically binds CD161 comprising one or more amino acid substitution as compared to a reference antibody that retains the ability to specifically bind CD 161.

[0193] The term “functional fragment” as used herein in reference to a protein refers to a fragment of a reference protein that retains at least one particular function. Not all functions of the reference protein need be retained by a functional fragment of the protein. In some instances, one or more functions are selectively reduced or eliminated. In some embodiments, the reference protein is a wild type protein. In some embodiments, the functional fragment protein comprises at least one but no more than 15%, not more than 12%, no more than 10%, no more than 8% amino acid deletion compared to the amino acid sequence of a reference protein. For example, a functional fragment of an antibody that specifically binds CD 161 can refer to a fragment of the antibody that retains the ability to specifically bind CD 161.

[0194] As used herein, the term “fuse” and grammatical equivalents thereof refers to theAttorney Docket No. 63340.32WO01operable connection of at least a first polypeptide to a second polypeptide, wherein the first and second polypeptides are not naturally found operably connected together. For example, the first and second polypeptides are derived from different proteins and / or are from different organisms. The term fuse encompasses both a direct connection of the at least two polypeptides through a peptide bond, and the indirect connection through a linker (e.g., a peptide linker).

[0195] As used herein, the term “fusion protein” and grammatical equivalents thereof refer to a protein that comprises at least one polypeptide operably connected to another polypeptide, wherein the first and second polypeptides are not naturally found operably connected together. For example, the first and second polypeptides of the fusion protein are each derived from different proteins and / or are from heterologous organisms. For the sake of clarity, it will be understood that neither the first nor second polypeptide is required to be a full-length protein (e.g., a full-length naturally occurring protein). For example, the first and / or second polypeptide can comprise or consist of fragments (e.g., functional fragments or domains of full-length proteins (e.g., engineered, naturally occurring). The at least two polypeptides of the fusion protein can be directly operably connected through a peptide bond; or can be indirectly operably connected through a linker (e.g., a peptide linker). Thus, the term fusion polypeptide encompasses embodiments, wherein Polypeptide A is directly operably connected to Polypeptide B through a peptide bond (Polypeptide A - Polypeptide B), and embodiments, wherein Polypeptide A is operably connected to Polypeptide B through a peptide linker (Polypeptide A - peptide linker - Polypeptide B).

[0196] As used herein, the term “half-life extension moiety” refers to a moiety (e.g., small molecule, polypeptide, nucleic acid molecule, carbohydrate, lipid, synthetic polymer (e.g., polymers of PEG), etc.) that when conjugated or otherwise operably connected (e.g., fused) to a protein (the subject protein), increases the half-life of the subject protein in vivo when administered to a subject (e.g., a human subject). The pharmacokinetic properties of the protein can be evaluated utilizing in vivo models known in the art.

[0197] As used herein, the term “half-life extension polypeptide” or “half-life extension protein” refers to a protein that when operably connected to another protein (the subject protein), increases the half-life of the subject protein in vivo when administered to a subject (e.g., a human subject). The pharmacokinetic properties of the protein can be evaluated utilizing in vivo models known in the art.Attorney Docket No. 63340.32WO01

[0198] As used herein, the term “heterologous”, when used to describe a first element in reference to a second element means that the first element and second element do not exist in nature disposed as described. For example, a polypeptide comprising a “heterologous moiety” means a polypeptide that is joined to a moiety (e.g., small molecule, polypeptide, nucleic acid molecule, carbohydrate, lipid, synthetic polymer (e.g., polymers of PEG), etc.) that is not joined to the polypeptide in nature.

[0199] As used herein, the term “heavy chain” refers to the portion of an immunoglobulin (e.g., a human Ig) that typically comprises from N- to C-terminus a heavy chain variable region (VH), a CHI region, a hinge region, a CH2 region, and a CH3 region. The constant regions of the heavy chain (i.e., the CHI region, the hinge region, the CH2 region, and the CH3 region) can be any distinct isotype, for example, human alpha (a), delta (8), epsilon (s), gamma (y), and mu (p), based on the amino acid sequence of the constant domain, which give rise to the IgA, IgD, IgE, IgG, and IgM classes of human antibodies, respectively, including subclasses of IgG, e.g., IgGi, IgG2, IgG?. and IgG4. As used herein, the term “heavy chain” when used in reference to a human antibody can refer to any distinct type, e.g., alpha (a), delta (8), epsilon (s), gamma (y), and mu (p), based on the amino acid sequence of the constant domain, which give rise to human IgA, IgD, IgE, IgG, and IgM classes of antibodies, respectively, including subclasses of human IgG, e.g., IgGi, IgG2, IgG?, and IgG4.

[0200] The terms “hinge” or “hinge region” are used interchangeably herein and refer to the hinge region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference IgGi hinge region is set forth in SEQ ID NO: 12; and the amino acid sequence of an exemplary reference IgG4 hinge region is set forth in SEQ ID NO: 25.

[0201] As used herein, the term “isolated” with reference to a protein or polynucleotide refers to a protein or polynucleotide that is substantially free of other cellular components with which it is associated in the natural state.

[0202] As used herein, the term “moiety” is used generically to describe any macro or micro molecule that can be incorporated into a fusion protein described herein. The moieties of a fusion protein are operably connected. Exemplary moieties include, but are not limited protein, polypeptides, polynucleotides (e.g., DNA, RNA), small molecules, carbohydrates, lipids, synthetic polymers (e.g., polymers of PEG). In some embodiments, the moiety is a protein.

[0203] As used herein, the term “operably connected” refers to the linkage of two moieties inAttorney Docket No. 63340.32WO01a functional relationship. For example, a polypeptide is operably connected to another polypeptide when they are linked (either directly or indirectly via a peptide linker) in frame such that both polypeptides are functional (e.g., a fusion protein described herein). Or for example, a transcription regulatory polynucleotide e.g., a promoter, enhancer, or other expression control element is operably linked to a polynucleotide that encodes a protein if it affects the transcription of the polynucleotide that encodes the protein. The term “operably connected” can also refer to the conjugation of a moiety to e.g., a polynucleotide or polypeptide (e.g., the conjugation of a PEG polymer to a protein).

[0204] The determination of “percent identity” between two sequences (e.g., peptide or protein (amino acid sequences) or polynucleotide (nucleic acid sequences)) can be accomplished using a mathematical algorithm. A specific, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, each of which is herein incorporated by reference in its entirety. Such an algorithm is incorporated into the NBEAST and XBEAST programs of Altschul SF et al., (1990) J Mol Biol 215: 403, which is herein incorporated by reference in its entirety. BEAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecule described herein. BEAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is herein incorporated by reference in its entirety. Alternatively, PSI BEAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is herein incorporated by reference in its entirety. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acidAttorney Docket No. 63340.32WO01sequences, a PAM 120 weight residue table, a gap length penalty of 1 , and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.

[0205] As used herein, the term “pharmaceutical composition” means a composition that is suitable for administration to an animal, e.g., a human subject, and comprises a therapeutic agent and a pharmaceutically acceptable carrier or diluent. A “pharmaceutically acceptable carrier or diluent” means a substance intended for use in contact with the tissues of human beings and / or non-human animals, and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable therapeutic benefit / risk ratio.

[0206] As used herein, the term “plurality” means 2 or more (e.g., 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 9 or more, or 10 or more).

[0207] The terms “polynucleotide” and “nucleic acid molecule” are used interchangeably herein and refer to a polymer of DNA or RNA. The nucleic acid molecule can be single- stranded or double- stranded; contain natural, non-natural, or altered nucleotides; and contain a natural, nonnatural, or altered internucleotide linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule. Nucleic acid molecules include, but are not limited to, all nucleic acid molecules which are obtained by any means available in the art, including, without limitation, recombinant means, e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and by synthetic means. The skilled artisan will appreciate that, except where otherwise noted, nucleic acid sequences set forth in the instant application will recite thymidine (T) in a representative DNA sequence but where the sequence represents RNA (e.g., mRNA), the thymidines (Ts) would be substituted for uracils (Us). Thus, any of the RNA polynucleotides encoded by a DNA identified by a particular sequence identification number may also comprise the corresponding RNA (e.g., mRNA) sequence encoded by the DNA, where each thymidine (T) of the DNA sequence is substituted with uracil (U).

[0208] As used herein, the terms “protein” and “polypeptide” refer to a polymer of at least 2 (e.g., at least 5) amino acids linked by a peptide bond. The term “polypeptide” does not denote a specific length of the polymer chain of amino acids. It is common in the art to refer to shorterAttorney Docket No. 63340.32WO01polymers of amino acids (e.g., approximately 2-50 amino acids) as peptides; and to refer to longer polymers of amino acids (e.g., approximately over 50 amino acids) as polypeptides. However, the terms “peptide” and “polypeptide” and “protein” are used interchangeably herein. In some embodiments, a protein is folded into its three-dimensional structure. Where proteins are contemplated herein, it should be understood that proteins comprising the primary structure are provided herein as well as proteins folded into their three-dimensional structure (i.e., tertiary or quaternary structure) are provided herein.

[0209] A “prophylactic” treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing pathology.

[0210] The terms “RNA” and “polyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple ribonucleotides that are polymerized via phosphodiester bonds. Ribonucleotides are nucleotides in which the sugar is ribose. RNA may contain modified nucleotides; and contain natural, non-natural. or altered internucleotide linkages, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule.

[0211] The term “scFv” or “single chain variable fragment” as used herein refers an antigen binding protein that comprises a VH operably connected (e.g., via a peptide linker) to a VL. In some embodiments, the VH is operably connected to the VL via a peptide linker. The components of the scFv can be in any orientation, for example, the scFv can comprise from N- to C-terminus a VH, a peptide linker, and a VL; or from N- to C-terminus a VL, a peptide linker, and a VH.

[0212] The term “(scFv ’ as used herein refers to an antibody that comprises a first and a second scFv operably connected (e.g., via a peptide linker). The first and second scFv can specifically bind the same or different antigens. In some embodiments, the first and second scFv are operably connected by a peptide linker.

[0213] The term “scFv-Fc” as used herein refers to an antibody that comprises a scFv operably linked (e.g., via a peptide linker) to an Fc domain or subunit of an Fc domain. In some embodiments, a scFv is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first scFv is operably connected to a first Fc domain and a second scFv is operably connected to a second Fc domain of a first and second Fc domain pair.

[0214] The term “(scFv)2-Fc” as used herein refers to a (scFv)2 operably linked (e.g., via aAttorney Docket No. 63340.32WO01peptide linker) to an Fc domain or a subunit of an Fc domain. Tn some embodiments, a (scFv)2 is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first (scFv)2 is operably connected to a first Fc domain and a second (scFv)2 is operably connected to a second Fc domain of a first and second Fc domain pair.

[0215] As used herein, the term “single domain antibody” or “sdAb” refers to an antibody having a single monomeric variable antibody domain. A sdAb is able to specifically bind to a specific antigen. A VHH (as defined herein) is an example of a sdAb.

[0216] As used herein, the term “specifically binds” with reference to two proteins refers to a preferential interaction, i.e., significantly higher binding affinity, between a first protein {e.g., an antibody) and a second protein {e.g., an antigen) relative to other amino acid sequences. Herein, when a first protein is said to “specifically bind” to a second protein, it is understood that the first protein specifically binds to an epitope of the second protein. The term “epitope” refers to the portion of the second protein that the first protein specifically recognizes. The term specifically binds includes molecules that are cross reactive with the same epitope of a different species. For example, an antibody that specifically binds human CD161 may be cross reactive with CD161 of another species {e.g., cynomolgus, murine, etc.) and still be considered herein to specifically bind human CD 161. The same term can be utilized to describe the preferential binding of non-protein agents {e.g., binding of a small molecule to a protein).

[0217] As used herein, the term “subject” includes any animal, such as a human or other animal. In some embodiments, the subject is a vertebrate animal e.g., mammal, bird, fish, reptile, or amphibian). In some embodiments, the subject is a human. In some embodiments, the method subject is a non-human mammal. In some embodiments, the subject is a non-human mammal is such as a non-human primate {e.g., monkeys, apes), ungulate {e.g., cattle, buffalo, sheep, goat, pig, camel, llama, alpaca, deer, horses, donkeys), carnivore {e.g., dog, cat), rodent {e.g., rat. mouse), or lagomorph {e.g., rabbit). In some embodiments, the subject is a bird, such as a member of the avian taxa Galliformes {e.g., chickens, turkeys, pheasants, quail), Anseriformes {e.g., ducks, geese), Paleaognathae {e.g., ostriches, emus), Columbiformes {e.g., pigeons, doves), or Psittaciformes {e.g., parrots).

[0218] As used herein, the term “therapeutically effective amount” of an agent {e.g., therapeutic agent) refers to any amount of the agent {e.g., therapeutic agent) that, when used alone or in combination with another therapeutic agent, improves a disease condition, e.g., protects aAttorney Docket No. 63340.32WO01subject against the onset of a disease (or infection); improves a symptom of disease or infection, e.g., decreases severity of disease or infection symptoms, decreases frequency or duration of disease or infection symptoms, increases disease or infection symptom-free periods; prevents or reduces impairment or disability due to the disease or infection; or promotes disease (or infection) regression. The ability of an agent (e.g., therapeutic agent) to improve a disease condition can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.

[0219] As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disease or infection and / or symptom(s) associated therewith or obtaining a desired pharmacologic and / or physiologic effect. It will be appreciated that, although not precluded, treating a disease or infection does not require that the disease, or symptom(s) associated therewith be completely eliminated. In some embodiments, the effect is therapeutic, i.e., without limitation, the effect partially or completely reduces, diminishes, abrogates, abates, alleviates, decreases the intensity of, or cures a disease and / or adverse symptom attributable to the disease. In some embodiments, the effect is preventative, i.e., the effect protects or prevents an occurrence or reoccurrence of a disease. To this end, the presently disclosed methods comprise administering a therapeutically effective amount of a composition as described herein. The term treating includes prophylactic use.

[0220] As used herein, the term “modification,” with reference to a polynucleotide, refers to a polynucleotide that comprises at least one substitution, alteration, inversion, addition, or deletion of nucleotide compared to a reference polynucleotide (e.g.,. one or more amino acid substitutions). Modifications can include the inclusion of non-naturally occurring nucleotide residues. As used herein, the term “modification,” with reference to an amino acid sequence refers to an amino acid sequence that comprises at least one substitution, alteration, inversion, addition, or deletion of an amino acid residue compared to a reference amino acid sequence. Modifications can include the inclusion of non-naturally occurring amino acid residues. Naturally occurring amino acid derivatives are not considered modified amino acids for purposes of determining percent identity of two amino acid sequences. For example, a naturally occurring modification of a glutamate amino acid residue to a pyroglutamate amino acid residue would not be considered an amino acid modification for purposes of determining percent identity of two amino acid sequences. Further,Attorney Docket No. 63340.32WO01for example, a naturally occurring modification of a glutamate amino acid residue to a pyroglutamate amino acid residue would not be considered an amino acid “modification” as defined herein.

[0221] As used herein, the term “variable region” refers to a portion of an antibody, generally, a portion of a light or heavy chain, typically about the amino-terminal 110 to 120 amino acids or 110 to 125 amino acids in the mature heavy chain and about 90 to 115 amino acids in the mature light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of a particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called complementarity determining regions (CDRs) while the more highly conserved regions in the variable domain are called framework regions (FR). Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In particular embodiments, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0222] The terms “VL” and “VL domain” are used interchangeably to refer to the light chain variable region of an antibody.

[0223] The terms “VH” and “VH domain” are used interchangeably to refer to the heavy chain variable region of an antibody.

[0224] The term “VHH” as used herein refers to a type of single domain antibody (sdAb) that has a single monomeric heavy chain variable antibody domain (VH). Such antibodies can be found in or produced from camelid mammals (e.g., camels, llamas) which are naturally devoid of light chains or synthetically produced.

[0225] The term “(VHH)2” as used herein refers to an antibody that comprises a first and a second VHH operably connected (e.g., via a peptide linker). The first and the second VHH can specifically bind the same or different antigens. In some embodiments, the first and second VHH are operably connected by a peptide linker.

[0226] The term “VHH-Fc” as used herein refers to an antibody that comprises a VHH operably linked (e.g., via a peptide linker) to an Fc domain or a subunit of an Fc domain. In someAttorney Docket No. 63340.32WO01embodiments, a VHH is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first VHH is operably connected to a first Fc domain and a second VHH is operably connected to a second Fc domain of a first Fc and a second Fc pair.

[0227] The term “(VHH)2-Fc” as used herein refers to (VHH) operably linked (e.g., via a peptide linker) to an Fc domain or a subunit of an Fc domain. In some embodiments, a (VHH)2 is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first (VHH)2 is operably connected to a first Fc domain and a second (VHH)2 is operably connected to a second Fc domain of a first Fc and a second Fc pair.5.2 CD161 Binding Proteins

[0228] As described above, provided herein are, inter alia, proteins e.g., antibodies (and functional fragments and functional variants thereof) that specifically bind CD161 (e.g., hCD161).

[0229] CD161 is a C-type lectin-like receptor, which is expressed e.g., on NK cells and subsets of both CD4+ and CD8+ T cells. CD161 binds, e.g., CLEC2D. CLEC2D is expressed, e.g., on the surface of both malignant cells and immune cells including germinal center B cells, activated T cells and tumor associated macrophages. CD161 is also expressed on pathogenic effector Th2 (peTh2 cells), TH2A cells, and ILC2 cells (innate like cell-2) that are involved in Th2 mediated diseases (including, e.g., allergies, asthma and eosinophilic diseases). See, e.g., Nakayama T, Hirahara K, Onodera A, Endo Y, Hosokawa H, Shinoda K, Tumes DJ, Okamoto Y. Th2 Cells in Health and Disease. Annu Rev Immunol. 2017 Apr 26;35:53-84. doi: 10.1146 / annurev-immunol-051116-052350. Epub 2016 Nov 28. PMID: 27912316; Wambre E, Bajzik V. DeLong JH. et al. A phenotypically and functionally distinct human TH2 cell subpopulation is associated with allergic disorders. Sci Transl Med. 2017;9(401):eaam9171. doi:10.1126 / scitranslmed.aam9171; and Morgan DM, Ruiter B, Smith NP, et al. Clonally expanded. GPR15-expressing pathogenic effector TH2 cells are associated with eosinophilic esophagitis. Sci Immunol. 2021;6(62):eabi5586. doi:10.1126 / sciimmunol.abi5586, the entire contents of each of which is incorporated herein by reference for all purposes.

[0230] The amino acid sequence of a reference hCD161 protein is set forth in SEQ ID NO: 1. Multiple isoforms of hCLEC2D are known produced by alternative splicing, with isoform 1 being the only isoform predominantly expressed at the cell surface. The amino acid sequence of a reference hCLEC2D isoform 1 protein is set forth in SEQ ID NO: 2. See Table 1, herein.Attorney Docket No. 63340.32WO01Table 1. The Amino Acid Sequence of a Reference hCLEC2D and hCD161 Protein.

[0231] In some embodiments, the CD161 binding protein comprises an antibody. In some embodiments, the CD 161 binding protein comprises a full-length antibody, Fab. Fab', F(ab')2, Fab-Fc, scFv, scFv-Fc, (scFv)2-Fc, Fv, a single domain antibody (sdAb) (e.g., a VHH), a sdAb-Fc (e.g., a VHH-Fc), (sdAb)2 (e.g., a (VHH)2, or a (sdAb)2-Fc (e.g., (VHH)2-Fc). In some embodiments, the CD161 binding protein comprises a full-length antibody, Fab, Fab', F(ab')2, Fab-Fc, scFv, scFv-Fc, (SCFV)2-FC, sdAb-Fc (e.g., a VHH-Fc), or (sdAb)2-Fc (e.g., (VHH)2-Fc). In some embodiments, the CD161 binding protein comprises a full-length antibody. In some embodiments, the CD 161 binding protein comprises a Fab. In some embodiments, the antibody comprises a F(ab')2. In some embodiments, the CD161 binding protein comprises a Fab-Fc. In some embodiments, the antibody comprises a scFv-Fc. In some embodiments, the CD 161 binding protein comprises a (scFv)2-Fc. In some embodiments, the CD161 binding protein comprises a sdAb-Fc (e.g., a VHH-Fc). In some embodiments, the CD161 binding protein comprises a (sdAb)2-Fc (e.g., (VHH>2-FC). In some embodiments, the CD161 binding protein comprises a single domain antibody. In some embodiments, the CD 161 binding protein comprises a VHH.

[0232] In some embodiments, the CD 161 binding protein comprises an antibody. In some embodiments, the antibody is an IgGI, IgG2, IgG3, or IgG4 antibody. In some embodiments, the antibody is an IgGI or IgG4 antibody. In some embodiments, the antibody is an IgGI antibody. In some embodiments, the antibody is an IgG4 antibody. In some embodiments, the antibody is an IgGI, IgG2, IgG3, or IgG4 antibody. In some embodiments, the antibody is an IgGI or IgG4Attorney Docket No. 63340.32WO01antibody. In some embodiments, the antibody is an TgGl antibody. Tn some embodiments, the antibody is an IgG4 antibody.

[0233] In specific preferred embodiments, the CD 161 binding protein is a full length antibody. In specific preferred embodiments, the CD 161 binding protein is a full length afucosylated antibody. In specific preferred embodiments, the CD 161 binding protein comprises a full-length antibody comprising (i) a first Ig light chain comprising from N- to C-terminus a light chain variable region (VL) region and a light chain constant region (CL) region; (ii) a first Ig heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N-to C-terminus a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chain associate to form a first antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the amino acid sequence of the first heavy chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the first light chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the VH region of the first heavy chain is 100% identical to the amino acid sequence of the VH region of the second heavy chain; and the amino acid sequence of the first heavy chain outside of the VH region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain outside of the VH region of the second heavy chain. In some embodiments, the amino acid sequence of the VL region of the first light chain is 100% identical to the amino acid sequence of the VL region of the second light chain; and the amino acid sequence of the first light chain outside of the VL region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second light chain outside of the VL region of the second light chain.

[0234] In specific preferred embodiments, the CD161 binding protein is an afucosylated full-length antibody comprising (i) a first Ig light chain comprising from N- to C-terminus a light chainAttorney Docket No. 63340.32WO01variable region (VL) region and a light chain constant region (CL) region; (ii) a first Tg heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N-to C-terminus a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chain associate to form a first antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the amino acid sequence of the first heavy chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the first light chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the VH region of the first heavy chain is 100% identical to the amino acid sequence of the VH region of the second heavy chain; and the amino acid sequence of the first heavy chain outside of the VH region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain outside of the VH region of the second heavy chain. In some embodiments, the amino acid sequence of the VL region of the first light chain is 100% identical to the amino acid sequence of the VL region of the second light chain; and the amino acid sequence of the first light chain outside of the VL region is at least 85%, 86%, 87%, 88%, 89%. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second light chain outside of the VL region of the second light chain.

[0235] In specific preferred embodiments, the CD161 binding protein is a monospecific afucosylated full-length antibody comprising (i) a first Ig light chain comprising from N- to C-terminus a light chain variable region (VL) region and a light chain constant region (CL) region; (ii) a first Ig heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N- to C-terminus a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chain associate to form a firstAttorney Docket No. 63340.32WO01antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the amino acid sequence of the first heavy chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the first light chain is at least 85%, 86%, 87%. 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the VH region of the first heavy chain is 100% identical to the amino acid sequence of the VH region of the second heavy chain; and the amino acid sequence of the first heavy chain outside of the VH region is at least 85%, 86%. 87%, 88%, 89%, 90%, 91%, 92%. 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain outside of the VH region of the second heavy chain. In some embodiments, the amino acid sequence of the VL region of the first light chain is 100% identical to the amino acid sequence of the VL region of the second light chain; and the amino acid sequence of the first light chain outside of the VL region is at least 85%, 86%, 87%. 88%, 89%, 90%, 91%. 92%, 93%, 94%, 95%. 96%, 97%, 98%, 99%. or 100% identical to the amino acid sequence of the second light chain outside of the VL region of the second light chain.

[0236] In some embodiments, the CD161 binding protein is monospecific. In some embodiments, the CD161 binding protein is multispecific (e.g., bispecific, trispecific). In some embodiments, the CD 161 binding protein is multispecific comprising at least one moiety that specifically binds another antigen (i.e., not CD161). In some embodiments, the CD161 binding protein is bispecific comprising at least one moiety that specifically binds another antigen (i.e., not CD161).

[0237] In some embodiments, the CD161 binding proteins (e.g., described herein) (or conjugates or fusions comprising the same) are multimeric (e.g., dimeric, trimeric, tetrameric, etc.) proteins comprising at least two, three, or four polypeptides.

[0238] In some embodiments, the CD 161 binding protein comprises at least two, three, or four polypeptides. In some embodiments, the CD161 binding protein is dimeric (i.e., comprises two polypeptides). In some embodiments, the CD 161 binding protein is trimeric (i.e., comprises threeAttorney Docket No. 63340.32WO01polypeptides). In some embodiments, the CD161 binding protein is tetrameric ( / ., comprises four polypeptides).

[0239] In some embodiments, two of the polypeptides associate via covalent or non-covalent interactions. In some embodiments, two of the polypeptides associate via at least one covalent interaction. In some embodiments, two of the polypeptides associate via one or more disulfide bond. In some embodiments, two of the polypeptides associate via 1, 2, 3, 4, or more disulfide bonds.

[0240] The amino acid sequence of hCD161 binding proteins (e.g., anti-hCD161 antibodies) of the present disclosure is provided in Table 2. The CDRs of the antibodies in Table 2, are denoted according to Kabat, Chothia, AbM, and IMGT. A person of ordinary skill in the art would be able to determine the CDRs as defined by another scheme, e.g., contact, using ordinary methods known in the art.Table 2. The Amino Acid Sequence of hCD161 Binding Proteins.Attorney Docket No. 63340.32WO01

[0241] In specific embodiments, the heavy chain lacks the C-terminal lysine at position 448, EU numbering according to Kabat. In some embodiments, the heavy lacks the C-terminal glycine and lysine at positions 447 and 448, respectively, EU numbering according to Kabat.

[0242] In some embodiments, the CD161 binding protein (e.g., anti-CD161 antibody) comprises a VH that comprises: a CDR-H1, a CDR-H2, and a CDR-H3. In some embodiments, the CD161 binding protein comprises a VL that comprises: CDR-L1, CDR-L2, and CDR-L3. In some embodiments, the CD161 binding protein comprises a VH that comprises: a CDR-H1, a CDR-H2, and a CDR-H3; and a VL that comprises: CDR-L1, CDR-L2, and CDR-L3.

[0243] In some embodiments, the CD161 binding protein comprises a CD161 binding protein provided in Table 2.

[0244] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acidAttorney Docket No. 63340.32WO01sequence of a VH CDR 1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0245] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2Attorney Docket No. 63340.32WO01comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0246] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH set forth in Table 2; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2; the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2.

[0247] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0248] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.);Attorney Docket No. 63340.32WO01the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.): the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0249] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).Attorney Docket No. 63340.32WO01

[0250] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH set forth in Table 2; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2; the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2.

[0251] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.): the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0252] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%. 86%, 87%, 88%, 89%, 90%. 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%. 92%, 93%, 94%. 95%, 96%, 97%. 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acidAttorney Docket No. 63340.32WO01sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%. 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises the amino acid sequence of a VL set forth in Table 2.

[0253] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%. 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of the amino acid sequence of a VL set forth in Table 2.

[0254] In some embodiments, the CD 161 binding protein comprises a light chain (LC) and a heavy chain (HC).

[0255] In some embodiments, the amino acid sequence of the HC comprises an amino acid sequence at least 85%. 86%, 87%, 88%, 89%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HCAttorney Docket No. 63340.32WO01comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC comprises an amino acid sequence at least 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HC comprises an amino acid sequence at least 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HC comprises the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC comprises the amino acid sequence of a LC set forth in Table 2.

[0256] In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC consists of an amino acid sequence at least 95%. 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence of a LC set forth in Table 2. In some embodiments, the amino acid sequence of the HC consists of the amino acid sequence of a HC set forth in Table 2; and the amino acid sequence of the LC consists of the amino acid sequence of a LC set forth in Table 2.

[0257] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the aminoAttorney Docket No. 63340.32WO01acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0258] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion,Attorney Docket No. 63340.32WO01addition, etc.).

[0259] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2; the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2.

[0260] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0261] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acidAttorney Docket No. 63340.32WO01sequence of a VL CDR2 set forth in Table 2 comprising 1 , 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0262] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0263] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2; the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2.

[0264] In some embodiments, the amino acid sequence of VH CDR1 consists of the aminoAttorney Docket No. 63340.32WO01acid of a VH CDR 1 set forth in Table 2, comprising no more than 1 , 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0265] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%. 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises the amino acid sequence of a VL set forth in Table 2.

[0266] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or theAttorney Docket No. 63340.32WO01amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0267] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3. or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0268] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 comprises the aminoAttorney Docket No. 63340.32WO01acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0269] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0270] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and theAttorney Docket No. 63340.32WO01amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0271] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0272] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8.

[0273] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acidAttorney Docket No. 63340.32WO01variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0274] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 59, or the amino acid sequence set forth in SEQ ID NO: 59 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 60, or the amino acid sequence set forth in SEQ ID NO: 60 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0275] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 59, or the amino acid sequence set forth in SEQ ID NO: 59 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 60, or the amino acid sequence set forth in SEQ ID NO: 60 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence ofAttorney Docket No. 63340.32WO01VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0276] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 59; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO; 60; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0277] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 59 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 60 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).Attorney Docket No. 63340.32WO01

[0278] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 59, or the amino acid sequence set forth in SEQ ID NO: 59 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 60, or the amino acid sequence set forth in SEQ ID NO: 60 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0279] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 59, or the amino acid sequence set forth in SEQ ID NO: 59 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 60, or the amino acid sequence set forth in SEQ ID NO: 60 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acidAttorney Docket No. 63340.32WO01sequence set forth in SEQ ID NO: 8 comprising no more than 1 , 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0280] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 59; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 60; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5: the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8.

[0281] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 59 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 60 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0282] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 61, or the amino acid sequence set forth in SEQ ID NO: 61 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 62, or the amino acid sequence set forth in SEQ ID NO: 62 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6,Attorney Docket No. 63340.32WO01or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1 , 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0283] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 61, or the amino acid sequence set forth in SEQ ID NO: 61 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 62, or the amino acid sequence set forth in SEQ ID NO: 62 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO; 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8. or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0284] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 61; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 62; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.Attorney Docket No. 63340.32WO01

[0285] In some embodiments, the amino acid sequence of VH CDR 1 the amino acid sequence set forth in SEQ ID NO: 61 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.): the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 62 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0286] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 61, or the amino acid sequence set forth in SEQ ID NO: 61 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 62, or the amino acid sequence set forth in SEQ ID NO: 62 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0287] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 61, or the amino acid sequence set forth in SEQ ID NO:Attorney Docket No. 63340.32WO0161 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 62, or the amino acid sequence set forth in SEQ ID NO: 62 comprising no more than 1, 2, or 3 amino acid variations e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0288] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 61; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 62; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8.

[0289] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 61 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 62 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequenceAttorney Docket No. 63340.32WO01of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0290] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, or the amino acid sequence set forth in SEQ ID NO: 63 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, or the amino acid sequence set forth in SEQ ID NO: 64 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65, or the amino acid sequence set forth in SEQ ID NO: 65 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 66, or the amino acid sequence set forth in SEQ ID NO: 66 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises amino acid sequence GAS, or amino acid sequence amino acid sequence GAS comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0291] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, or the amino acid sequence set forth in SEQ ID NO: 63 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, or the amino acid sequence set forth in SEQ ID NO: 64 comprising no more than I, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65, or the amino acid sequence set forth in SEQ ID NO: 65 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 66, or the amino acid sequence set forth in SEQ ID NO: 66 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion,Attorney Docket No. 63340.32WO01addition, etc.); the amino acid sequence of VL CDR2 comprises amino acid sequence GAS, or amino acid sequence GAS comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0292] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 66; the amino acid sequence of VL CDR2 comprises amino acid sequence GAS; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0293] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 63 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 66 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises amino acid sequence GAS comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0294] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 63, or the amino acid sequence set forth in SEQ ID NO: 63 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 64, or the amino acid sequence set forth in SEQ ID NO: 64 comprising 1, 2, or 3 amino acid variationsAttorney Docket No. 63340.32WO01(e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 65, or the amino acid sequence set forth in SEQ ID NO: 65 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 66, or the amino acid sequence set forth in SEQ ID NO: 66 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of amino acid sequence GAS, or amino acid sequence GAS comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0295] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 63, or the amino acid sequence set forth in SEQ ID NO: 63 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 64, or the amino acid sequence set forth in SEQ ID NO: 64 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 65, or the amino acid sequence set forth in SEQ ID NO: 65 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 66, or the amino acid sequence set forth in SEQ ID NO: 66 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of amino acid sequence GAS, or amino acid sequence GAS comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0296] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 63; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 64; the amino acid sequence of VH CDR3 consistsAttorney Docket No. 63340.32WO01of the amino acid sequence set forth in SEQ ID NO: 65; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 66; the amino acid sequence of VL CDR2 consists of amino acid sequence GAS; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8.

[0297] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 63 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 64 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 65 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 66 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of amino acid sequence GAS comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0298] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%. 86%, 87%, 88%, 89%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 99%. or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%. 96%. 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In someAttorney Docket No. 63340.32WO01embodiments, the amino acid sequence of the VH comprises the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises the amino acid sequence set forth in SEQ ID NO: 10.

[0299] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 10.

[0300] In some embodiments, the CD 161 binding protein comprises a light chain (LC) and a heavy chain (HC).

[0301] In some embodiments, the amino acid sequence of the HC comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%,Attorney Docket No. 63340.32WO0195%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC comprises an amino acid sequence at least 95%, 96%, 97%. 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC comprises an amino acid sequence at least 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC comprises the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC comprises the amino acid sequence set forth in SEQ ID NO: 68.

[0302] In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC consists of an amino acid sequence at least 95%, 96%, 97%. 98%. 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68. In some embodiments, the amino acid sequence of the HC consists of the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC consists of the amino acid sequence set forth in SEQ ID NO: 68.5.3 Ig Constant Regions

[0303] In some embodiments, a CD161 binding protein described herein (e.g., an anti-CD161 antibody (e.g., described herein)) comprises one or more Ig (e.g., human Ig (hlg)) constant region (e.g., a CHI region, a hinge region, a CH2 region, a CH3 region, an Fc region, 1 CL, K CL).

[0304] In some embodiments, the CD 161 binding protein (e.g., described herein) (e.g., an antiAttorney Docket No. 63340.32WO01CD161 antibody (e.g., described herein)) comprises one or more Ig (e.g., hlg) heavy chain constant region (e.g., a CHI region, a hinge region, a CH2 region, a CH3 region, an Fc region). For example, a full-length antibody CD 161 binding protein comprises a CHI region, a hinge region, a CH2 region, and a CH3 region. Or for example, the CD161 binding protein may be fused to one or more Ig e.g., hlg) heavy chain constant regions (e.g., a CHI region, a hinge region, a CH2 region, a CH3 region, an Fc region).

[0305] In some embodiments, the Ig is a hlg. In some embodiments, the Ig is a human IgG (IgG). In some embodiments, the hlgG is hlgGl, hIgG2 (e.g., hIgG2a or hIgG2b), hIgG3, or hIgG4. In some embodiments, the hlgG is hlgGl or hIgG4. In some embodiments, the hlgG is hlgGl. In some embodiments, the hlgG is hIgG4.

[0306] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Fc region. In some embodiments, the Fc region is part of a full-length antibody. In some embodiments, the Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Fc region comprises or consists of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of an IgGl hinge region, an IgGl CH2 region, and an IgGl CH3 region. In some embodiments, the Fc region comprises or consists of an IgGl hinge region, an IgGl CH2 region, and an IgGl CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region. In some embodiments, the Fc region comprises or consists of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of an hlgG hinge region, a hlgG CH2 region, and an hlgG CH3 region. In some embodiments, the Fc region comprises or consists of a hlgG hinge region, an hlgG CH2 region, and a hlgG CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of an hlgGl hinge region, an hlgGl CH2 region, and an hlgGl CH3 region. In some embodiments, the Fc region comprises or consists of an hlgGl hinge region, an hlgGl CH2 region, and an hlgGl CH3 region. In some embodiments, the Fc region comprises or consists of at least a portion of a hIgG4 hinge region, anAttorney Docket No. 63340.32WO01h!gG4 CH2 region, and an h!gG4 CH3 region. In some embodiments, the Fc region comprises or consists of a hIgG4 hinge region, an hIgG4 CH2 region, and a hIgG4 CH3 region.

[0307] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises one or more Ig (e.g., hlg) light chain constant regions (e.g., CL, KCL). In some embodiments, the one or more Ig light chain constant regions are hlg. For example, a full-length antibody comprises two light chains each comprising a light chain constant region (e.g., CL, KCL).

[0308] The amino acid sequence of exemplary reference IgGl and IgG4 heavy chain constant regions and Ig light chain constant regions, which can be incorporated in one or more of the embodiments described herein (e.g., CD161 binding proteins, conjugates, and fusion proteins), is provided in Table 3.Table 3. The Amino Acid Sequence of Exemplary hlg heavy and light chain constant regions & components.Attorney Docket No. 63340.32WO01Attorney Docket No. 63340.32WO01Attorney Docket No. 63340.32WO01

[0309] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, comprising or consisting of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD 161 binding protein (e.g., described herein)Attorney Docket No. 63340.32WO01(e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, comprising or consisting of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3. comprising or consisting of about no more than 1, 2, 3, 4, 5, 6, 7. 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions).

[0310] In some embodiments, the CD 161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, and further comprises 1 or more but less than 15% (less than 12%, less than 10%. less than 8%), amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, comprising or consisting of at least about 1, 2, 3. 4, 5, 6. 7, 8, 9, 10. or more amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, comprising or consisting of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising an amino acid sequence set forth in Table 3, comprising or consisting of about no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions.

[0311] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising at least about 1, 2, 3, 4, 5, 6, 7,Attorney Docket No. 63340.32WO018, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising or consisting about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising or consisting of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., amino acid substitutions, deletions, or additions).

[0312] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%). amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising or consisting at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising or consisting about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig constant region comprising the amino acid sequence of any one or more of SEQ ID NOS: 11-38, comprising or consisting of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions.5.3.1 Ig Effector Function

[0313] In some embodiments, the CD161 binding protein (e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises an Ig (e.g., hlg) Fc region. In someAttorney Docket No. 63340.32WO01embodiments, the Ig Fc region is part of a full-length antibody. Tn some embodiments, the Tg Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Ig Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgGl hinge region, an IgGl CH2 region, and an IgGl CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgGl hinge region, an IgGl CH2 region, and an IgGl CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region.

[0314] In some embodiments, the CD161 binding protein e.g., described herein) (e.g., an anti-CD161 antibody (e.g., described herein)) comprises a hlg Fc region. In some embodiments, the hlg Fc region is part of a full-length antibody. In some embodiments, the hlg Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the hlg Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the hlg Fc region comprises or consists of at least a portion of an hlgG hinge region, an hlgG CH2 region, and an hlgG CH3 region. In some embodiments, the hlg Fc region comprises or consists of an hlgG hinge region, an hlgG CH2 region, and an hlgG CH3 region. In some embodiments, the hlg Fc region comprises or consists of at least a portion of a hlgGl hinge region, a hlgGl CH2 region, and a hlgGl CH3 region. In some embodiments, the Ig Fc region comprises or consists of a hlgGl hinge region, a hlgGl CH2 region, and a hlgGl CH3 region. In some embodiments, the hlg Fc region comprises or consists of at least a portion of a hIgG4 hinge region, a hIgG4 CH2 region, and a hIgG4 CH3 region. In some embodiments, the hlg Fc region comprises or consists of a hIgG4 hinge region, a hIgG4 CH2 region, and a hIgG4 CH3 region.

[0315] In some embodiments, the Ig (e.g., hlg) Fc region exhibits an alteration (e.g., enhancement) in one or more Fc effector function relative to a reference (e.g., wild type) Ig Fc region. Exemplary Ig Fc effector functions include, but are not limited to, antibody dependentAttorney Docket No. 63340.32WO01cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), complement dependent cytotoxicity (CDC), binding affinity to Clq, and binding affinity to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa. and / or Fey R II lb (e.g., FcyRI, FcyRIIa, and / or FcyRIIIa))).

[0316] Standard in vitro and / or in vivo assays known in the art can be conducted to evaluate Ig (e.g., hlg) Fc effector function, including, any one or more of ADCC. CDC, ADCP. Fc receptor (e.g., Fey receptor) binding affinity, and Clq binding affinity.

[0317] For example, ADCC activity can be assessed utilizing standard (radioactive and nonradioactive) methods known in the art (see, e.g., W02006 / 082515, W02012 / 130831), the entire contents of each of which is incorporated by reference herein for all purposes). For example, ADCC activity can be assessed using a chromium-5 (51Cr) assay. Briefly, 51Cr is pre-loaded into target cells expressing CD20, NK cells are added to the culture, and radioactivity in the cell culture supernatant is assessed (indicative of lysis of the target cells by the NK cells). Similar nonradioactive assays can also be utilized that employ a similar method, but the target cells are pre-loaded with fluorescent dyes, such as calcein-AM, CFSE, BCECF, or lanthanide fluorophore (Europium), see, e.g., Parekh, Bhavin S et al. “Development and validation of an antibodydependent cell-mediated cytotoxicity-reporter gene assay.” mAbs vol. 4,3 (2012): 310-8. Doi: 10.4161 / mabs.19873, the entire contents of which is incorporated by reference herein for all purposes. Exemplary commercially available non-radioactive assays include, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (Cell Technology, Inc. Mountain View, Calif.; and CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, Wis.). Additional non-limiting examples of in vitro assays that can be used to assess ADCC activity of a CD161 binding protein (e.g., an anti-CD161 antibody) described herein include those described in US5500362; US5821337; Hellstrom, I., et al.. Proc. Nat’l Acad. Sci. USA 83 (1986) 7059-7063; Hellstrom, I., et al., Proc. Nat’l Acad. Sci. USA 82 (1985) 1499-1502; and Bruggemann, M„ et al., J. Exp. Med. 166 (1987) 1351-1361, the entire contents of each of which is incorporated by reference herein. Alternatively, or additionally, ADCC activity of a CD161 binding protein (e.g., an anti-CD161 antibody) described herein may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes, et al., Proc. Nat’l Acad. Sci. USA 95 (1998) 652-656, the entire contents of which is incorporated by reference herein for all purposes.

[0318] Clq binding assays can be utilized to assess the ability of a CD161 binding proteinAttorney Docket No. 63340.32WO01(e.g., an anti-CD161 antibody) described herein to bind Clq (or bind with less affinity than a reference a CD161 binding protein (e.g., an anti-CD161 antibody)) and hence lack (or have decreased) CDC activity. The binding of a CD161 binding protein (e.g., an anti-CD161 antibody) described herein to Clq can be determined by a variety of in vitro assays (e.g., biochemical or immunological based assays) known in the art for determining Fc-Clq interactions, including e.g., equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)), or kinetic methods (e.g., surface plasmon resonance (SPR) analysis), and other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods may utilize a label on one or more of the components being examined and / or employ a variety of detection methods including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinities and kinetics can be found in e.g., Paul, W. E., ed., Fundamental Immunology, 4th Ed., Lippincott-Raven, Philadelphia (1999), the entire contents of which is incorporated by reference herein. For example, see. e.g., Clq and C3c binding ELISAs described in W02006 / 029879 and W02005 / 100402, the entire contents of each of which is incorporated by reference herein for all purposes. Additional CDC activity assays include those described in e.g., Gazzano-Santoro, et al., J. Immunol. Methods 202 (1996) 163; Cragg, M. S., et al., Blood 101 (2003) 1045-1052; and Cragg, M. S., and Glennie, M. J., Blood 103 (2004) 2738-2743). the entire contents of each of which is incorporated by reference herein for all purposes.

[0319] ADCP activity can be measured by in vitro or in vivo methods known in the art and also commercially available assays (see, e.g., van de Donk NW, Moreau P, Plesner T, et al. “Clinical efficacy and management of monoclonal antibodies targeting CD38 and SLAMF7 in multiple myeloma,” Blood, 127(6):681-695 (2016), the entire contents of each of which is incorporated by reference herein for all purposes). For example, a primary cell based ADCP assay can be used in which fresh human peripheral blood mononuclear cells (PBMCs) are isolated, monocytes isolated and differentiated in culture to macrophages using standard procedures. The macrophages are fluorescently labeled added to cultures containing fluorescently labeled target cells expressing CD20 and a CD161 binding protein (e.g., an anti-CD161 antibody) described herein. Phagocytosis events can be analyzed using FACS screening and / or microscopy. A modified reporter version of the above described assay can also be used that employs an engineeredAttorney Docket No. 63340.32WO01cell line that stably expresses FcyRIIa (CD32a) as the effector cell line (e.g., an engineered T cell line, e.g., THP-1), removing the requirement for primary cells. Exemplary ADCP assays are described in e.g., Ackerman, M. E. et al. A robust, high-throughput assay to determine the phagocytic activity of clinical antibody samples. J. Immunol. Methods 366, 8-19 (2011); and Mcandrew, E. G. et al. Determining the phagocytic activity of clinical antibody samples. J. Vis. Exp. 3588 (2011). Doi: 10.3791 / 3588; the entire contents of each of which is incorporated by reference herein.

[0320] Binding of an Ig (e.g., hlg) (e.g., an antibody comprising the foregoing) to an Ig Fc receptor can be determined by a variety of in vitro assays (e.g., biochemical or immunological based assays) known in the art for determining Fc-Fc receptor interactions, i.e., specific binding of an Fc region to an Fc receptor. Common assays include equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)), or kinetic methods (e.g., surface plasmon resonance (SPR) analysis), and other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods may utilize a label on one or more of the components being examined and / or employ a variety of detection methods including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinities and kinetics can be found in e.g., Paul, W. E., ed., Fundamental Immunology, 4” Ed., Lippincott-Raven, Philadelphia (1999), the entire contents of which is incorporated by reference herein for all purposes.5.3.1.1 Enhanced Ig Effector Function

[0321] In some embodiments, the Ig Fc region of the CD161 binding protein (e.g., an antiCD 161 antibody) exhibits an enhancement (e.g., an increase) in one or more Fc effector function (e.g., relative to a reference (e.g., wild type) Ig Fc region). Exemplary Ig Fc effector functions include, but are not limited to, ADCC, ADCP, CDC, binding affinity to Clq, and binding affinity to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb (e.g., FcyRI, FcyRIIa, and / or FcyRIIIa))).

[0322] In some embodiments, the Ig Fc region of the CD161 binding protein (e.g., an anti-CD161 antibody) is modified (e.g., comprises one or more variation (e.g., one or more amino acid substitution, deletion, addition, etc.); altered glycosylation (e.g., afucosylation))) (referred to herein as a “modified Ig Fc”). In some embodiments, the modification (e.g., the variation (e.g.,Attorney Docket No. 63340.32WO01one or more amino acid substitution, deletion, addition, etc.); altered glycosylation (e.g., afucosylation))) enhances (e.g., increases) one or more Fc effector function, relative to a reference Ig Fc that does not comprise the modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))).

[0323] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) ADCC compared to a reference CD161 binding protein (e.g., an anti-CD161 antibody) that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein comprises a modified Ig Fc and exhibits enhanced (e.g., increased) CDC compared to a reference CD161 binding protein (e.g., an anti-CD161 antibody) that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) ADCP compared to a reference CD161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) binding affinity to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb (e.g., FcyRI, FcyRIIa, and / or FcyRIIIa))) compared to a reference CD161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) binding affinity to FcyRI, FcyRIIa. and / or FcyRIIIa compared to a reference CD 161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) binding affinity to FcyRI compared to a reference CD 161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acidAttorney Docket No. 63340.32WO01substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the a CD161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) to Fc RIIa compared to a reference CD 161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD161 binding protein comprises a modified Ig Fc and exhibits enhanced (e.g., increased) binding affinity to FcyRIIIa compared to a reference CD161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))). In some embodiments, the CD 161 binding protein (e.g., an anti-CD161 antibody) comprises a modified Ig Fc and exhibits enhanced (e.g., increased) binding affinity to Clq compared to a reference CD161 binding protein that does not comprise the Ig Fc modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation (e.g., afucosylation))).

[0324] Amino acid substitutions that enhance (e.g., increase) one or more Ig Fc effector function are known in the art. See for example, Liu R, Oldham RJ, Teal E, Beers SA, Cragg MS. Fc-Engineering for Modulated Effector Functions-Improving Antibodies for Cancer Treatment. Antibodies (Basel). 2020;9(4):64. Published 2020 Nov 17. doi:10.3390 / antib9040064; van der Horst HJ, Nijhof IS, Mutis T, Chamuleau MED. Fc-Engineered Antibodies with Enhanced Fc-Effector Function for the Treatment of B-Cell Malignancies. Cancers (Basel). 2020;12(10):3041. Published 2020 Oct 19. Doi:10.3390 / cancersl2103041; and Saunders Kevin, “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life,” Frontiers in Immunology, vlO (June 7, 2019) DOI=10.3389 / fimmu.2019.01296, the full contents of each of which is incorporated by reference herein for all purposes.

[0325] Table 4 below, provides exemplary amino acid substitutions (and combinations thereof) and alterations in glycosylation (e.g., afucosylation) that can be utilized to increase one or more Ig Fc effector function. Amino acids in Table 4 are numbered according to the EU numbering scheme. The effects on effector function set forth in Table 4 are exemplary only and not intended to be limiting. The amino acid substitutions set forth in Table 4 are with reference to an IgGl Fc region (except where noted). However, a person of ordinary skill in the could identify the corresponding amino acid in a non-IgGl Fc region, for example in an IgG2 or IgG4 Fc region,Attorney Docket No. 63340.32WO01should the base amino acid be different between the IgGl and non-IgGl Fc region.Table 4. Exemplary Ig Fc Variations and Glycoengineering to Increase Effector Function.

[0326] In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD161 binding protein (e.g., an anti-CD161 antibody) comprises any one or more of the amino acid substitutions set forth in Table 4 (i.e., any one or more amino acid substitution set forth in any set of amino acid substitutions set forth in Table 4). In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD161 binding protein (e.g., an anti-CD161 antibody) comprises any one or more of the sets of amino acid substitutions set forth in Table 4. In some embodiments, the Ig Fc (e.g., IgGl Fc) regionAttorney Docket No. 63340.32WO01of a CD161 binding protein (e.g., an anti-CD161 antibody) comprises any one or more of the glycosylation changes set forth in Table 4.

[0327] In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD 161 binding protein (e.g., an anti-CD161 antibody) comprises an amino acid substitution at any one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of amino acid positions S298, E333, K334, S239, 1332, P247, A339, A330. G236, F243. R292, Y300, V305. P396, L235. F243, R292. Y300, P396, F243, R292, Y300, V305, P396, K326, E333, S267E, H268, S324, S298, E333, K334, L234, L235, G236, S239, H268, D270, S298 D270, K326, A330, and / or K334. In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD 161 binding protein (e.g., an anti-CD161 antibody) comprises an amino acid substitution at from about 1-10 (e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid positions S298, E333, K334, S239.1332. P247, A339, A330, G236, F243. R292, Y300, V305, P396, L235, F243, R292, Y300, P396, F243, R292, Y300, V305, P396, K326, E333, S267E, H268. S324, S298, E333, K334, L234, L235, G236. S239, H268, D270, S298 D270, K326, A330, and / or K334.

[0328] In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD161 binding protein (e.g., an anti-CD161 antibody) comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of the following amino acid substitutions S298A, E333A, K334A, S239D, I332E, P247I, A339Q, A330L, G236A, F243L, R292P, Y300L, V305I, P396L, L235V, F243L, R292P, Y300L, P396L, F243L, R292P, Y300L, V305I, P396L, K326W, E333S, S267E, H268E, S324T, S298A, E333A, K334A, L234Y, L235Q, G236W, S239M, H268D, D270E, S298A D270E, K326D, A330M, and / or K334E.

[0329] In some embodiments, the Ig Fc (e.g., IgGl Fc) region of a CD 161 binding protein (e.g., an anti-CD161 antibody) comprises from about 1-10 (e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid substitutions S298A, E333A, K334A, S239D, I332E, P247I, A339Q, A330L, G236A, F243L. R292P, Y300L. V305I. P396L, L235V. F243L, R292P, Y300L, P396L, F243L, R292P, Y300L, V305I, P396L, K326W, E333S, S267E, H268E, S324T, S298A, E333A, K334A, L234Y, L235Q, G236W, S239M, H268D. D270E, S298A D270E, K326D, A330M, and / or K334E.

[0330] In some embodiments, the Ig Fc region of a CD 161 binding protein (e.g., an antiCD 161 antibody) comprises an IgGl Fc region comprising one or more amino acid variation relative to a reference IgGl Fc region.Attorney Docket No. 63340.32WO01

[0331] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S298, E333, K334, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S298A, E333A, and / or K334A, EU numbering according to Kabat.

[0332] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions S239 and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions S239D and / or I332E, EU numbering according to Kabat.

[0333] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions P247 and / or A339. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions P247I and / or A339Q, EU numbering according to Kabat.

[0334] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S239, A330, and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S239D, A330L, and / or I332E, EU numbering according to Kabat.

[0335] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions G236, S239, and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions G236A, S239D, and / or I332E, EU numbering according to Kabat.

[0336] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, 4, or 5) of amino acid positions F243, R292, Y300. V305, and / or P396. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, 3, 4, or 5) of the following amino acid substitutions F243L, R292P, Y300L, V305I, and / or P396L, EU numbering according to Kabat.

[0337] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, 4, or 5) of amino acid positions L235, F243, R292, Y300, and P396. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or moreAttorney Docket No. 63340.32WO01(e.g., 1, 2, 3, 4, or 5) of the following amino acid substitutions L235V, F243L, R292P, Y300L, and / or P396L, EU numbering according to Kabat.

[0338] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of amino acid positions L234, L235, G236, S239, H268, D270, and / or S298, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, 3, 4, 5. 6, or 7) of the following amino acid substitutions L234Y, L235Q, G236W, S239M, H268D, D270E, and / or S298A, EU numbering according to Kabat.

[0339] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, or 4) of amino acid positions D270, K326, A330, and / or K334, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, 3, or 4) of the following amino acid substitutions D270E, K326D, A330M, and / or K334E, EU numbering according to Kabat.

[0340] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, 4, or 5) of amino acid positions F243, R292, Y300. V305, and / or P396. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, 3, 4, or 5) of the following amino acid substitutions F243L, R292P, Y300L, V305I, and / or P396L, EU numbering according to Kabat.

[0341] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S239, 1332, and / or A330, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S239D, I332E, and / or A330L, EU numbering according to Kabat.

[0342] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, or 4) of amino acid positions S239, 1332, A330, and / or G236, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., I. 2, 3, or 4) of the following amino acid substitutions S239D, I332E, A330L and / or G236A, EU numbering according to Kabat.

[0343] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S239, 1332, and / or G326, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S239D, I332E, and / or G326A, EU numbering according toAttorney Docket No. 63340.32WO01Kabat.

[0344] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position G326, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a G326A amino acid substitution, EU numbering according to Kabat.

[0345] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions G236. S239, and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions G236A, S239D, and / or I332E, EU numbering according to Kabat.

[0346] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions S239 and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions S239D and / or I332E, EU numbering according to Kabat.

[0347] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions K326 and / or E333, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions K326W and / or E333S, EU numbering according to Kabat.

[0348] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S267, H268, and / or S324, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S267E, H268E, and / or S324T, EU numbering according to Kabat.

[0349] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S298, E333, and / or K334, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., I. 2, or 3) of the following amino acid substitutions S298A, E333A, and / or K334A, EU numbering according to Kabat.

[0350] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions S239 and / or 1332, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions S239D and / or I332E, EU numbering according to Kabat.Attorney Docket No. 63340.32WO01

[0351] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions P247 and / or A339, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1 or 2) of the following amino acid substitutions P247I and / or A339Q, EU numbering according to Kabat.

[0352] In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., an antiCD 161 antibody) comprises one or more changes to the glycosylation.

[0353] In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., an anti-CD161 antibody) is afucosylated. In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., an anti-CD161 antibody) is afucosylated and a CD161 binding protein (e.g., an anti-CD161 antibody) comprising the afucosylated Ig Fc exhibits enhanced (e.g., increased) ADCC compared to a reference CD 161 binding protein that is not afucosylated. Methods of producing afucosylated antibodies are known in the art. See, e.g., Pereira, Natasha A et al. “The "less-is-more" in therapeutic antibodies: Afucosylated anti-cancer antibodies with enhanced antibodydependent cellular cytotoxicity.” mAbs vol. 10,5 (2018): 693-711. doi:10.1080 / 19420862.2018.1466767, the entire contents of which is incorporated herein by reference for all purposes.5.3.1.2 Reduced Ig Effector F unction

[0354] In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., described herein) exhibits a decrease in one or more Fc effector function relative to a reference (e.g., wild type) Ig Fc region. Exemplary Ig Fc effector functions include, but are not limited to, ADCC, ADCP, CDC, binding affinity to Clq, and binding affinity to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb (e.g., FcyRI, FcyRIIa, and / or FcyRIIIa))).

[0355] In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., described herein) is modified (e.g., comprises one or more variation (e.g., one or more amino acid substitution, deletion, addition, etc.); altered glycosylation)) (referred to herein as a “modified Ig Fc”). In some embodiments, the one or more variation (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)) decreases or abolishes one or more Fc effector function, relative to a reference Ig Fc that does not comprise the modification (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)).Attorney Docket No. 63340.32WO01

[0356] In some embodiments, the modified Ig Fc fusion protein exhibits no detectable or decreased ADCC compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits no detectable or decreased CDC compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits no detectable or decreased ADCP compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to one or more human Fc receptor {e.g., an Fey receptor {e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb {e.g., FcyRI, FcyRIIa, and / or FcyRIIIa))) compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to FcyRI, FcyRIIa, and / or FcyRIIIa compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to FcyRI compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to FcyRIIa compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to FcyRIIIa compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the one or more variation {e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)). In some embodiments, the modified Ig Fc fusion protein exhibits decreased or no binding affinity to Clq compared to a reference fusion protein that does not comprise the Ig Fc modification {e.g., the oneAttorney Docket No. 63340.32WO01or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.; the altered glycosylation)).

[0357] Amino acid substitutions that decrease or abolish one or more Ig Fc effector function are known in the art. See for example, Saunders Kevin, “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life,” Frontiers in Immunology, vlO (June 7, 2019) DOI=10.3389 / fimmu.2019.01296, the full contents of which is incorporated by reference herein for all purposes, see more particularly for example, e.g., Table 3 of Saunders.

[0358] Table 5 below, provides exemplary amino acid substitutions (and combinations thereof) that can be utilized to increase one or more Fc effector function. Amino acids in Table 5 are numbered according to the EU numbering scheme. The effects on effector function set forth in Table 5 are exemplary only and not intended to be limiting. The amino acid substitutions set forth in Table 5 (except where noted) are with reference to an IgGl Fc region. However, a person of ordinary skill in the could identify the corresponding amino acid in a non-IgGl Fc region, for example in an IgG2 or IgG4 Fc region, should the base amino acid be different between the IgGl and non-IgGl Fc region.Table 5. Exemplary Ig Fc Variations and Glycoengineering to Decrease Effector Function.Attorney Docket No. 63340.32WO01

[0359] In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD161 binding protein {e.g., described herein) comprises any one or more of the amino acid substitutions set forth in Table 5 (set forth in any set). In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD161 binding protein {e.g., described herein) comprises any one or more of the sets of amino acid substitutions set forth in Table 5. In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD161 binding protein {e.g., described herein) comprises any one or more of the glycosylation changes set forth in Table 5.

[0360] In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD 161 binding protein {e.g., described herein) comprises an amino acid substitution at any one or more {e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of amino acid positions L234, L235, P329, P331, D265, G237, E318, E233. G236, L328, D270. K322, V264, F241, and / or N297. In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD 161 binding protein {e.g., described herein) comprises an amino acid substitution at from about 1-10 {e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid positions L234, L235, P329, P331, D265, G237, E318, E233, G236, L328, D270, K322, V264, F241, and / or N297.

[0361] In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD 161 binding protein {e.g., described herein) comprises one or more {e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of the following amino acid substitutions L234A, L234G, L235F, L235E, L235A, P329G, P329A, P331S, D265A, G237A, E318A, E233P, G236R, L328R, D270A, K322A, V264A, F241A, N297A, N297G, and / or N297Q. In some embodiments, the Ig Fc {e.g., IgGl Fc) region of a CD161 binding protein {e.g., described herein) comprises from about 1-10 {e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid substitutions L234A, L234G, L235F, L235E, L235A, P329G, P329A, P331S, D265A, G237A, E318A, E233P, G236R, L328R, D270A, K322A, V264A, F241A, N297A, N297G. or N297Q.

[0362] In some embodiments, the Ig Fc region of a CD161 binding protein {e.g., described herein) comprises an IgGl Fc region comprising one or more amino acid variation.Attorney Docket No. 63340.32WO01

[0363] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position L234, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a L234A or L234G amino acid substitution, EU numbering according to Kabat.

[0364] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position L235. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a L235A, L235G. L235E, or L235F amino acid substitution, EU numbering according to Kabat.

[0365] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position P329, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a P329A or P329G amino acid substitution, EU numbering according to Kabat.

[0366] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions L234 and / or L235, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A and / or L235A, EU numbering according to Kabat.

[0367] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions L234, L235, and / or P329, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A, L235A, and / or P329G, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A, L235A, and / or P329A, EU numbering according to Kabat.

[0368] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions P331, L234, and / or L235, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions P331S. L234G. and / or L235F, EU numbering according to Kabat.

[0369] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position D265, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a D235A amino acid substitution, EU numbering according to Kabat.Attorney Docket No. 63340.32WO01

[0370] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position G237, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a G237A amino acid substitution, EU numbering according to Kabat.

[0371] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position E318. EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a E318A amino acid substitution, EU numbering according to Kabat.

[0372] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position E233, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a E233P amino acid substitution, EU numbering according to Kabat.

[0373] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position D270, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a D270A amino acid substitution, EU numbering according to Kabat.

[0374] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position K322, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a K322A amino acid substitution, EU numbering according to Kabat.

[0375] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position P331, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a P331A amino acid substitution, EU numbering according to Kabat.

[0376] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position F241, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a F241A amino acid substitution, EU numbering according to Kabat.

[0377] In some embodiments, the IgGl Fc region comprises an amino acid substitution at amino acid position N297, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises a N297A, N297G, or N297Q amino acid substitution, EU numbering according to Kabat.

[0378] In some embodiments, the IgGl Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions G236 and / or L328, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions G236R and / or L328R, EU numbering according to Kabat.Attorney Docket No. 63340.32WO01

[0379] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at amino acid position S228, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises an S228P amino acid substitution, EU numbering according to Kabat.

[0380] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at amino acid position F234, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises an F234A amino acid substitution, EU numbering according to Kabat.

[0381] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at amino acid position L235A, EU numbering according to Kabat. In some embodiments, the IgGl Fc region comprises an L235A, L235G, or L235E amino acid substitution, EU numbering according to Kabat.

[0382] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at one or more {e.g., 1 or 2) of amino acid positions S228 and / or L235, EU numbering according to Kabat. In some embodiments, the IgG4 Fc region comprises one or more {e.g., 1, 2, or 3) of the following amino acid substitutions S228P and / or L235E, EU numbering according to Kabat.

[0383] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at one or more {e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the IgG4 Fc region comprises one or more {e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235A, EU numbering according to Kabat.

[0384] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at one or more {e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the IgG4 Fc region comprises one or more {e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235G, EU numbering according to Kabat.

[0385] In some embodiments, the IgG4 Fc region comprises an amino acid substitution at one or more e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the IgG4 Fc region comprises one or more {e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235E, EU numbering according to Kabat.

[0386] In some embodiments, the IgG2 Fc region comprises an amino acid substitution at one or more {e.g., 1, 2, 3, or 4) of amino acid positions H268, V309, A330, and / or P331, EU numberingAttorney Docket No. 63340.32WO01according to Kabat. In some embodiments, the IgG2 Fc region comprises one or more (e.g., 1, 2, 3, or 4) of the following amino acid substitutions H268Q, V309L, A330S, and / or P331S, EU numbering according to Kabat.

[0387] In some embodiments, the Ig Fc region of a CD161 binding protein (e.g., described herein) comprises one or more changes to the glycosylation. In some embodiments, the Ig Fc region of a CD 161 binding protein (e.g., described herein) has increased high mannose glycosylation.5.3.2 Promotion of Heterodimerization

[0388] As described herein, in some embodiments, the antibody comprises a first and second Fc region. In some embodiments, the first Ig Fc region and the second Ig Fc region each comprise one or more amino acid modifications relative to each other to promote heterodimerization. IgG derived heterodimeric formats can be generated by methods known in the art, e.g., by forced heavy chain heterodimerization. Forced heavy chain heterodimerization can be obtained using known methods in the art, e.g., knob-in-hole or strand exchange engineered domains (SEED), see, e.g., Ji-Hee et al., “Immunoglobulin Fc Heterodimer Platform Technology: From Design to Applications in Therapeutic Antibodies and Proteins” Frontiers in Immunology, v7 (article 394) (2016) DOI=10.3389 / fimmu.2016.00394 (hereinafter “Ji-Hee 2016”). the entire contents of which is incorporated by reference herein for all purposes.

[0389] In some embodiments, an interface of the first and the second Ig Fc regions is varied, e.g., introduction of an amino acid substitution, to increase heterodimerization, e.g., relative to a non-modified interface, e.g., a naturally occurring interface. For example, dimerization of the first and second Ig Fc regions can be enhanced by providing an Ig Fc interface of a first and a second Fc region with one or more of: a paired protuberance-cavity (“knob-in-hole”), an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms, e.g., relative to a non-modified interface.

[0390] Knob-in-Hole amino acid pairing modifications are known in the art, and described in e.g., US5731116; US7476724; Ji-Hee 2016; and Ridgway, J. “'Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization” etal. Prot. Engineering 9(7): 617-621 (1996), the full contents of each of which is incorporated by reference herein. Generally, Knob-in-Hole comprises 1) introducing one or more amino acid substitutions in the CH3 domain of one orAttorney Docket No. 63340.32WO01both of the first and second subject Ig Fc regions to promote heterodimerization; and 2) combining the modified Ig Fc regions under conditions that promote heterodimerization. “Knobs” are typically created by substituting a small amino acid in a parental Ig Fc region with a larger amino acid (e.g., T366Y or T366W); “holes” are created by substituting a larger residue in a parental Ig Fc region with a smaller amino acid (e.g., Y407T, T366S, 11368A, or Y407V). Exemplary Knobin-Hole mutations include S354C, T366W in the “knob” Ig Fc region and Y349C, T366S, L368A, Y407V in the “hole” Ig Fc region. Other exemplary Knob-in-Hole mutations, which can be incorporated into any one or more of the embodiments, are provided in Table 6, with additional exemplary optional stabilizing Ig Fc cysteine mutations.Table 6. Exemplary Knob-in-hole and Stabilizing Cysteine Modifications.

[0391] As described herein, in some embodiments, the antibody comprises a first Fc region and a second Ig Fc region.

[0392] In some embodiments, the amino acid sequence of the first Fc region comprises a T366W amino acid substitution, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, and Y407V, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3). In some embodiments, the amino acid sequence of the first hlg further comprises a S354C amino acid substitution, EU numbering according to Kabat; and the amino acid sequence of the second Fc region comprises a Y349C amino acid substitution, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0393] In some embodiments, the amino acid sequence of the first Fc region comprises each of the following amino acid substitutions: T366W and a S354C, EU numbering according toAttorney Docket No. 63340.32WO01Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, Y407V, and Y349C, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0394] In some embodiments, the amino acid sequence of the second Fc region comprises a T366W amino acid substitution, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, and Y407V, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3). In some embodiments, the amino acid sequence of the second hlg further comprises a S354C amino acid substitution, EU numbering according to Kabat; and the amino acid sequence of the second Fc region comprises a Y349C amino acid substitution, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0395] In some embodiments, the amino acid sequence of the second Fc region comprises each of the following amino acid substitutions: T366W and S354C, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, Y407V, and Y349C, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0396] In some embodiments, the amino acid sequence of the first Ig Fc region comprises a W amino acid at position T366, EU numbering according to Kabat; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat.

[0397] In some embodiments, the amino acid sequence of the first Ig Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat.5.4 Conjugates and Fusion ProteinsAttorney Docket No. 63340.32WO01

[0398] In some embodiments, a CD161 binding protein described herein (e.g., an anti-CDl 61 antibody) is conjugated or fused to one or more heterologous moiety. As such, further provided herein are fusion proteins comprising a CD 161 binding protein described herein (e.g., an antiCD 161 antibody) fused (directly or indirectly (via a linker)) to one or more heterologous protein. Further provided herein are conjugates comprising a CD161 binding protein described herein (e.g., an anti-CD161 antibody) conjugated (directly or indirectly (via a linker)) to one or more heterologous moiety.

[0399] In some embodiments, the heterologous moiety is a protein, peptide, polynucleotide (e.g., DNA, RNA, DNA / RNA hybrid), small molecule, carbohydrate, lipid, or synthetic polymer.

[0400] In some embodiments, the heterologous protein comprises an IgG (e.g., IgGl, IgG2 (e.g., IgG2a or IgG2b), IgG3, IgG4), IgE, IgM, IgD, or IgA (e.g., IgAl or IgA2) antibody. In some embodiments, the heterologous polypeptide is an antibody (e.g., a full-length antibody, scFv, Fab, F(ab')2, Fab', Fv, single domain antibody (e.g., a VHH), scFv-Fc, Fab-Fc, and / or single domain antibody-Fc (e.g., VHH-Fc)). In some embodiments, the heterologous protein comprises one or more of a monoclonal antibody, monospecific antibody, multispecific antibody, human antibody, humanized antibody, chimeric antibody, and / or murine antibody, or a functional fragment or functional variant of any of the foregoing. In some embodiments, the antibody is a chimeric antigen receptor (e.g., described herein). In some embodiments, the antibody is a T-cell engager (e.g., described herein).

[0401] In some embodiments, the heterologous moiety is a therapeutic agent. In some embodiments, the heterologous moiety is a chemotherapeutic agent, a cytotoxic agent, an anticancer agent, or a radioactive isotope, or any combination thereof. In some embodiments, the heterologous moiety is a targeting moiety. In some embodiments, the heterologous moiety is a detectable moiety. In some embodiments, the heterologous moiety is a diagnostic moiety. In some embodiments, the heterologous moiety is a fluorescent moiety. Fluorescent or chemiluminescent labels include fluorophores such as rare earth chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanate, phycoerythrin, phycocyanin, allophycocyanin, o-phthaladehyde, fluorescamine, 152Eu, dansyl, umbelliferone, luciferin, luminal label, isoluminal label, an aromatic acridinium ester label, an imidazole label, an acridimium salt label, an oxalate ester label, an aequorin label, 2,3-dihydrophthalazinediones, biotin / avidin. spin labels and stable free radicals.Attorney Docket No. 63340.32WO01

[0402] In some embodiments, one or more polypeptide of a CD161 binding protein (or one or more polypeptide thereof) or polypeptide (e.g., described herein) comprises a signal peptide operably connected to the N-terminus. Commonly utilized signal peptides are known in the art, for example, the native signal peptide of human interleukin 2 (hIL-2), human oncostatin M (hOSM), human chymotrypsinogen (hCTRBl), human trypsinogen 2 (hTRY2), and human insulin (hINS). A person of ordinary skill can determine the appropriate signal peptide using standard methodology known in the art.5.4.1 Chimeric Antigen Receptors

[0403] In some embodiments, a CD 161 binding protein described herein is part of a chimeric antigen receptor (CAR). In some embodiments, a CD161 binding protein described herein is the extracellular antigen-binding domain of a CAR. Standard CAR domains are known in art, including, e.g., transmembrane domains and intracellular signaling domains. See, e.g., W02024056809, W02023240064A1, and WO2023205148A1, WO2023133092A1, the entire contents of each of which is incorporated herein by reference for all purposes.

[0404] Exemplary transmembrane domains include, e.g., the alpha, beta or zeta chain of T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8 (for example, CD8 alpha, CD8 beta), CD9, CD 16, CD22, CD33, CD37, CD64, CD80. CD86. CD134, CD137, CD154. In some embodiments, a transmembrane domain may include at least the transmembrane region(s) of a costimulatory molecule, for example, MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, 0X40, CD2, CD7, CD27, CD28, CD30, CD40. CDS, ICAM-1, LFA-1 (CDUa / CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD 19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma. IL7R alpha, ITGA4, VLA1. CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CDlld, ITGAE, CD103, ITGAL, CDlla, LFA-1, ITGAM, CDllb, ITGAX, CDllc, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD 150, IPO-3), BLAME (SLAMF8), SELPLG (CD 162), LTBR, LAT, GADS. SLP-Attorney Docket No. 63340.32WO0176, PAG / Cbp, CD 19a, and a ligand that specifically binds with CD83. In some instances, the transmembrane domain can be attached to the extracellular region of the CAR, for example, the antigen-binding domain of the CAR, via a hinge, for example, a hinge from a human protein. For example, in some embodiments, the hinge can be a human Ig (immunoglobulin) hinge, for example, an IgG4 hinge, or a CD8a hinge.

[0405] Exemplary intracellular signaling domains include, e.g., the cytoplasmic sequences of the T cell receptor (TCR) and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any recombinant sequence that has the same functional capability. In some embodiments, the intracellular signaling domain comprises a primary signaling domain and one or more costimulatory signaling domain. Exemplary primary signaling domains, include, e.g., intracellular signaling domains of TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, CD278 (also known as “ICOS”), FccRI, DAP10, DAP12, CD32, and CD66d. Exemplary of proteins with costimulatory domains suitable for use in CAR described herein include, e.g., MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, 0X40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD1 la / CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT. HVEM (LIGHTR). KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD 19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD lid, ITGAE, CD 103, ITGAL, CDlla. LFA-1, ITGAM. CDllb. ITGAX, CDllc, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT AM, Ly9 (CD229), CD 160 (BY55). PSGL1. CD100 (SEMA4D), CD69. SLAMF6 (NTB-A. Lyl08), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD 162), LTBR, LAT, GADS, SLP-76, PAG / Cbp, CD 19a, and a ligand that specifically binds with CD83, and the like.5.4.2 T-Cell Engagers

[0406] In some embodiments, a CD 161 binding protein described herein is part of a T cell engager (i.e., an agent (e.g., antibody) that binds at one molecule (e.g., protein) expressed on theAttorney Docket No. 63340.32WO01surface of a T-cell). In some embodiments, the T-cell engager is multispecific (e.g., bispecific, trispecific, etc). In some embodiments, the T-cell engager is bispecific. In some embodiments, the T-cell engager is trispecific. In some embodiments, the T-cell engager binds to least one molecule (e.g., protein) expressed on the surface of a T cell and at least one molecule expressed on the surface of a non-T-cell. In some embodiments, the CD161 binding protein acts as the T-cell engager.5.4.3 Half-Life Extension Moieties

[0407] In some embodiments, the heterologous moiety (e.g., protein) is a half-life extension moiety e.g., protein). Various half-life extension moieties are known in the art. See, e.g., Ko S, Jo M, Jung ST. Recent Achievements and Challenges in Prolonging the Serum Half-Lives of Therapeutic IgG Antibodies Through Fc Engineering. BioDrugs. 2021 ;35(2): 147- 157. doi: 10.1007 / s40259-021-00471-0 (hereinafter “Ko 2021”); Bech, E. M., Pedersen, S. L., & Jensen, K. J. (2018). Chemical Strategies for Half-Life Extension of Biopharmaceuticals: Lipidation and Its Alternatives. A CS medicinal chemistry letters, 9(7), 577-580. https: / / doi.org / 10.1021 / acsmedchemlett.8b00226 (hereinafter “Bech 2018”); Mester S, Evers M, Meyer S, et al. Extended plasma half-life of albumin-binding domain fused human IgA upon pH-dependent albumin engagement of human FcRn in vitro and in vivo. MAhs. 2021 ; 13(1): 1893888. doi: 10.1080 / 19420862.2021.1893888 (hereinafter “Mester 2021”); Kontermann RE. Strategies for extended serum half-life of protein therapeutics. Curr Opin Biotechnol. 2011;22(6):868-876. doi: 10.1016 / j. copbio.2011.06.012 (hereinafter “Kontermann 2011”); Strohl W. R. (2015). Fusion Proteins for Half-Life Extension of Biologies as a Strategy to Make Biobetters. BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy, 29(4), 215-239. https: / / doi.org / 10.1007 / s40259-015-0133-6; Zaman R, Islam RA, Ibnat N, et al. Current strategies in extending half-lives of therapeutic proteins. J Control Release. 2019;301:176-189. doi:10.1016 / j.jconrel.2019.02.016: Chen C, Constantinou A, Chester KA, et al. Glycoengineering approach to half-life extension of recombinant biotherapeutics. Bioconjug Chem.2012;23(8): 1524-1533. doi:10.1021 / bc200624a; Gupta, Vijayalaxmi et al. “Protein PEGylation for cancer therapy: bench to bedside.” Journal of cell communication and signaling vol. 13,3 (2019): 319-330. doi:10.1007 / sl2079-018-0492-0; Martin Schlapschy, et al, PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically activeAttorney Docket No. 63340.32WO01proteins, Protein Engineering, Design and Selection, Volume 26, Issue 8, August 2013, Pages 489-501, https: / / doi.org / 10.1093 / protein / gzt023; Strohl, William R. “Fusion Proteins for Half-Life Extension of Biologies as a Strategy to Make Biobetters.” BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy vol. 29,4 (2015): 215-39. doi:10.1007 / s40259-015-0133-6; the entire contents of each of which are incorporated by reference herein for all purposes.

[0408] Exemplary half-life extension moieties include, but are not limited to, an immunoglobulin (e.g., human Ig (hlg), murine Ig (mlg)), a fragment of an Ig (e.g., hlg, mlg), an Ig (e.g., hlg, mlg) constant region, a fragment of an Ig (e.g., hlg, mlg) constant region, an Ig {e.g., hlg, mlg) Fc region, human transferrin, a human transferrin binding moiety {e.g., small molecule, lipid, protein, peptide, etc.), human serum albumin (HSA), a fragment of HSA, an HSA binding moiety {e.g., small molecule, lipid, protein, peptide, etc.) {e.g., an antibody, a Streptococcal protein G {see, e.g., Mester 2021), polyethylene glycol (PEG) (and polymers thereof) {e.g., pegylation), lipids, small molecules, carbohydrates {e.g., glycosylation, polysialic acid (polysialylation), hydroxyethyl starch (HES) (HESylation), heparosan (HEPylation)).

[0409] In some embodiments, the heterologous polypeptide is a half-life extension polypeptide. Exemplary half-life extension polypeptides include, but are not limited to, an Ig, a fragment of an Ig, one or more Ig heavy chain constant region, a fragment of an Ig constant region, an Ig Fc region, a hlg, a fragment of a hlg, one or more hlg heavy chain constant region, a fragment of a hlg constant region, a hlg Fc region, a mlg, a fragment of a mlg, one or more mlg heavy chain constant region, a fragment of a mlg constant region, a mlg Fc region, human transferrin, a fragment of human transferrin, a human transferrin binding protein {e.g., an antibody) HSA, and HSA binding proteins {e.g., an antibody, a Streptococcal protein G). In some embodiments, the half-life extension polypeptide comprises an Ig Fc region {e.g., hlg Fc region). In some embodiments, the Ig (e.g., hlg, mlg) Fc region of a fusion protein described herein comprises one or more amino acid variation (e.g., relative to a reference Ig {e.g., hlg, mlg) Fc region {e.g., a wildtype Ig (e.g., hlg, mlg) Fc region)) that enhances serum half-life of the fusion protein (e.g., relative to a reference Ig (e.g., hlg, mlg) Fc region (e.g., a wild-type Ig (e.g., hlg, mlg) Fc region)). See, e.g., § 5.3.

[0410] In some embodiments, half-life extension is mediated through one or more of lipidation, glycosylation, polysialylation, HESylation, HEPylation, and / or pegylation. In someAttorney Docket No. 63340.32WO01embodiments, half-life extension is mediated through one or more of lipidation, HESylation, HEPylation, and / or pegylation. In some embodiments, half-life extension is mediated through glycosylation. In some embodiments, half-life extension is mediated through polysialylation.

[0411] In some embodiments, the half-life extension moiety comprises one or more lipids. See, e.g., Bech 2018. In some embodiments, the half-life extension moiety comprises one or more post translational modifications (e.g., glycosylation, polysialylation, etc.).

[0412] In some embodiments, the half-life extension moiety e.g., protein) is altered (e.g., compared to a reference half-life extension moiety (e.g., protein)) to further enhance half-life of the fusion protein or conjugate. Various alterations to known half-life extension moieties (e.g., proteins) are known in the art. See, e.g., Ko 2021, Bech 2018, Mester 2021, and Kontermann 2011. Modifications include, e.g., amino acid variations (e.g., substitutions, additions, deletions) and post translational modifications (e.g., altered lipidation, glycosylation, polysialylation, HESylation, HEPylation, pegylation, etc.).

[0413] The immunoreceptor inhibitory protein described herein fused or conjugated to a halflife extending moiety or a half-life extending moiety can be evaluated for their pharmacokinetic properties utilizing standard in vivo methods known in the art. See, e.g., Avery, Lindsay B et al. “Utility of a human FcRn transgenic mouse model in drug discovery for early assessment and prediction of human pharmacokinetics of monoclonal antibodies.” mAbs vol. 8,6 (2016): 1064-78. doi:10.1080 / 19420862.2016.1193660; Conner, Christopher M et al. “A precisely humanized FCRN transgenic mouse for preclinical pharmacokinetics studies.” Biochemical pharmacology vol. 210 (2023): 115470. doi:10.1016 / j.bcp.2023.115470; and Kathryn Ball et al., PK and Biodistribution of Therapeutic Proteins. Drug Metabolism and Disposition June 1, 2022, 50 (6) 858-866; DOI: https: / / doi.org / 10.1124 / dmd.121.000463 (hereinafter “Ball 2022”), the entire contents of each of which are incorporated herein by reference for all purposes.5.4.4 Linkers

[0414] As described herein, the heterologous moiety of conjugates and fusion proteins and polypeptides described herein can be directly operably connected or indirectly operably connected to the CD161 binding protein (e.g., described herein).

[0415] In some embodiments, the heterologous moiety of conjugates and fusion proteins and polypeptides described herein is directly operably connected to the CD161 binding protein (e.g.,Attorney Docket No. 63340.32WO01described herein). In some embodiments, the heterologous moiety of conjugates and fusion proteins and polypeptides described herein is indirectly operably connected to the CD 161 binding protein (e.g., described herein). In some embodiments, the heterologous moiety of conjugates and fusion proteins and polypeptides described herein is indirectly operably connected to the CD161 binding protein (e.g., described herein) through a linker.

[0416] Linkers can be peptide linkers or non-peptide linkers. In some embodiments, the linker is derived from a crosslinking reagent. In some embodiments, the linker is cleavable. In some embodiments, the linker is non-cleavable. In some embodiments, the linker is synthetic.5.4.4.1 Non-peptide Linkers

[0417] In some embodiments, the linker is a non-peptide linker. In some embodiments, the linker is a chemical linker. In some embodiments, the linker is synthetic. In some embodiments, the linker is derived from a crosslinking reagent. In some embodiments, the linker is cleavable. In some embodiments, the linker is non-cleavable.

[0418] A variety of non-peptide linkers are known in the art that can be utilized to conjugate a heterologous moiety to a CD 161 binding protein described herein. Suitable linkers have two reactive termini, one for conjugation to a CD 161 binding protein described herein and one for conjugation to the heterologous moiety.

[0419] The terminus of the linker that is conjugated to a protein CD 161 binding protein described herein can be a site that is capable of conjugation to the protein through a cysteine thiol or lysine amine group on the protein, and as such comprises a thiol reactive group (e.g., as in maleimide, acrylamide, vinyl sulfone, bromide and tosylate linkers) or an amine-reactive group. The cysteine thiol or lysine amine group can be naturally occurring or engineered (e.g., proteins comprising engineered cysteine or lysine amino acid residues).

[0420] In some embodiments, the linker comprises one or more of polyethylene glycol (PEG), maleimide, acrylamide, vinyl sulfone, bromide or tosylate. In some embodiments, the linker is derived from a crosslinking reagent such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl 4-(2-pyridyldithio) pentanoate (SPP), N-succinimidyl 4-(2-pyridyldithio)butanoate (SPDB), N-succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), N-succinimidyl iodoacetate (SIA), N-succinimidyl (4-iodoacetyl)aminobenzoate (SIAB), maleimide PEG NHS, N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (sulfo-SMCC) or 2,5-Attorney Docket No. 63340.32WO01dioxopyrrolidin- 1 -yl 17-(2,5-dioxo-2,5-dihydro-l H-pyrrol-l-yl)-5,8,l l,14-tetraoxo-4,7,10,13-tetraazaheptadecan- 1 -oate (CX 1 - 1 ).

[0421] Exemplary linkers are for example described in e.g., Zheng Su et al., Antibody-drug conjugates: Recent advances in linker chemistry, Acta Pharmaceutica Sinica B, Volume 11, Issue 12, 3889-3907 (2021); and Sheyi, R.; de la Torre, B.G.; Albericio, F. Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate. Pharmaceutics 2022, 14, 396. https: / / doi.org / 10.3390 / pharmaceuticsl4020396, the entire contents of each of which is incorporated herein by reference for all purposes.5.4.4.2 Peptide Linkers

[0422] In some embodiments, the heterologous moiety e.g., heterologous protein) is directly operably connected to the CD161 binding protein e.g., described herein) via a peptide bond. In some embodiments, the heterologous moiety is indirectly operably connected to the CD 161 binding protein e.g., described herein) via a peptide linker.

[0423] In some embodiments, the linker is a peptide linker. In some embodiments, the peptide linker is one or any combination of a cleavable linker, a non-cleavable linker, a flexible linker, a rigid linker, a helical linker, and / or a non-helical linker.

[0424] In some embodiments, the peptide linker comprises from or from about 2-30, 5-30, 10-30, 15-30, 20-30, 25-30. 2-25, 5-25, 10-25, 15-25. 20-25. 2-20. 5-20, 10-20. 15-20. 2-15. 5-15, 10-15, 2-10, or 5-10 amino acid residues. In some embodiments, the peptide linker comprises at least about 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, or 30 amino acid residues. In some embodiments, the linker comprises or consists of about 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, or 30 amino acid residues. In some embodiments, the linker comprises or consists of no more than about 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, or 30 amino acid residues.

[0425] In some embodiments, the amino acid sequence of the peptide linker comprises or consists of glycine, serine, or both glycine and serine amino acid residues. In some embodiments, the amino acid sequence of the peptide linker comprises or consists of glycine, serine, and proline amino acid residues.Attorney Docket No. 63340.32WO01

[0426] The amino acid sequence of exemplary peptide linkers, which can be incorporated in one or more of the embodiments described herein (e.g., fusion proteins and polypeptides, conjugates), is set provided in Table 7.Table 7. The Amino Acid Sequence of Exemplary Peptide Linkers

[0427] In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of the linkers set forth in Table 7. In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of the linkers set forth in Table 7, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of the linkers set forth in Table 7, comprising 1, 2, or 3 amino acid variations (e.g., substitutions, deletions, additions). In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of the linkers set forth in Table 7, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid substitutions. In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the aminoAttorney Docket No. 63340.32WO01acid sequence of any one of the linkers set forth in Table 7, comprising 1, 2, or 3 amino acid substitutions.

[0428] In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39-58. In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39-58, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations {e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39-58, comprising 1, 2, or 3 amino acid variations {e.g., substitutions, deletions, additions). In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39-58, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%). amino acid substitutions. In some embodiments, the amino acid sequence of the peptide linker comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39-58, comprising 1, 2, or 3 amino acid substitutions.5.4.5 Orientation

[0429] The heterologous moiety {e.g., heterologous polypeptide) and the CD161 binding protein e.g., described herein) of a conjugate or fusion {e.g., described herein) can be arranged in any configuration or order as long as the CD161 binding protein {e.g., described herein) maintains the ability to mediate its function and the heterologous moiety {e.g., heterologous polypeptide) can mediate its function.

[0430] The heterologous moiety can be attached {e.g., conjugated or fused) to the N-terminus, C-terminus, or at an internal site {i.e., between the N- and C-terminus) of the CD161 binding protein {e.g., one or more polypeptide of a CD161 binding protein).

[0431] In some embodiments, the heterologous moiety is attached {e.g., conjugated or fused) to the N-terminus of the CD161 binding protein {e.g., one or more polypeptide of the CD161 binding protein) or polypeptide. In some embodiments, the heterologous moiety is attached {e.g., conjugated or fused) to the C-terminus of the CD161 binding protein {e.g., one or more polypeptide of the CD161 binding protein). In some embodiments, the heterologous moiety is attached {e.g.,Attorney Docket No. 63340.32WO01conjugated or fused) at an internal site (z.e., between the N- and C-terminus) of the CD161 binding protein (e.g., one or more polypeptide of the CD 161 binding protein).5.5 Multimeric CD161 Binding Proteins

[0432] In some embodiments, the CD161 binding proteins (e.g., described herein) (or conjugates or fusions comprising the same) are multimeric (e.g., dimeric, trimeric, tetrameric, etc.) proteins comprising at least two, three, or four polypeptides.

[0433] In some embodiments, the CD 161 binding protein comprises at least two, three, or four polypeptides. In some embodiments, the CD161 binding protein is dimeric (i.e., comprises two polypeptides). In some embodiments, the CD161 binding protein is trimeric (i.e., comprises three polypeptides). In some embodiments, the CD161 binding protein is tetrameric (i.e., comprises four polypeptides).

[0434] In some embodiments, two of the polypeptides associate via covalent or non-covalent interactions. In some embodiments, two of the polypeptides associate via at least one covalent interaction. In some embodiments, two of the polypeptides associate via one or more disulfide bond. In some embodiments, two of the polypeptides associate via 1, 2, 3, 4, or more disulfide bonds.

[0435] In some embodiments, the CD 161 binding protein comprises a full-length antibody comprising (i) a first Ig light chain comprising from N- to C-terminus a light chain variable region (VL) region and a light chain constant region (CL) region; (ii) a first Ig heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N- to C-terminus a VH region, a CHI region, a hinge region, a CH2 region, and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chain associate to form a first antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the amino acid sequence of the first heavy chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%. 92%, 93%, 94%. 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the first light chain is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%. 93%, 94%, 95%, 96%,Attorney Docket No. 63340.32WO0197%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain. In some embodiments, the amino acid sequence of the VH region of the first heavy chain is 100% identical to the amino acid sequence of the VH region of the second heavy chain; and the amino acid sequence of the first heavy chain outside of the VH region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second heavy chain outside of the VH region of the second heavy chain. In some embodiments, the amino acid sequence of the VL region of the first light chain is 100% identical to the amino acid sequence of the VL region of the second light chain; and the amino acid sequence of the first light chain outside of the VL region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the second light chain outside of the VL region of the second light chain.5.5.1 Exemplary Properties of CD161 Binding Proteins5.5.1.1 Affinity of CD161 Binding Proteins for CD161

[0436] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein)) exhibits a dissociation constant (Kd) for CD161 (e.g., hCD161) binding of < 10 pM, < 100 pM , < 10 pM , < 1 pM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM, and / or > 0.01 pM, 0.1 pM, or 1 pM (e.g. , 10-5M or less, 10-6 M or less, 10-8 M or less, e.g., from 1 pM to 10 pM, e.g., from 0.1 pM to 10 pM, e.g., from 10-6 M to 10-9 M, e.g., from 10-8 M to 10-13 M, e.g., from 10-9 M to 10-13 M) (e.g., as measured by SPR).

[0437] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of < 1 nM (e.g., as measured by SPR). In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of less than 1 nM (e.g., as measured by SPR). In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of from about 0.5nM - 1 nM (e.g., as measured by SPR).

[0438] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) exhibits a Kd for CD161 (e.g., hCD161) binding of < 1 nM. In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of less than 1 nM. In some embodiments, theAttorney Docket No. 63340.32WO01CD161 binding protein (e.g., an anti-CD16l antibody) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of from about 0.5nM - 1 nM.

[0439] Binding affinity can be measured by standard assays known in the art. For example, binding affinity can be measured by surface plasmon resonance (SPR) e.g., BIAcore®-based assay), a common method known in the art (see, e.g., Wilson. Science 295:2103, 2002; Wolff et al., Cancer Res. 55:2560. 1993; and U.S. Patent Nos. 5,283,173. 5,468.614, the full contents of each of which are incorporated by reference herein for all purposes). SPR measures changes in the concentration of molecules at a sensor surface as molecules bind to or dissociate from the surface. The change in the SPR signal is directly proportional to the change in mass concentration close to the surface, thereby allowing measurement of binding kinetics between two molecules (e.g., proteins). The dissociation constant for the complex can be determined by monitoring changes in the refractive index with respect to time as buffer is passed over the chip. In some embodiments, the affinity of a CD161 binding protein for CD161 (e.g., KD) is measured by SPR.

[0440] Other suitable assays for measuring the binding affinity include, for example, immunoassays such as enzyme linked immunosorbent assays (ELISA) and radioimmunoassays (RIA), or determination of binding by monitoring the change in the spectroscopic or optical properties of the proteins through fluorescence, UV absorption, circular dichroism, or nuclear magnetic resonance (NMR). Other exemplary assays include, but are not limited to, Western blot, analytical ultracentrifugation, spectroscopy, flow cytometry, sequencing and other methods for detection of binding of proteins.5.5.1.2 Inhibition of CD161 Binding to CLEC2D

[0441] In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) inhibits (blocks) the binding of CD161 to CLEC2D. In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) substantially inhibits (blocks) the binding of CD161 to CLEC2D. In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) inhibits (blocks) detectable the binding of CD161 to CLEC2D. In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) does not inhibit (block) the binding of CD161 to CLEC2D. In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) does not substantially inhibit (block) the binding of CD 161 to CLEC2D. In some embodiments, the CD161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein)Attorney Docket No. 63340.32WO01does not inhibit (block) detectable the binding of CD161 to CLEC2D.

[0442] In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of CD 161 to CLEC2D. In some embodiments, the CD161 binding protein e.g., hCD161 binding protein) {e.g., described herein)) partially inhibits binding of CD161 to CLEC2D. In some embodiments, the CD 161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) substantially inhibits binding of CD161 to CLEC2D. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) fully inhibits binding of CD161 to CLEC2D.

[0443] In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of hCD161 to hCLEC2D. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) partially inhibits binding of hCD161 to hCLEC2D. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) substantially inhibits binding of hCD161 to hCLEC2D. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) fully inhibits binding of hCD161 to hCLEC2D.

[0444] In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of < 10 pM, < 100 pM , < 10 pM , < 1 pM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM. < 0.01 nM, or < 0.001 nM, and / or > 0.01 pM, 0.1 pM, or 1 pM (e.g. , 10-5M or less. 10-6 M or less, 10-8 M or less, e.g., from 1 pM to 10 pM, e.g., from 0.1 pM to 10 pM, e.g., from 10-6 M to 10-9 M, e.g., from 10-8 M to 10-13 M, e.g., from 10-9 M to 10-13 M) {e.g., as measured by SPR).

[0445] In some embodiments, the {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of < 10 nM. In some embodiments, the CD 161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of less than 5 nM. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) inhibits {e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of less than 2 nM. In some embodiments, the CD161 binding protein {e.g., hCD161 binding protein) {e.g., described herein)) exhibits a Kd for CD161 {e.g., hCD161) binding of from about 0.5nM - 2 nM.Attorney Docket No. 63340.32WO01

[0446] In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CD 161 to CLEC2D and exhibits an IC50 of < 10 nM. In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of less than 5 nM (e.g., as measured by SPR). In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CD161 to CLEC2D and exhibits an IC50 of less than 2 nM. In some embodiments, the CD161 binding protein (e.g., hCD161 binding protein) (e.g., described herein)) exhibits a Kd for CD161 (e.g., hCD161) binding of from about 0.5nM - 2 nM.

[0447] The inhibition (blocking) (or lack thereof) of CD 161 binding to CLEC2D can be determined by standard methods known in the art. For example, an enzyme linked immunosorbent assays (ELISA). Standard binding assays to measure binding of CD 161 to CLEC2D are known in the art and described herein. See, e.g., Wade M, Mendez J, Coussens NP, et al. Inhibition of Protein-Protein Interactions: Cell-Based Assays. 2017 Nov 20. In: Markossian S, Grossman A, Brimacombe K, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Available from: https: / / www.ncbi.nlm.nih.gov / books / NBK464632 / ; Arkin MR, Glicksman MA, Fu H, et al. Inhibition of Protein-Protein Interactions: Non-Cellular Assay Formats. 2012 Mar 18 [Updated 2012 Oct 1]. In: Markossian S, Grossman A, Brimacombe K, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Available from: https: / / www.ncbi.nlm.nih.gov / books / NBK92000 / ; the entire contents of each of which are incorporated herein by reference for all purposes. For example, non-cell-based assays include, but are not limited to. ELISA. For further example, cellbased assays include, but are not limited to, energy transfer (Forster resonance energy transfer and bioluminescence resonance energy transfer) and protein complementation (fluorescence or enzymatic, e.g., luciferase).5.5.1.3 Depletion of Populations of CD161 Expressing Cells and Cytokines

[0448] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) is capable of mediating depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells. CD8+ T cells, Th2 cells, peTh2 cells, TH2AAttorney Docket No. 63340.32WO01cells, ILC2 cells), NK cells, B cells, MAIT cells (Mucosal Associated Innate T cells), gamma delta T cells (gd T cells), Tregs (Regulatory T cells), and / or ILCs (innate lymphoid cells))) (e.g., upon binding to CD161 expressed on the surface of the population of cells). In some embodiments, the population of CD 161 expressing cells comprises one or more subpopulations of immune cells. In some embodiments, the population of CD161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells). In some embodiments, the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells). , In some embodiments, the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells. In some embodiments, the population of CD161 expressing cells comprises TH2A cells.

[0449] In some embodiments, the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, monocytes, B cells, MAIT cells (Mucosal Associated Innate T cells), gamma delta T cells (gd T cells), Tregs (Regulatory T cells), and / or ILCs (innate lymphoid cells))) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 and IL-9). In some embodiments, the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the one or more proinflammatory cytokine is IL-4, IL-13, IL-5 or IL-9.

[0450] In some embodiments, the population of CD 161 expressing cells comprises CD8+ T cells and mediates a reduction in the level of one or more of IFNy, TNFa, IL- 17, IL-22, IL-21, or GM-CSF. In some embodiments, the population of CD161 expressing cells comprises TH2A cells and mediates a reduction in the level of one or more of IL-4, IL-13, IL-5 and IL-9.

[0451] In some embodiments, the CD161 binding protein (e.g., the anti-CD161 antibody) is capable of mediating a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9) (e.g., produced by a population of cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, monocytes, B cells, MAIT cells (Mucosal Associated Innate T cells), gamma delta T cells (gd T cells), Tregs (Regulatory T cells), and / or ILCs (inna...

Claims

Attorney Docket No. 63340.32WO01CLAIMSWhat Is Claimed Is:

1. A CD 161 binding protein comprising a binding domain that specifically binds CD161 (e.g., hCD161), wherein the binding domain comprises:(a) a VH region comprising (i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 3 or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid modifications; (ii) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 4 or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid modifications; and (iii) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 5 or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid modifications; and(b) a VL region comprising (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 6 or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid modifications; (ii) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 7 or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid modifications; and (iii) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 8 or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid modifications.

2. A CD161 binding protein comprising a binding domain that specifically binds CD161 (e.g., hCD161), wherein the binding domain comprises:(a) a VH region comprising (i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 3; (ii) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 4; and (iii) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 5; and (b) a VL region comprising (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 6; (ii) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 7; and (iii) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 8.

3. A CD161 binding protein comprising a binding domain that specifically binds CD161 (e.g., hCD161), wherein the binding domain comprises:Attorney Docket No. 63340.32WO01(a) a VH region comprising an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and(b) a VL region comprising an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

4. The CD 161 binding protein of claim 1 or 2, wherein(a) the amino acid sequence of the VH region is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and(b) the amino acid sequence of the VL region is at least 85%, 86%, 87%, 88%. 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

5. The CD 161 binding protein of any one of the preceding claims, wherein(a) the amino acid sequence of the VH region is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and(b) the amino acid sequence of the VL region is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

6. The CD161 binding protein of any one of the preceding claims, wherein(a) the amino acid sequence of the VH region is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and(b) the amino acid sequence of the VL region is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10.

7. The CD161 binding protein of any one of the preceding claims, wherein(a) the amino acid sequence of the VH region is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 9; and(b) the amino acid sequence of the VL region is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 10.

8. The CD161 binding protein of any one of the preceding claims, whereinAttorney Docket No. 63340.32WO01(a) the amino acid sequence of the VH region comprises the amino acid sequence set forth in SEQ ID NO: 9; and(b) the amino acid sequence of the VL region comprises the amino acid sequence set forth in SEQ ID NO: 10.

9. The CD161 binding protein of any one of the preceding claims, comprising a heavy chain (HC) and a light chain (LC).

10. The CD 161 binding protein of claim 9, wherein the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%. 89%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 67; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 68.

11. The CD 161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein (or the binding domain) comprises an antibody.

12. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein (or the binding domain) comprises an IgG {e.g., IgGi, IgGztc.y., IgGza or IgGzb), IgGs, IgG4), IgE, IgM, IgD, or IgA {e.g., IgAt or IgA2) antibody.

13. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein (or the binding domain) comprises one or more of a monoclonal antibody, monospecific antibody, multispecific antibody, human antibody, humanized antibody, chimeric antibody, and / or murine antibody, or a functional fragment or functional variant of any of the foregoing.

14. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein (or the binding domain) comprises one or more of a full-length antibody, scFv, Fab, F(ab')2, Fab'. Fv. single domain antibody {e.g., a VHH), scFv-Fc, Fab-Fc, and / or single domain antibody-Fc {e.g., VHH-Fc).

15. The CD 161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein {e.g., an anti-CD161 antibody) comprises a hlg Fc region.

16. The CD161 binding protein of any one of the preceding claims, wherein the hlg Fc region is capable of mediating one or more hlg Fc effector function.Attorney Docket No. 63340.32WO0117. The CD161 binding protein of any one of the preceding claims, wherein the hlg Fc region is capable of mediating one or more of ADCC, ADCP, or CDC.

18. The CD161 binding protein of any one of the preceding claims, wherein the hlg Fc region is capable of mediating ADCC.

19. The CD 161 binding protein of any one of the preceding claims, wherein the hlg Fc region comprises one or more amino acid variation or alteration (e.g., described herein (e.g., Table 4 herein)) in glycosylation that enhances one or more hlg Fc effector function (e.g., compared to a reference hlg Fc region or a reference CD 161 binding protein comprising the same).

20. The CD161 binding protein of any one of the preceding claims, wherein the hlg Fc region comprises one or more amino acid variation or alteration in glycosylation that enhances ADCC and / or ADCP (e.g., compared to a reference hlg Fc region or a reference CD161 binding protein comprising the same).

21. The CD 161 binding protein of any one of the preceding claims, wherein the hlg Fc region is afucosylated.

22. The CD161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein (e.g., an anti-CD161 antibody) (e.g., described herein) is capable of inhibiting (blocking) the binding of CD 161 to CLEC2D.

23. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein (e.g., the anti-CD161 antibody) is capable of mediating depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) (e.g., upon binding to CD161 expressed on the surface of the population of cells).

24. The CD 161 binding protein of any one of the preceding claims, wherein the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, GM-CSF, IL-4, IL- 13, IL-5 or IL-9).Attorney Docket No. 63340.32WO0125. The CD161 binding protein of any one of the preceding claims, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9.

26. The CD161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein {e.g., the anti-CD161 antibody) is capable of mediating a reduction in the level of one or more proinflammatory cytokine {e.g., IFNy, TNFa, IL- 17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9) {e.g., produced by a population of cells {e.g., immune cells {e.g., T cells {e.g., activated T cells) {e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs))).

27. The CD161 binding protein of any one of the preceding claims, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13. IL-5 or IL-9.

28. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein {e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector functions.

29. The CD 161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein {e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of antibody dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), or complement dependent cytotoxicity (CDC).

30. The CD161 binding protein of any one of the preceding claims, further comprising a heterologous moiety {e.g., a heterologous protein).

31. The CD161 binding protein of any one of the preceding claims, further comprising 2, 3, 4, or 5 or more heterologous moieties.

32. The CD 161 binding protein of any one of claims 30-31, wherein the heterologous moiety is attached to the N-terminus. C-terminus. and / or internally between the N- and C -terminus of the CD 161 binding protein.

33. The CD161 binding protein of any one of claims 30-32, wherein the heterologous moiety {e.g., heterologous polypeptide) is directly attached to the CD 161 binding protein.

34. The CD161 binding protein of any one of claims 30-33, wherein the heterologous moiety {e.g., heterologous polypeptide) is indirectly attached to the CD 161 binding protein.Attorney Docket No. 63340.32WO0135. The CD161 binding protein of any one of claims 30-34, wherein the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD 161 binding protein via a linker.

36. The CD161 binding protein of any one of claims 30-35, wherein the heterologous moiety is a peptide, protein (e.g., antibody), nucleic acid molecule (e.g., DNA, RNA), carbohydrate, lipid, polymer, or small molecule.

37. The CD161 binding protein of any one of claims 30-36, wherein the heterologous moiety is a half-life extension moiety (e.g., half-life extension protein (e.g., Fc region)).

38. The CD161 binding protein of any one of claims 30-37, wherein the heterologous moiety is a detectable tag and / or a reporter gene.

39. The CD161 binding protein of any one of claim 30-38, wherein the heterologous moiety is a therapeutic agent.

40. The CD161 binding protein of any one of the preceding claims, wherein the CD161 binding protein is isolated.

41. The CD 161 binding protein of any one of the preceding claims, wherein the CD 161 binding protein is recombinant.

42. A conjugate comprising the CD161 binding protein of any one of claims 1-41 and a heterologous moiety.

43. The conjugate of any one of claims 42, further comprising 2, 3, 4, or 5 or more heterologous moieties.

44. The conjugate of any one of claims 42-43, wherein the heterologous moiety is attached to the N-terminus, C-terminus, and / or internally between the N- and C-terminus of the CD161 binding protein.

45. The conjugate of any one of claims 42-44, wherein the heterologous moiety (e.g., heterologous polypeptide) is directly attached to the CD 161 binding protein.

46. The conjugate of any one of claims 42-45, wherein the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD 161 binding protein.

47. The conjugate of any one of claims 42-46, wherein the heterologous moiety (e.g., heterologous polypeptide) is indirectly attached to the CD 161 binding protein via a linker.

48. The conjugate of any one of claims 42-47, wherein the heterologous moiety is a peptide, protein, carbohydrate, lipid, polymer, or small molecule.Attorney Docket No. 63340.32WO0149. The conjugate of any one of claims 42-48, wherein the heterologous moiety is a therapeutic agent.

50. The conjugate of any one of claims 42-49, wherein the therapeutic agent is a small molecule.

51. A fusion protein comprising the CD161 binding protein of any one of claims 1-41 and a heterologous protein.

52. The fusion protein of claim 51, wherein the heterologous protein comprises a cytokine (or functional fragment or functional variant thereof), a chemokine (or a functional fragment or functional variant thereof), or an antibody (or a functional fragment or functional variant thereof).

53. The fusion protein of claim 51-52, wherein the heterologous protein is fused to the N-terminus, C-terminus, and / or internally between the N- and C-terminus of the CD161 binding protein.

54. The fusion protein of claim 51-53, wherein the heterologous protein is fused directly to the CD 161 binding protein.

55. The fusion protein of claim 51-54, wherein the heterologous protein is fused indirectly to the CD 161 binding protein.

56. The fusion protein of claim 51-55, wherein the heterologous protein is fused indirectly to the CD 161 binding protein via a peptide linker.

57. A nucleic acid molecule encoding the CD161 binding protein of any one of claims 1-41 or the fusion protein of any one of claims 51-56.

58. The nucleic acid molecule of claim 57, wherein the nucleic acid molecule is a DNA or RNA (e.g., mRNA) molecule.

59. The nucleic acid molecule of any one of claims 57-58, wherein the nucleic acid molecule is codon optimized.

60. The nucleic acid molecule of any one of claims 57-59, further comprising one or more transcription or translation regulatory elements (e.g., promoter, enhancer (e.g., cell or tissue specific transcription regulatory elements).

61. A vector comprising the nucleic acid molecule of any one of claims 57-60.

62. The vector of claim 61, wherein the vector is a viral vector or a non- viral vector (e.g., plasmid, minicircle).Attorney Docket No. 63340.32WO0163. The vector of any one of claims 61-62, wherein the vector is a viral vector (e.g., an adeno associated viral (AAV) vector, a lentiviral vector, an adenoviral vector).

64. A carrier comprising the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell (or population of cells) of claim 66, or the pharmaceutical composition of claim 67.

65. The carrier of claim 64, wherein the carrier is a nanoparticle, polymer, virus (e.g., a recombinant virus), virus like particle, virosome, fusosome, vesicle, or lipid-based carrier (e.g., a lipid nanoparticle (LNP), liposome, lipoplex, nanoliposome, an exosome, or a micelle).

66. A cell or population of cells comprising the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56. the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67.

67. A pharmaceutical composition comprising the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims OIOS, the cell or population of cells of claim 66, or the earner of any one of claims 64-65, and a pharmaceutically acceptable excipient.

68. A kit comprising the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the earner of any one of claims 64-65, or the pharmaceutical composition of claim 67; and optionally instructions for using any one or more of the foregoing.

69. A method of manufacturing the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56. the conjugate of any one of claims 42-50. the method comprising:introducing into a cell the nucleic acid molecule of any one of claim 57-60, the vector of any one of claims 61-63, or the carrier of any one of claims 64-65;culturing the cell under conditions that allow for expression of the CD 161 binding protein, the conjugate, or the fusion protein; andAttorney Docket No. 63340.32WO01optionally recovering the expressed the CD161 binding protein, the conjugate, or the fusion protein from the culture; andoptionally purifying the expressed the CD 161 binding protein, the conjugate, or the fusion protein from the culture.

70. A method of manufacturing an afucosylated version of the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56. the conjugate of any one of claims 42-50, the method comprising:introducing into a cell that comprises a functional disruption in the fucosyltransferase 8 (FUT8) gene the nucleic acid molecule of any one of claim 57-60, the vector of any one of claims 61-63, or the carrier of any one of claims 64-65;culturing the cell under conditions that allow for expression of the afucosylated CD 161 binding protein, the conjugate, or the fusion protein; andoptionally recovering the expressed the CD161 binding protein, the conjugate, or the fusion protein from the culture; andoptionally purifying the expressed the CD 161 binding protein, the conjugate, or the fusion protein from the culture.

71. A method of delivering a CD 161 binding protein described herein, a conjugate described herein, a fusion protein described herein, a nucleic acid molecule described herein, a vector described herein, a carrier described herein, a cell (or population of cells) described herein, or a pharmaceutical composition described herein to a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56. the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby deliver the CD161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or pharmaceutical composition to the subject.

72. A method of treating a proinflammatory disease (e.g., an autoimmune disease) in a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of anyAttorney Docket No. 63340.32WO01one of claims 61-63, the cell or population of cells of claim 66, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby treat the proinflammatory disease (e.g., the autoimmune disease) in the subject.

73. The method of claim 72, wherein the proinflammatory disease (e.g., the autoimmune disease) is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, an idiopathic inflammatory myopathies (e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

74. The method of claim 72, wherein the proinflammatory disease is an allergic disease (e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder (e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgE mediated disease (e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

75. The method of any one of claims 72-74, wherein the proinflammatory disease is an autoimmune disease.

76. The method of claim 75, wherein the autoimmune disease is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, an idiopathic inflammatory myopathies (e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

77. The method of claim 75, wherein the autoimmune disease is an allergic disease (e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder (e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgEAttorney Docket No. 63340.32WO01mediated disease (e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

78. The method of any one of claims 71-77, wherein the proinflammatory disease (e.g., an autoimmune disease) is treated, at least in part, through the depletion of a population of CD 161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells. MAIT cells, gd T cells. Tregs. and / or ILCs)) in the subject.

79. The method of any one of claims 71-78, wherein the CD161 binding protein (e.g., the anti-CD161 antibody) mediates depletion of a population of CD161 expressing cells (e.g., immune cells e.g., T cells e.g., activated T cells) e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) in the subject.

80. The method of any one of claims 71-79, wherein the depletion of the population of CD161 expressing cells (e.g., immune cells e.g., T cells (e.g., activated T cells) e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

81. The method of any one of claims 71-80, wherein the proinflammatory disease (e.g., an autoimmune disease) is treated, at least in part, through a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

82. The method of any one of claims 71-81, wherein the CD161 binding protein (e.g., the anti-CD161 antibody) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

83. The method of any one of claims 71-82, wherein the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9) is mediated by the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)).

84. The method of any one of claims 71-83, wherein the population of CD161 expressing cells comprises one or more subpopulations of immune cells.Attorney Docket No. 63340.32WO0185. The method of any one of claims 71 -84, wherein the population of CD161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).

86. The method of any one of claims 71-85, wherein the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells e.g., activated CD4+ T cells and / or CD8+ T cells).

87. The method of any one of claims 71-86. wherein the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

88. The method of any one of claims 71-87, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9.

89. A method of treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the earner of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby treat the autoimmune disease in the subject.

90. The method of claim 89, wherein the autoimmune disease is multiple sclerosis, psoriatic arthritis, hidradenitis suppurativa, polyarticular juvenile arthritis, Sjogren’s syndrome, psoriasis, alopecia areata, systemic sclerosis, one or more inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), ankylosing spondylitis, non-radiographic axial spondyloarthritis, systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, an idiopathic inflammatory myopathies (e.g., dermatomyositis, polymyositis, anti synthetase syndrome, inclusion-body myositis), sarcoidosis, enteropathic arthritis, or palmoplantar pustulosis.

91. The method of claim 89, wherein the autoimmune disease is an allergic disease (e.g., asthma, atopic dermatitis, allergic eosinophilic asthma, food allergy, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps (CRSwNP), an eosinophilic gastrointestinal disorder (e.g., eosinophilic esophagitis, hypereosinophilic syndrome, allergic rhinoconjunctivitis, an IgE mediated disease (e.g., bullous pemphigoid, lupus nephritis, pemphigus, autoimmune pancreatitis, and chronic spontaneous urticaria).

92. The method of any one of claims 89-91, wherein the autoimmune disease is treated, at least in part, through the depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells,Attorney Docket No. 63340.32WO01TH2A cells, ILC2 cells), NK cells, B cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) in the subject.

93. The method of any one of claims 89-92, wherein the CD161 binding protein (e.g., the anti-CD161 antibody) mediates depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells. MAIT cells, gd T cells. Tregs. and / or ILCs)) in the subject.

94. The method of any one of claims 89-93, wherein the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) mediates a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

95. The method of any one of claims 89-94, wherein the autoimmune disease is treated, at least in part, through a reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

96. The method of any one of claims 89-95, wherein the CD161 binding protein (e.g., the anti-CD161 antibody) mediates a reduction in the level of one or more prointlammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9).

97. The method of any one of claims 89-96, wherein the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9) is mediated by the depletion of the population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)).

98. The method of any one of claims 89-97, wherein the population of CD161 expressing cells comprises one or more subpopulations of immune cells.

99. The method of any one of claims 89-98, wherein the population of CD161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).

100. The method of any one of claims 89-99, wherein the population of CD161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells).Attorney Docket No. 63340.32WO01101. The method of any one of claims 89-100, wherein the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

102. The method of any one of claims 89-101, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9.

103. A method of depleting a population of cells (e.g., immune cells (e.g., T cells, NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) expressing CD161 e.g., on the surface) in a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby deplete the population of cells (e.g., immune cells (e.g., T cells, NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs)) expressing CD161 (e.g., on the surface) in the subject.

104. The method of claim 103, wherein the population of cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells, Tregs, and / or ILCs.

105. The method of claim 104, wherein the population of cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells, CD8+ T cells).

106. The method of claim 105, wherein the population of cells comprises CD4+ T cells and CD8+ T cells (e.g., activated CD4+ T cells and CD8+ T cells).

107. The method of any one of claims 103-106, wherein the population of cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

108. The method of any one of claims 103-107, wherein the population of cells expresses one or more proinflammatory cytokine.

109. The method of claim 108, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, GM-CSF, IL-4, IL- 13, IL-5 or IL-9.

110. The method of any one of claims 103-109, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) comprises a human Ig (hlg) Fc region.

111. The method of any one of claims 103-110, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector function.Attorney Docket No. 63340.32WO01112. The method of any one of claims 103- 111, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of antibody dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), or complement dependent cytotoxicity (CDC).

113. The method of any one of claims 103-112, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells, at least in part, through ADCC.

114. A method of reducing the level of one or more proinflammatory cytokines in a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby reduce the level of one or more proinflammatory cytokines in the subject.

115. The method of claim 114, wherein the one or more proinflammatory cytokine is IFNy, TNFa, IL- 17, IL-22, IL-21, GM-CSF, IL-4, IL- 13, IL-5 or IL-9.

116. The method of any one of claims 114-115, wherein the level of a plurality of proinflammatory cytokines is reduced (e.g., simultaneously).

117. The method of any one of claims 114- 116, wherein the level of at least 2, 3, 4, 5, or 6. or more proinflammatory cytokines is reduced (e.g., simultaneously).

118. The method of any one of claims 114-117, wherein the CD 161 binding protein (e.g., an anti-CD161 antibody) comprises a human Ig (hlg) Fc region.

119. The method of any one of claims 114-118, wherein the reduction in the level of one or more proinflammatory cytokine (e.g., IFNy, TNFa, IL-17, IL-22, IL-21, GM-CSF, IL-4, IL-13, IL-5 or IL-9) is mediated by the depletion of a population of CD161 expressing cells (e.g., immune cells (e.g., T cells (e.g., CD4+ T cells, CD8+ T cells, Th2 cells, peTh2 cells, TH2A cells, ILC2 cells), NK cells, MAIT cells, gd T cells. Tregs, and / or ILCs)).

120. The method of any one of claims 114-119, wherein the population of CD161 expressing cells comprises one or more subpopulations of immune cells.

121. The method of any one of claims 114-120, wherein the population of CD161 expressing cells comprises T cells (e.g., activated T cells) (e.g., CD4+ T cells and / or CD8+ T cells).Attorney Docket No. 63340.32WO01122. The method of any one of claims 114-121, wherein the population of CD 161 expressing cells comprises CD4+ T cells and / or CD8+ T cells (e.g., activated CD4+ T cells and / or CD8+ T cells).

123. The method of any one of claims 114-122, wherein the population of CD161 expressing cells comprises Th2 cells, peTh2 cells, TH2A cells, and / or ILC2 cells.

124. The method of any one of claims 119-123, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through one or more hlg Fc effector function.

125. The method of claim 124, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells through any one or more of ADCC, ADCP, or CDC.

126. The method of claim 125, wherein the CD161 binding protein (e.g., an anti-CD161 antibody) mediates depletion of the population of cells, at least in part, through ADCC.

127. A method of inhibiting (blocking) binding of CD161 to CLEC2D expressed on the surface of a cell in a subject in need thereof, the method comprising administering to the subject the CD161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66, the carrier of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby inhibit (blocking) binding of CD161 to CLEC2D expressed on the surface of a cell in the subject.

128. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject the CD 161 binding protein of any one of claims 1-41, the fusion protein of any one of claims 51-56, the conjugate of any one of claims 42-50, the nucleic acid molecule of any one of claims 57-60, the vector of any one of claims 61-63, the cell or population of cells of claim 66. the earner of any one of claims 64-65, or the pharmaceutical composition of claim 67, to thereby treat the cancer in the subject.

129. The method of claim 128, wherein the cancer is a leukemia or lymphoma.

130. The method of claim 128 or 129, wherein the cancer is a T cell leukemia, NK cell leukemia, T cell prolymphocytic leukemia (T-PLL), or large granular lymphocytic leukemia (LGLL).Attorney Docket No. 63340.32WO01131. The method of claim 128 or 129, wherein the cancer is a NK / T cell lymphoma (NKTCL), extranodal NK / T cell lymphoma (ENKL), mycosis fungoides (MF), Sezary syndrome, peripheral T cell lymphoma, angioimmunoblastic T cell lymphoma (AITL), and peripheral T cell lymphoma not otherwise specified (PTCL-NOS).

132. The method of any one of claims 71-131, wherein the CD 161 binding protein of any one of claims 1-41 or the fusion protein of any one of claims 51-56 is administered to the subject.

133. The method of any one of the preceding claims, wherein the CD161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or a pharmaceutical composition is administered in combination with one or more additional therapeutic agent.

134. The method of any one of claims 71-133, wherein the CD161 binding protein, conjugate, fusion protein, nucleic acid molecule, vector, carrier, cell (or population of cells), or a pharmaceutical composition is administered prior to, concomitant with, and / or after the administration of one or more additional therapeutic agent.

135. The method of any one of claims 133-134, wherein the one or more additional therapeutic agent is an anti-inflammatory agent.

136. The method of any one of claims 71-135, wherein the subject is human.

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