Anti-CD161 antibody and method of use thereof

Abstract

This disclosure provides antibodies that bind to CD161 and their antigen-binding moieties, bispecific and multispecific antibodies containing them, nucleic acids and vectors encoding them, and methods for using them to treat subjects in need. In some embodiments, the anti-CD161 antibodies and their antigen-binding moieties described herein block or reduce the interaction between CD161 and CLEC2D. The binding of CD161 to CLEC2D inhibits T cell activation.

Description

[Technical Field]

[0001] Cross-reference of related applications This PCT application claims priority to U.S. Provisional Patent Application No. 63 / 500,122, filed 4 May 2023, which is incorporated herein by reference in its entirety.

[0002] Reference to electronically submitted sequence listings The contents of the sequence listing were submitted electronically with this application (title: 5037_001PC01_Seqlisting_ST26; size: 304,752 bytes; and creation date: May 3, 2024), and are incorporated herein by reference in their entirety.

[0003] field This disclosure relates to an antibody that binds to CD161, its antigen-binding moiety, and a method for using it to treat a target that requires it. [Background technology]

[0004] background Human cancers involve numerous genetic and epigenetic alterations, leading to neoantigens potentially recognizable by the immune system (Sjoblom et al., Science 2006) 314(5797):268-274). The adaptive immune system, composed of T and B lymphocytes, possesses a broad ability to respond to diverse tumor antigens and excellent specificity, resulting in potent anti-cancer activity. The use of immunotherapy in cancer treatment is based on the premise that tumors evade the endogenous immune response by being recognized as self and non-self. Tumors can evade immune surveillance and acquire immune resistance using various mechanisms. Novel approaches in cancer immunotherapy aim to counter these resistance mechanisms and enable the endogenous immune system to eliminate the tumor. Recent successes of immunomodulatory agents, such as anti-PD-1 and anti-CTLA-4 therapies, in patients with refractory solid tumors have demonstrated the concept that immune system activation is effective as immunotherapy. Despite these advances in immunotherapy, not all tumors respond to approved immunomodulators (Filley et al. 2017). Furthermore, tumors form a suppressive microenvironment to evade and inhibit immune responses, leading to variability in the effectiveness of anticancer drugs based on patient-specific characteristics (Topalian et al. 2014). Immune resistance to current immunotherapies remains a significant challenge. Therefore, further therapies are needed that can induce immune responses targeting cancer and other malignancies in an immunosuppressive environment. Immunotherapy in combination with radiation therapy, chemotherapy, and / or surgery can provide effective treatment options for cancer. Novel drug-worthy targets and therapeutic combinations will play a crucial role in expanding the possibilities of immunotherapy in the treatment of various cancers. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Sjoblom et al.,Science 2006)314(5797):268-274 [Overview of the project]

[0006] Brief Overview Some aspects of this disclosure relate to an antibody or its antigen-binding moiety that specifically binds to CD161, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein VH comprises VH complementarity-determining region 1 (VH-CDR1), VH-CDR2, and VH-CDR3, and VL comprises VL-CDR1, VL-CDR2, and VL-CDR3, and VH-CDR3 comprises SEQ ID NO: 3, 1 This relates to an antibody or its antigen-binding moiety, comprising an amino acid sequence selected from the sequences described in 3, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323, and 333.

[0007] In some embodiments, VH-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, and 332. In some embodiments, VH-CDR1 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, and 331. In some embodiments, VL-CDR3 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, and 336. In some embodiments, VL-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, and 335. In some embodiments, VL-CDR1 comprises an amino acid sequence selected from the sequences described in Sequence IDs 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, and 334.

[0008] In some embodiments, the antibody or its antigen-binding portion is (i) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (ii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96; (iii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, and VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113 VL-CDR1 containing the amino acid sequence described in CDR3, SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116; (iv) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, and SEQ ID NO: 145 VL-CDR2 containing the amino acid sequence, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146; (v) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216;(vi) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226; (vii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, and the amino acid sequence described in SEQ ID NO: 272 VH-CDR2 containing the sequence, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276; (viii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, and SEQ ID NO: 284 VL-CDR1 containing the amino acid sequence described, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286; (ix) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and sequence VL-CDR3 containing the amino acid sequence described in number 296; (x) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306; or (xi) any combination of (i) to (x).

[0009] In some embodiments, VH includes an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. In some embodiments, VL includes an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

[0010] Some aspects of the present disclosure relate to an antibody or antigen-binding moiety that specifically binds to CD161, comprising a variable heavy chain (VH) domain and a variable light chain (VL) domain, wherein the VH is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or with respect to an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. VL relates to an antibody or its antigen-binding moiety, which contains an amino acid sequence having at least approximately 99% sequence identity, and VL contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

[0011] In some embodiments, VH comprises an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. In some embodiments, VL comprises an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

[0012] In some embodiments, (i) VH includes an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 7, and VL includes an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 8; (ii) VH includes an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 97 (iii) VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 98; (iii) VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 117, and VL includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 118;(iv) VH contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 147, and VL contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 148; (v) VH contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 217 (i) VL contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 218; (vi) VH contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 227, and VL contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 228;(vii) VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 277, and VL includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 278; (viii) VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 287 (ix)VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 288; (ix)VH includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 297, and VL includes an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 298;(x)VH contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 307, and VL contains an amino acid sequence having at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, or at least approximately 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 308; or (xi) any combination of (i) to (x).

[0013] In some embodiments, the antibody or its antigen-binding portion is (i) VH containing the amino acid sequence described in SEQ ID NO: 7 and VL containing the amino acid sequence described in SEQ ID NO: 8; (ii) VH containing the amino acid sequence described in SEQ ID NO: 97 and VL containing the amino acid sequence described in SEQ ID NO: 98; (iii) VH containing the amino acid sequence described in SEQ ID NO: 117 and VL containing the amino acid sequence described in SEQ ID NO: 118; (iv) VH containing the amino acid sequence described in SEQ ID NO: 147 and VL containing the amino acid sequence described in SEQ ID NO: 148; (v) VH containing the amino acid sequence described in SEQ ID NO: 217 and VL containing the amino acid sequence described in SEQ ID NO: 218; (vi) (vii) VH containing the amino acid sequence described in sequence number 227 and VL containing the amino acid sequence described in sequence number 228; (viii) VH containing the amino acid sequence described in sequence number 277 and VL containing the amino acid sequence described in sequence number 278; (ix) VH containing the amino acid sequence described in sequence number 297 and VL containing the amino acid sequence described in sequence number 298; (x) VH containing the amino acid sequence described in sequence number 307 and VL containing the amino acid sequence described in sequence number 308; or (xi) any combination of (i) to (x). Some aspects of this disclosure relate to antibodies or antigen-binding moieties that bind to the same epitope as the antibodies or antigen-binding moieties disclosed herein. Some aspects of this disclosure relate to antibodies or antigen-binding moieties that cross-compete with the antibodies or antigen-binding moieties disclosed herein for binding to CD161.

[0014] In some embodiments, the antibody or its antigen-binding moiety has a K content of less than approximately 1000 nM, less than approximately 500 nM, less than approximately 100 nM, less than approximately 50 nM, or less than approximately 10 nM. D It binds to CD161. In some embodiments, the antibody or its antigen-binding moiety is less than 50 nM of K D It binds to CD161. In some embodiments, the antibody or its antigen-binding moiety is less than 10 nM of K D Then combine it with CD161.

[0015] In some embodiments, the antibody or its antigen-binding moiety inhibits the interaction between CD161 and member D of the C-type lectin domain family 2 (CLEC2D).

[0016] In some embodiments, an antibody or its antigen-binding moiety can induce or enhance the production of one or more cytokines by immune cells. In some embodiments, the one or more cytokines include IL2, TNFα, IFNg, or any combination thereof.

[0017] In some embodiments, the antigen-binding portion of the antibody includes a VHH, vNAR, microbody, nanobody, scFv, or any combination thereof.

[0018] Some aspects of this disclosure relate to multispecific antibodies comprising the antibody or its antigen-binding moiety disclosed herein. Some aspects of this disclosure relate to bispecific antibodies comprising the antibody or its antigen-binding moiety disclosed herein.

[0019] Some aspects of this disclosure relate to nucleic acid molecules or sets of nucleic acid molecules encoding an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, or a bispecific antibody disclosed herein.

[0020] Some aspects of this disclosure relate to vectors or sets of vectors, including nucleic acid molecules or sets of nucleic acid molecules disclosed herein. In some aspects, the vectors are viral vectors.

[0021] Some aspects of this disclosure relate to host cells, including nucleic acid molecules or sets of nucleic acid molecules disclosed herein, or vectors or sets of vectors disclosed herein.

[0022] Some aspects of this disclosure relate to pharmaceutical compositions comprising an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, or a host cell disclosed herein, and a pharmaceutically acceptable carrier.

[0023] Some aspects of this disclosure relate to methods for treating a disease or disorder in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the disease or disorder includes cancer.

[0024] Some aspects of this disclosure relate to methods for inducing an immune response in a subject requiring such induction, comprising administering to the subject an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the subject is suffering from cancer.

[0025] Some aspects of this disclosure relate to methods for treating cancer in a subject in need of such treatment, comprising administering to the subject an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein.

[0026] In some aspects, cancer includes acoustic neuroma, acute lymphoblastic leukemia, acute myeloid leukemia, adenocarcinoma, and cancers of the urinary tract, as well as carcinomas, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, biliary tract cancer, bladder cancer, bone cancer, brain cancer, brainstem glioma, breast cancer, bronchogenic carcinoma, Burkitt lymphoma and marginal zone B-cell lymphoma, adrenal cancer, anal cancer, gastrointestinal cancer, endocrine cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical carcinoma, endometrial carcinoma, fallopian tube carcinoma, renal pelvis carcinoma, vaginal carcinoma, vulvar carcinoma, and central nervous system CNS cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, craniopharyngioma, cystadenocarcinoma, fetal cancer, endometrial cancer, endosarcoma, environmentally induced cancer (including those induced by asbestos), ependymoma, epidermal carcinoma, epithelial carcinoma, esophageal cancer, esophageal carcinoma, Ewing's tumor, ocular cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, germ cell tumor, glioblastoma (e.g., glioblastoma multiforme), glioma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatoma, Hodgkin's disease, carcinoma in situ, Kaposi's sarcoma, kidney cancer (e.g.) Renal cell carcinoma (RCC), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung carcinoma, intralymphatic sarcoma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyeloblastic myelomonocytic monocytic erythroleukemia, myxosarcoma, nasopharyngeal cancer, neoplasms of the central nervous system (CNS), neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer (NSCLC), non-small cell lung carcinoma, oligodendroglioma, oral cancer (e.g., lips, tongue, mouth and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary carcinoma, papillary carcinoma, pediatric sarcoma, pineal gland Tumors, pituitary adenoma, polycythemia vera lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate cancer), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, soft tissue sarcoma, sebaceous gland carcinoma, seminomas, nasal cavity / sinus natural killer cancers, skin cancer, small cell lung cancer (SCLC), pediatric solid tumors, spinal axial tumors, squamous cell carcinoma, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synoviomas, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-associated cancer or cancer of virus origin (e.g., human papillomavirus (HPV-associated or HPV-derived tumors)),Waldenström macroglobulinemia and Wilms' tumor; as well as any combination of the aforementioned cancers.

[0027] Some aspects of this disclosure relate to methods for treating infectious diseases in subjects in need thereof, comprising administering to such subjects an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the infectious disease is (i) an infection caused by influenza, herpes, giardia, malaria, leishmania, or any combination thereof; (ii) human immunodeficiency virus (HIV), hepatitis virus, herpesvirus, adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, (iii) Infections caused by molluscum contagiosum virus, poliovirus, rabies virus, JC virus, or arbovirus encephalitis virus, or any combination thereof; (iii) Infections caused by Chlamydia, Rickettsia, Mycobacteria, Staphylococcus, Streptococcus, Pneumococcus, Meningococcus, Conococcus, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacillus, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease, or any combination thereof; (iv) Candida, Cryptococcus Infections caused by neoformans, Aspergillus, Mucorales, Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, or Histoplasma capsulatum, or any combination thereof; (v) Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp.Infections caused by Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, or Nippostrongylus brasiliensis, or any combination thereof; or including any combination of (vi)(i)-(v).

[0028] Some aspects of this disclosure relate to methods for treating autoimmune diseases in subjects in need thereof, comprising administering to such subjects an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein.

[0029] Some aspects of this disclosure relate to methods for activating immune cells, which include contacting the immune cells with an antibody or antigen-binding moiety thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or set of nucleic acid molecules disclosed herein, a vector or set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. [Brief explanation of the drawing]

[0030] [Figure 1]The images show violin plots of scRNA-seq gene expression profiles for killer cell lectin-like receptor B1 protein (KLRB1 / CD161), PDCD1, LAG3, CTLA4, HAVCR2, and TIGIT in CD4 T cells, CD8 T cells, dendritic cells (DCs), macrophages, monocytes, naive B cells, NK cells, and regulatory T cells (Tregs) derived from specific tumor samples.

[0031] [Figure 2] The image shows violin plots of scRNA-seq gene expression for CD161, PDCD1, LAG3, CTLA4, HAVCR2, and TIGIT in naive-like CD8 T cells, early-active CD8 T cells, effector memory CD8 T cells, CD8 Tpex cells, and CD8 Tex cells in specific tumor samples.

[0032] [Figure 3] This graph shows cell-based binding of anti-CD161 positive hits / antibodies that cross-react with human and cyno CD161 proteins expressed on CHOK1 cells, as measured by flow cytometry.

[0033] [Figure 4] This graph shows the binding of anti-CD161 antibodies specific to human CD161 (hCD161) protein in an ELISA assay.

[0034] [Figure 5A] This is a FACS plot of cell-based binding using isotype controls. [Figure 5B] This is a cell-base bound FACS plot using an anti-CD161 positive antibody (01D17). [Figure 5C] This is a cell-base bound FACS plot using an anti-CD161 positive antibody (03K18). [Figure 5D] This graph shows the binding of various anti-CD161 antibodies to human CD161-expressing CHOK1 cells.

[0035] [Figure 6] This graph shows the binding of human CLEC2D multimers to human CD161-expressing CHOK1 cells. The anti-CD161 reference monoclonal antibody HP-3G10 blocks the binding of hCLEC2D multimers to hCD161-expressing CHOK1 cells.

[0036] [Figure 7] This bar graph shows the percentage of blockade of the hCD161:hCLEC2D interaction by anti-CD161 antibodies using a cell-based assay.

[0037] [Figure 8] This graph shows the concentration-dependent inhibition of the hCD161:hCLE2CD interaction by various anti-CD161 antibodies in an ELISA assay.

[0038] [Figure 9] This bar graph shows IL-2 cytokine release in healthy human PBMCs (Plantobozoic Microorganisms) stimulated with SEB (Staphylococcal Enterotoxin B) and then exposed to various anti-CD161 antibodies, as measured by ELISA.

[0039] [Figure 10] This bar graph shows IL-2 cytokine release in hCD161-expressing Jurkat cells that were activated by T cell engagers and Raji cells and then treated with various anti-CD161 antibodies, as measured by ELISA.

[0040] [Figure 11] This is a schematic diagram illustrating TCR activation affected by the CD161-CLEC2D interaction, using Jurkat-MART-1 TCR cells and MeWo cells as examples.

[0041] [Figure 12]This graph shows TCR activation (measured by IL-2 release (pg / mL)) in MART1 TCR-specific T cells contacted with hCLEC2D-GFP overexpressing MeWo cells (HLA; A*0201) in the presence of various concentrations of the anti-CD161 antibody described herein. The upper dashed line (1) represents the IL-2 release level when Jurkat-MART1 TCR CD161 cells are contacted with mock (GFP) MeWo cells (control, no impairment of TCR activity). The lower dashed line (2) represents the IL-2 release level when Jurkat-MART1 TCR CD161 cells are contacted with hCLEC2D-GFP overexpressing MeWo cells in the absence of anti-CD161 antibody (control, with TCR activity inhibition).

[0042] [Figure 13] This graph shows the dose-dependent binding of an anti-CD161 antibody to TALL-104 effector cells expressing CD161.

[0043] [Figure 14A] A schematic diagram of a cytotoxic assay is shown. [Figure 14B] Preliminary data shows that this cytotoxic assay can detect changes in cell lysis induced by TALL-104 in MeWo cells. [Figure 14C] Preliminary data shows that this cytotoxic assay can detect changes in cell lysis induced by TALL-104 in PC3 cells.

[0044] [Figure 15]This graph shows the percentage of cell lysis (%) by TALL-104 effector cells against human CLEC2D-overexpressing PC3 target cells in the presence of gradually increasing doses of anti-CD161 monoclonal antibody. The upper dashed line (1) represents the level (%) of TALL-104 cell lysis against MOCK-PC3 cells (these cells do not express human CLEC2D and therefore do not inhibit TALL-104 effector cells). The lower dashed line (2) represents the level (%) of TALL-104 cell lysis (%) against CLEC2D-overexpressing PC3 target cells in the absence of anti-CD161 antibody.

[0045] [Figure 16] A and B are dot plots showing the relative tumor-killing ability obtained from co-culture of primary NK cells and PC-3 CLEC2D-OE cells (CLEC2D overexpressing; A) or Raji cells (B) in the presence of anti-CD161 mAb. *p<0.05, ****p<0.00001, determined by one-way ANOVA with multiple comparisons.

[0046] [Figure 17] This graph shows the PC3-CLEC2D-OE tumor volume (mm3) in NSG mice after two intratumoral injections of NK cells and administration of vehicle or anti-CD161 antibody 12G06. n=4 per group. *p≦0.05, determined by two-way ANOVA.

[0047] [Figure 18] This graph shows the U87 tumor volume (mm3) in NSG mice after co-transplantation with human PBMCs and administration of vehicle or anti-CD161 antibody 12G06. n=4 per group. *p≦0.05, determined by two-way ANOVA. [Modes for carrying out the invention]

[0048] Detailed explanation This disclosure relates to antibodies that specifically bind to CD161, referred to herein as anti-CD161 antibodies, and their antigen-binding moieties. Some aspects of this disclosure relate to bispecific or multispecific antibodies, including anti-CD161 antibodies. Some aspects of this disclosure relate to methods for treating a disease or disorder in a subject in need thereof, comprising administering an antibody or its antigen-binding moiety described herein to the subject.

[0049] Prior to a detailed description of this disclosure, it should be understood that this disclosure is not limited to any specific composition or process step described, and that these are, of course, different. As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has separate components and features that can be readily separated from or combined with any of the features of several other embodiments without departing from the scope or spirit of this disclosure. Any of the enumerated methods may be carried out in the order of the enumerated events or in any other logically possible order.

[0050] The headings provided herein are not intended to limit the various aspects of this disclosure, which can be defined by referring to this entire specification. Furthermore, the terms used herein are intended solely to describe specific aspects and not to limit them.

[0051] I. Terminology To facilitate understanding of this explanation, we will first define some terms. Further definitions will be provided throughout the section on "Modes for Carrying Out the Invention."

[0052] It should be noted that the term "a" or "an" entity refers to one or more of those entities; for example, "a nucleotide sequence" is understood to represent one or more nucleotide sequences. Therefore, the terms "a" (or "an"), "one or more," and "at least one" may be used interchangeably in this specification.

[0053] Furthermore, as used herein, “and / or” shall be considered a specific disclosure of each of two designated features or components, whether one is accompanied by the other or not. Accordingly, the term “and / or” as used herein in expressions such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to include each of the following embodiments: 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).

[0054] Whenever an aspect is described herein together with the stylistic phrase “including,” it is understood that other similar aspects described in terms of “consisting of” and / or “essentially consisting of” are also provided.

[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in the field relating to this disclosure. For example, *Concise Dictionary of Biomedicine and Molecular Biology*, Juo, Pei-Show, 2nd ed., 2002, CRC Press; *The Dictionary of Cell and Molecular Biology*, 3rd ed., 1999, Academic Press; and *Oxford Dictionary of Biochemistry and Molecular Biology*, Revised, 2000, Oxford University Press provide many general dictionaries of the terms used herein.

[0056] Units, prefixes, and symbols are shown in their SI-approved forms. Numerical ranges include the number defining the range. Unless otherwise indicated, nucleotide sequences are written from left to right in the 5' to 3' direction. Amino acid sequences are written from left to right in the amino to carboxy direction. The headings provided herein are not intended to limit the various aspects of this disclosure that can be obtained by referring to the entire specification. Thus, the terms defined immediately below are more fully defined by referring to the entire specification.

[0057] The term "approximately" is used herein to mean roughly, about, approximately, or within a range. When the term "approximately" is used with a numerical range, it modifies that range by extending the boundary values ​​above and below the stated numerical value. Generally, the term "approximately" may modify a numerical value above or below the stated value by, for example, a variation of 10 percent above or below (higher or lower).

[0058] In some embodiments, the term “antibody” refers to a protein or its antigen-binding portion comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region (abbreviated herein as CH). In some antibodies, such as naturally occurring IgG antibodies, the heavy chain constant region consists of a hinge and three domains: CH1, CH2, and CH3. In some antibodies, such as naturally occurring IgG antibodies, each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (abbreviated herein as CL). The VH and VL regions can be further divided into a highly variable region called the complementarity-determining region (CDR) and a more conserved region called the framework region (FR) interspersed between them. Each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1q). The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified herein, the amino acids in the variable region are numbered using the Kabat numbering system, and the amino acids in the constant region are numbered using the EU system.

[0059] "IgG antibodies", such as human IgG1, IgG2, IgG3, and IgG4 antibodies, when used herein, in some embodiments, have the structure of naturally occurring IgG antibodies, i.e., they have the same number of heavy and light chains and disulfide bonds as naturally occurring IgG antibodies of the same subclass. For example, an anti-CD161 IgG1, IgG2, IgG3, or IgG4 antibody consists of two heavy chains (HC) and two light chains (LC), where the two HC and LC are linked by disulfide bonds of the same number and in the same positions as those present in naturally occurring IgG1, IgG2, IgG3, and IgG4 antibodies (except when the antibody is mutated and the disulfide bridges are modified).

[0060] Antibodies typically bind specifically to cognate antigens with high affinity, which is represented by a dissociation constant (K -5 ~10 -11 ) of 10 D M or less. A K -4 exceeding about 10 D M is generally considered to indicate non-specific binding. As used herein, an antibody that "binds specifically" to an antigen refers to an antibody that binds to the antigen and substantially identical antigens with high affinity, which means that the K D is 10 -7 M or less, 10 -8 M or less, 5×10 -9 M or less, or 10 -8 M to 10 -10 M or less, and does not bind to unrelated antigens with high affinity.

[0061] Immunoglobulins may be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG isotypes are classified into subclasses in certain species; in humans, they are classified into IgG1, IgG2, IgG3, and IgG4; and in mice, into IgG1, IgG2a, IgG2b, and IgG3. In some embodiments, the anti-CD161 antibodies described herein are of the IgG1 subtype. Immunoglobulins, such as IgG1, exist in several allotypes, differing from each other by at most a few amino acids. “Antibody” includes, for example, naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human and non-human antibodies, as well as fully synthetic antibodies.

[0062] The term “antigen-binding moiety” of an antibody, as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., human CD161). It has been shown that the antigen-binding function of an antibody can also be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term “antigen-binding moiety” of an antibody, e.g., an anti-CD161 antibody as described herein, include (i)V L , V H (ii) Fab fragments (fragments obtained from papain cleavage) or similar monovalent fragments consisting of LC and CH1 domains; (ii) F(ab')2 fragments (fragments obtained from pepsin cleavage) or similar divalent fragments containing two Fab fragments linked by disulfide bridges in a hinge region; (iii) V H and Fd fragment consisting of CH1 domain; (iv) V of a single arm of antibody L and V H Fv fragment consisting of domains; (v)V H Examples include (vi) a dAb fragment consisting of domains (Ward et al., (1989) Nature 341:544-546); (vi) an isolated complementarity-determining region (CDR); and (vii) a combination of two or more isolated CDRs that can be linked by a synthetic linker of any choice. Furthermore, there are two domains of the Fv fragment VL and V H Although encoded by separate genes, they can be constructed as a single protein chain by linking them together with a synthetic linker using recombination. L Region and V H The regions pair up to form a monovalent molecule (known as a single-stranded Fv (scFv)) (see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-stranded antibodies are also intended to be included in the term "antigen-binding moiety" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and these fragments are screened for utility in the same manner as intact antibodies. Antigen-binding moieties can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins.

[0063] A "bispecific antibody" or "bifunctional antibody" is an artificial hybrid antibody having two different heavy / light chain pairs and two different binding sites. A "multispecific antibody" or "multifunctional antibody" is an artificial hybrid antibody having three or more different heavy / light chain pairs and two different binding sites. Bispecific and multispecific antibodies can be produced by various methods, including hybridoma fusion or Fab' fragment linking. See, for example, Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0064] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies in that population are substantially similar and bind to the same epitope (for example, the antibody exhibits a single binding specificity and affinity). The exception is variants that may arise during the production of monoclonal antibodies, which are usually present, if any. The modifier “monoclonal” indicates a characteristic of the antibody that it is obtained from a substantially homogeneous antibody population and should not be interpreted as requiring the production of the antibody by any particular method. The term “human monoclonal antibody” refers to an antibody obtained from a substantially homogeneous antibody population exhibiting a single binding specificity and having a variable region derived from a human germline immunoglobulin sequence and an arbitrary constant region. In some embodiments, human monoclonal antibodies may be produced by hybridomas obtained by fusing B cells from transgenic non-human animals, such as transgenic mice having a genome containing human heavy chain and light chain transgenes, with immortalized cells.

[0065] The term “recombinant human antibody” as used herein includes all human antibodies prepared, expressed, produced or isolated by recombinant means, for example: (a) antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomes with respect to human immunoglobulin genes or hybridomas prepared therefrom; (b) antibodies isolated from host cells transformed to express antibodies, for example, transfectomas; (c) antibodies isolated from recombinant combinatorial human antibody libraries; and (d) antibodies prepared, expressed, produced or isolated by any other means involving the incorporation of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies include variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but also include, for example, subsequent rearrangements and mutations that occur during the maturation process of the antibody. As is known in the art (see, for example, Lonberg (2005) Nature Biotech. 23(9):1117-1125), the variable region contains antigen-binding domains encoded by various genes, and the rearrangement of these genes leads to the formation of antibodies specific to foreign antigens. In addition to rearrangement, the variable region can be further modified by multiple single amino acid changes (called somatic mutations or hypermutations) to increase the affinity of antibodies to foreign antigens. The constant region changes in further responses to the antigen (i.e., isotype switches). Therefore, nucleic acid molecules encoding light-chain and heavy-chain immunoglobulin polypeptides that have been rearranged and somatically mutated in response to an antigen cannot have sequence identity with the original nucleic acid molecule, but are substantially identical or similar (i.e., have at least 80% identity).

[0066] A “human” antibody (HuMAb) refers to an antibody having a variable region in which both the framework region and the CDR region are derived from a human germline immunoglobulin sequence. Furthermore, if the antibody contains a constant region, that constant region is also derived from a human germline immunoglobulin sequence. Anti-CD161 antibodies described herein may contain amino acid residues not encoded by a human germline immunoglobulin sequence (e.g., mutations introduced by random mutagenesis or site-directed mutagenesis in vitro or somatic mutagenesis in vivo). However, as used herein, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, is grafted onto a human framework sequence. The terms “human” antibody and “fully human” are used synonymously.

[0067] A "humanized antibody" refers to an antibody in which some, almost all, or all of the amino acids outside the CDR domain of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulin. In some embodiments of humanized antibodies, some, almost all, or all of the amino acids outside the CDR domain are replaced with amino acids from human immunoglobulin, but some, almost all, or all of the amino acids within one or more CDR regions remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are acceptable as long as they do not impede the antibody's ability to bind to a particular antigen. "Humanized" antibodies retain similar antigen specificity to the original antibody.

[0068] A "chimeric antibody" refers to an antibody in which the variable region originates from one species and the constant region originates from another species, such as an antibody in which the variable region originates from a mouse antibody and the constant region originates from a human antibody.

[0069] As used herein, “isotype” refers to an antibody class encoded by a heavy chain constant region gene (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE antibodies).

[0070] In this specification, the terms "antibody that recognizes an antigen" and "antibody that is specific to an antigen" are used interchangeably with the term "antibody that specifically binds to an antigen."

[0071] As used herein, "isolated antibody" is intended to refer to an antibody that is substantially free of other proteins and cellular material.

[0072] As used herein, “CD161” refers to the polypeptide encoded by the human KLRB1 gene, which is expressed in a subset of NK cells and peripheral blood T cells. CD161 may also be referred to as “Killer Cell Lectin-like Receptor Subfamily B Member 1,” “KLRB1,” “Type C Lectin Domain Family 5 Member B,” “CLEC5B,” “Type C Lectin Domain Family 5 Member B,” and “NKRP1a.” CD161 is thought to inhibit NK cell-mediated cytotoxicity and interferon-gamma secretion from target cells by binding to CLEC2D / LLT1. Activated CD161 leads to specific acidic sphingomyelinase (aSMase) stimulation, resulting in a significant increase in intracellular ceramide. CD161 acts as a lectin that binds to the terminal glycan Gal-alpha(1,3)Gal epitope and N-acetyllactosamine epitope. Standard human CD161 sequences are provided in Table 1 (UniProt Q12918). [Table 1]

[0073] As used herein, an antibody that inhibits "CD161 activity" is intended to refer to an antibody that inhibits or reduces one or more activities of CD161. In some embodiments, the anti-CD161 antibodies disclosed herein inhibit or reduce the interaction between human CD161 and CLEC2D. In some embodiments, the anti-CD161 antibodies disclosed herein increase NK cell-mediated cytotoxicity, i.e., eliminate CD161-mediated inhibition. In some embodiments, the anti-CD161 antibodies disclosed herein increase interferon-gamma secretion from target cells, i.e., eliminate CD161-mediated inhibition. In some embodiments, the anti-CD161 antibodies disclosed herein inhibit or reduce specific acidic sphingomyelinase (aSMase) stimulation in target cells.

[0074] The "Fc region" (fragment crystallizable region), "Fc domain," or "Fc" refers to the C-terminal region of the antibody heavy chain, mediating the binding of immunoglobulins to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or to the first component (C1q) of the classical complement system. Therefore, the Fc region includes the constant region of the antibody excluding the first constant-region immunoglobulin domain (e.g., CH1 or CL). In the antibody isotypes IgG, IgA, and IgD, the Fc region contains two identical protein fragments derived from the second (CH2) and third (CH3) constant domains of the two heavy chains of the antibody, while the Fc region of IgM and IgE contains three heavy-chain constant domains (CH domains 2-4) in each polypeptide chain. In the case of IgG, the Fc region includes the CH2 and CH3 of the immunoglobulin domain, as well as the hinge between the CH1 and CH2 domains. The definition of the boundary of the Fc region of an immunoglobulin heavy chain can vary, but as defined herein, the human IgG heavy chain Fc region is defined as extending from amino acid residue D221 in IgG1, V222 in IgG2, L221 in IgG3, and P224 in IgG4 to the carboxyl terminus of the heavy chain. Here, numbering follows the EU index based on Kabat. The CH2 domain of the human IgG Fc region extends from amino acid 237 to amino acid 340, and the CH3 domain is located on the C-terminal side of the CH2 domain within the Fc region. That is, it extends from amino acid 341 to amino acid 447 or 446 (if the C-terminal lysine residue is absent) or 445 (if the C-terminal glycine and lysine residues are absent) of IgG. As used herein, the Fc region may be a natural sequence Fc containing any allotype variant or a variant Fc (e.g., an Fc not naturally occurring).

[0075] A "natural sequence Fc region" or "natural sequence Fc" contains an amino acid sequence identical to that of a naturally occurring Fc region. Natural sequence human Fc regions include natural sequence human IgG1 Fc regions, natural sequence human IgG2 Fc regions, natural sequence human IgG3 Fc regions, and natural sequence human IgG4 Fc regions, as well as their naturally occurring variants. Natural sequence Fc includes various allotypes of Fc (see, for example, Jefferis et al. (2009) mAbs 1:1).

[0076] The term “epitope” or “antigenic determinant” refers to the site on an antigen (e.g., CD161) to which an immunoglobulin or antibody specifically binds, and is defined, for example, by the specific method used for its identification. Epitopes can be formed from both continuous amino acids (usually linear epitopes) or discontinuous amino acids juxtaposed by the folding of the protein’s tertiary structure (usually structural epitopes). Epitopes formed from continuous amino acids are typically, though not always, retained upon exposure to denaturing solvents, while epitopes formed by tertiary folding are typically lost upon processing with denaturing solvents. Epitopes typically contain at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in their unique spatial structure. Methods for determining which epitopes a given antibody binds to (i.e., epitope mapping) are well known in the art and include, for example, immunoblotting and immunoprecipitation assays. These test the reactivity of overlapping or sequential peptides (e.g., derived from CD161) with a given antibody (e.g., an anti-CD161 antibody). Methods for determining the spatial structure of epitopes include techniques in the art and those described herein, such as X-ray crystallography, X-ray cocrystallography, antigen mutation analysis, two-dimensional nuclear magnetic resonance, and HDX-MS (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, GEMorris, Ed. (1996)).

[0077] The term "epitope mapping" refers to the process of identifying molecular determinants for antibody-antigen recognition.

[0078] With respect to two or more antibodies, the term “binding to the same epitope” means that the antibodies bind to the same segment of amino acid residues determined by a given method. Techniques for determining whether an antibody binds to the “same epitope on CD161” as the antibodies described herein include epitope mapping methods such as X-ray crystallography and hydrogen / deuterium exchange mass spectrometry (HDX-MS) of antigen:antibody complexes, which provide atomic resolution of the epitope. Other methods involve monitoring the binding of an antibody to an antigen fragment or a variant of the antigen, and if binding is lost due to modification of amino acid residues in the antigen sequence, it is often considered an indicator that it is an epitope component. In addition, computer combinatorial methods for epitope mapping can also be used. These methods rely on the ability of the antibody of interest to isolate specific short peptides from a combinatorial phage display peptide library based on affinity. Antibodies having the same VH and VL or the same CDR1, 2, and 3 sequences are expected to bind to the same epitope.

[0079] An antibody that "competes with another antibody for binding to its target" refers to an antibody that (partially or completely) inhibits the binding of another antibody to its target. Whether two antibodies compete with each other for binding to their target, i.e., whether one antibody inhibits the other antibody's binding to its target and to what extent, can be determined using known competition experiments, such as BIACORE® surface plasmon resonance (SPR) analysis. In some embodiments, an antibody competes with another antibody, inhibiting binding to the target by at least 50%, 60%, 70%, 80%, 90%, or 100%. The degree of inhibition or competition may vary depending on which antibody is the "blocking antibody" (i.e., the cold antibody that is initially incubated with the target). Competitive assays can be performed as described, for example, in Ed Harlow and David Lane, Cold Spring Harbor Protoc; 2006; doi:10.1101 / pdb.prot4277 or Chapter 11 of “Using Antibodies” by Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Two antibodies are said to “cross-compete” if, in a competitive experiment, the antibodies inhibit each other bidirectionally, i.e., by at least 50%, regardless of which antibody is brought into contact with the antigen first.

[0080] Competitive binding assays to determine whether two antibodies compete or cross-compete for binding include, for example, competition for binding to CD161-expressing cells by flow cytometry, as described in the examples. Other methods include SPR (e.g., BIACORE®), solid-phase radioimmunoassay (RIA) direct or indirect, solid-phase enzyme immunoassay (EIA) direct or indirect, sandwich competitive assay (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid-phase biotin-avidin EIA direct assay (see Kirkland et al., J.Immunol. 137:3614 (1986)); solid-phase direct labeling assay, solid-phase direct labeling sandwich assay (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid-phase labeled RIA direct assay using I-125 labeling (see Morel et al., Mol.Immunol. 25(1):7 (1988)); solid-phase biotin-avidin EIA direct assay (see Cheung et al., Virology This includes 176:546(1990)); and directly labeled RIA (Moldenhauer et al., Scand. J. Immunol. 32:77(1990)).

[0081] As used herein, the terms “specific binding,” “selective binding,” “selectively binding,” and “specifically binding” refer to the binding of an antibody to an epitope on a given antigen. Typically, an antibody binds to approximately 10 cells, as determined by (i) surface plasmon resonance (SPR) technology in a BIACORE® 2000 instrument using an antibody as a ligand with a given antigen, e.g., recombinant human CD161, as an analyte, or by Scatchard analysis of antibody binding to antigen-positive cells. -7 Less than M, for example, approximately 10 -8 M, 10 -9 M or 10 -10 Equilibrium dissociation constant (K) less than or even lower than M D(ii) binds to the given antigen with an affinity at least twice as high as the affinity for binding to nonspecific antigens other than the given antigen or closely related antigen (e.g., BSA, casein). Therefore, an antibody that "specifically binds to CD161" is 10 -7 M or less, for example, approximately 10 -8 M, 10 -9 M or 10 -10 K less than M or even lower D This refers to antibodies that bind to CD161. In some embodiments, such antibodies that do not cross-react with CD161 from non-human species exhibit essentially undetectable binding to these proteins in standard binding assays.

[0082] "k assoc " or "k a The term "k" as used herein is intended to refer to the association rate of a particular antibody-antigen interaction, while "k" dis " or "k d The term "K" is intended, as used herein, to refer to the dissociation rate of a particular antibody-antigen interaction. D The term "k" as used herein is intended to refer to the dissociation constant, which is k d and k a The ratio of (i.e., k d / k a ) is obtained from and expressed as molar concentration (M). K of the antibody D The value can be determined using methods well established in the art. Antibody K D Methods available to determine this include surface plasmon resonance, biosensor systems such as the BIACORE® system, or flow cytometry and Scatchard analysis.

[0083] As used herein, the term "high affinity" for an IgG antibody means 10% affinity for the target antigen. -8 M or less, 10- 9 M or less, or 10 -10 K below M DThis refers to antibodies that possess high affinity. However, "high affinity" binding can vary depending on the antibody isotype. For example, the "high affinity" binding to the IgM isotype is 10 -10 M or less, or 10 -8 K below M D This refers to antibodies that possess [a certain characteristic].

[0084] "EC" in in vitro or in vivo assays using antibodies or their antigen-binding fragments 50 The term "maximum response" refers to the concentration of an antibody or its antigen-binding moiety that induces a response that is 50% of the maximum response, i.e., an intermediate level between the maximum response and the baseline.

[0085] As used herein, the term “naturally occurring” means, when applied to a subject, that subject can be found in nature. For example, polypeptide sequences or polynucleotide sequences found in organisms (including viruses) that can be isolated from natural sources and have not been intentionally modified in a laboratory are considered naturally occurring.

[0086] A "polypeptide" refers to a chain containing at least two consecutively linked amino acid residues, with no upper limit on chain length. One or more amino acid residues in a protein may undergo modifications, such as glycosylation, phosphorylation, or disulfide bond formation, but are not limited to these modifications. A "protein" may contain one or more polypeptides.

[0087] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. Nucleic acid molecules may be single-stranded or double-stranded, and may be cDNA.

[0088] "Conservative amino acid substitution" means that an amino acid residue is substituted with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with non-charged side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). In some embodiments, non-essential amino acid residues in the predicted anti-CD161 antibody are replaced with other amino acid residues from the same side-chain family. Methods for identifying conserved nucleotide and amino acid substitutions that do not eliminate antigen binding are well known in the art (see, for example, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al. Protein Eng. 12(10):879-884 (1999); and Burks et al. Proc. Natl. Acad. Sci. USA 94:412-417 (1997)).

[0089] In the case of nucleic acids, the term "substantial homology" indicates that two nucleic acids, or their specified sequences, are identical in at least approximately 80%, at least approximately 90%–95%, or at least approximately 98%–99.5% of nucleotides, with appropriate nucleotide insertions or deletions, when optimally aligned and compared. Alternatively, substantial homology exists when segments hybridize to the chain complement under selective hybridization conditions.

[0090] In the case of polypeptides, the term "substantial homology" indicates that two polypeptides, or their specified sequences, are identical in at least about 80%, at least about 90%–95%, or at least about 98%–99.5% of their amino acids, with appropriate amino acid insertions or deletions, when optimally aligned and compared.

[0091] The percentage of identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap (i.e., percentage of identity = number of identical positions / total number of positions × 100). Sequence comparison and determination of the percentage of identity between two sequences can be performed using mathematical algorithms, as described in the non-restrictive examples below.

[0092] The percentage of identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at worldwideweb.gcg.com) with the NWSgapdna.CMP matrix and gap weights of 40, 50, 60, 70, or 80 and length weights of 1, 2, 3, 4, 5, or 6. The percentage of identity between two nucleotide or amino acid sequences can also be determined using the E. Meyers and W. Miller algorithm (CABIOS, 4:11-17 (1989)) incorporated into the ALIGN program (version 2.0) with the PAM120 weighted residue table, gap length penalty 12, and gap penalty 4. In addition, the percentage of identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48): 444-453 (1970)) algorithm, which is incorporated into the GAP program of the GCG software package (available at http: / / www.gcg.com), using either the Blossum 62 matrix or the PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6.

[0093] The nucleic acid and protein sequences described herein can be further used, for example, as "query sequences" to perform searches in public databases to identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) described in Altschul, et al. (1990) J.Mol.Biol.215:403-10. A BLAST nucleotide search can be performed using the NBLAST program with a score of 100 and a word length of 12 to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. A BLAST protein search can be performed using the XBLAST program with a score of 50 and a word length of 3 to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. To obtain alignments including gaps for comparison purposes, Gapped BLAST can be used, as described in Altschul et al., (1997) Nucleic Acids Res.25(17):3389-3402. When using the BLAST and Gapped BLAST programs, you can use the default parameters of each program (e.g., XBLAST and NBLAST). See worldwideweb.ncbi.nlm.nih.gov for details.

[0094] Nucleic acids may exist in whole cells, in cell lysates, or in partially purified or substantially pure forms. A nucleic acid is considered "isolated" or "substantially pure" if it has been purified from other cellular components or other contaminants, e.g., other cellular nucleic acids (e.g., other parts of chromosomes) or proteins, by standard techniques including alkali / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art. See F. Ausubel, et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0095] Nucleic acids, such as cDNA, can be mutated according to standard techniques to provide gene sequences. In the case of coding sequences, these mutations can affect the amino acid sequence as desired. In particular, DNA sequences that are substantially homologous to or derived from natural V, D, J, stationary sequences, switches, and other such sequences described herein are assumed (where “derived” indicates that the sequence is identical to or modified from another sequence).

[0096] As used herein, the term “vector” is intended to mean a nucleic acid molecule capable of transporting another ligated nucleic acid. One type of vector is a “plasmid,” which refers to a circular double-stranded DNA loop to which additional DNA segments can be ligated. Another type of vector is a viral vector to which additional DNA segments can be ligated into a viral genome. Certain vectors are capable of self-replication within the host cell into which they are introduced (e.g., bacterial vectors with bacterial origins of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be incorporated into the host cell's genome upon introduction into the host cell, thereby replicating with the host genome. Furthermore, a given vector can direct the expression of a functionally ligated gene. Such vectors are referred herein as “recombinant expression vectors” (or simply “expression vectors”). Expression vectors typically used in recombinant DNA technology are often in the form of plasmids. Since plasmids are the most commonly used form of vector, “plasmid” and “vector” may be used interchangeably herein. However, other forms of expression vectors, such as viral vectors (e.g., replication-deficient retroviruses, adenoviruses, and adeno-associated viruses), are also included, and these perform equivalent functions.

[0097] As used herein, the term “recombinant host cell” (or simply “host cell”) is intended to mean a cell containing nucleic acids not naturally present in the cell, which may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terminology is intended to refer not only to a specific target cell but also to the offspring of such cell. Because certain modifications may occur in subsequent generations due to either mutation or environmental influences, such offspring may not actually be identical to the parent cell, but they still fall within the scope of the term “host cell” as used herein.

[0098] As used herein, “administer” means physically introducing a composition containing a therapeutic agent into a target using any of the various methods and delivery systems known to those skilled in the art. Various routes of administration of anti-CD161 antibodies described herein include, for example, intravenous, intramuscular, subcutaneous, intraperitoneal, intravesical, transdermal, intraspinal, or other parenteral routes of administration by injection or infusion. In some embodiments, administration includes parenteral administration. As used herein, “parenteral administration” means a mode of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intraocular, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. Alternatively, the antibodies described herein may be administered via parenteral routes such as local, dermal, or mucosal administration, for example, intranasal, oral, vaginal, rectal, sublingual, or local administration. Administration may also be performed, for example, once, multiple times, and / or over a longer period of time.

[0099] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to any type of intervention or process or administration of an active agent performed on a subject with the aim of reversing, mitigating, improving, inhibiting, slowing or preventing the progression, onset, severity or recurrence of disease-related symptoms, complications, conditions, or biochemical indicators, or increasing overall survival. Treatment may be performed on a subject with the disease or a subject without the disease (for example, for prevention).

[0100] The term “effective dose” or “effective dosage” is defined as the amount sufficient to achieve, or at least partially achieve, the desired effect. A “therapeutically effective dose” or “therapeutic dosage” of a drug or treatment is any amount of the drug, used alone or in combination with another treatment, that promotes disease regression, as demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods, an increase in overall survival (the length of time a patient diagnosed with a disease, such as cancer, survives from either the date of diagnosis or the start of treatment for the disease), or the prevention of functional impairment or disability due to the distress of the disease. A therapeutically effective dose or dosage of a drug also includes a “prophylactically effective dose” or “prophylactically effective dosage,” which is any amount of the drug, used alone or in combination with another treatment, that inhibits the onset or recurrence of the disease when administered to a subject at risk of developing the disease or at risk of disease recurrence. The ability of therapeutic agents to promote disease regression or inhibit disease onset or recurrence can be evaluated using a variety of methods known to experienced practitioners, such as in human subjects during clinical trials, in animal model systems to predict efficacy in humans, or by assaying drug activity in in vitro assays.

[0101] The term "patient" includes human and other mammalian subjects receiving either preventive or therapeutic treatment.

[0102] As used herein, the term “subject” includes any human or non-human animal. For example, the methods and compositions described herein can be used to treat a subject having cancer. The term “non-human animal” includes all vertebrates, e.g., mammals and non-mammals, e.g., non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0103] As used herein, the terms "ug" and "uM" are interchangeable with "μg" and "μM," respectively.

[0104] The various embodiments described herein are described in more detail in the following subsections.

[0105] II. Compositions of the Disclosure This disclosure relates to antibodies that specifically bind to CD161 and their antigen-binding moieties. In some embodiments, the anti-CD161 antibodies and their antigen-binding moieties described herein block or reduce the interaction between CD161 and CLEC2D. The binding of CD161 to CLEC2D inhibits T cell activation. Therefore, in some embodiments, the anti-CD161 antibodies and their antigen-binding moieties described herein can block or reduce the inhibition of T cell activation induced by CD161-CLEC2D while enhancing the immune response in human subjects.

[0106] Any method can be used to determine the ability of anti-CD161 to overcome the inhibition of T cell activation mediated by the CD161-CLEC2D interaction. The anti-CD161 antibodies described herein induce or enhance cytokine production by immune cells, for example, as determined by a cytokine assay. In some embodiments, the cytokine assay determines the amount of at least one cytokine secreted from immune cells contacted with the anti-CD161 antibody, where an increase in the amount of at least one cytokine indicates induction or enhancement of cytokine production by the anti-CD161 antibody.

[0107] In some embodiments, the increase in cytokine production is at least 1.5-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-fold higher compared to a control antibody (e.g., an equivalent antibody isotype that does not bind to CD161, e.g., an antibody that does not induce cytokine production). The amount of at least one cytokine produced by immune cells is compared to the amount secreted by reference immune cells, where the reference immune cells are in contact with a reference antibody. If the amount of at least one cytokine produced by immune cells is increased compared to that of reference immune cells, it indicates that cytokine production was induced or enhanced due to the blockade of the CD161-CLEC2D interaction. In some embodiments, the immune cells are either CD8+ T cells or CD4+ T cells.

[0108] In some embodiments, anti-CD161 antibodies and their antigen-binding moieties induce or enhance cytokine production by immune cells. In some embodiments, cytokine production by immune cells contacted with anti-CD161 antibodies and their antigen-binding moieties increases by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to immune cells before contact or immune cells contacted with a control antibody (e.g., an equivalent antibody isotype that does not bind to CD161). In some embodiments, the immune cells are T cells. In some embodiments, the T cells are CD8 + T cells or CD4 + These are T cells. In some cases, the immune cells are NK cells.

[0109] In some embodiments, contact between immune cells and the anti-CD161 antibody and its antigen-binding moiety described herein increases the expression of interleukin-2 (IL-2) by the immune cells. In some embodiments, IL-2 expression increases by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to IL-2 expression by immune cells before contact or by immune cells contacted with a control antibody.

[0110] In some embodiments, contact between immune cells and the anti-CD161 antibody and its antigen-binding moiety described herein increases the expression of interferon-gamma (IFNg) by the immune cells. In some embodiments, IFNg expression increases by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to IFNg expression by immune cells before contact or by immune cells contacted with a control antibody.

[0111] In some embodiments, contact between immune cells and the anti-CD161 antibody and its antigen-binding moiety described herein increases the expression of tumor necrosis factor-alpha (TNF-α) by the immune cells. In some embodiments, TNF-α expression increases by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to TNF-α expression by immune cells before contact or by immune cells contacted with a control antibody.

[0112] In some embodiments, the anti-CD161 antibody and its antigen-binding moiety described herein have high affinity for human CD161, for example, 10 -6 M or less, 10 -7 M or less, 10 -8 M or less, 10 -9 M or less, 10-10 M or less, 10 -11 M or less, 10 -12 M or less, 10 -12 M~10 -7 M, 10 -11 M~10 -7 M, 10 -10 M~10 -7 M, or 10 -9 M~10 -7 M's K D It binds to human CD161, for example by surface plasmon resonance, for example, when determined using Biacore®. -6 M or less, 10 -7 M or less, 10 -8 M or less, 10 -9 M (1nM) or less, 10 -10 M or less, 10 -12 M~10 -7 M, 10 -11 M~10 -7 M, 10 -10 M~10 -7 M, 10 -9 M~10 -7 M, or 10 -8 M~10 -7 M's K D They are joined together.

[0113] II.A. Anti-CD161 antibody In some embodiments, the antibody or its antigen-binding moiety comprises a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region (VH), the light chain comprising a light chain variable region (VL), the VH comprising VH complementarity-determining region 1 (VH-CDR1), VH-CDR2, and VH-CDR3, and the VL comprising VL-CDR1, VL-CDR2, and VL-CDR3, and VH-CDR3 This includes an amino acid sequence selected from the sequences described in SEQ ID NOs: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323, and 333. In some embodiments, VH-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, and 332. In some embodiments, VH-CDR1 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, and 331. In some embodiments, VL-CDR3 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, and 336.In some embodiments, VL-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, and 335. In some embodiments, VL-CDR1 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, and 334. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11]

[0114] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0115] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0116] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0117] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0118] In some embodiments, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0119] In some embodiments, the antibody or its antigen-binding portion is bound to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0120] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0121] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0122] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0123] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0124] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0125] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0126] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0127] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0128] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0129] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0130] In some embodiments, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0131] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0132] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0133] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0134] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0135] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0136] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0137] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0138] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0139] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0140] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0141] In some embodiments, the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0142] In some embodiments, the antibody or its antigen-binding moiety cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0143] In some embodiments, the antibody or its antigen-binding moiety binds to the same human CD161 epitope as the reference antibody, wherein the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0144] In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, and 337. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 7. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 97. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 117. In some embodiments, the antibody or its antigen-binding portion includes a VH having an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 147.In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 217. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 227. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 277. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 287. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 297. In some embodiments, the antibody or its antigen-binding portion includes a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 307.

[0145] In some embodiments, the antibody or its antigen-binding moiety includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 98. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 118. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 148.In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 218. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 228. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 278. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 288. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 298. In some embodiments, the antibody or its antigen-binding portion includes a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 308.

[0146] In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, and 337. In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising the amino acid sequence described in SEQ ID NO: 7. In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising the amino acid sequence described in SEQ ID NO: 97. In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising the amino acid sequence described in SEQ ID NO: 117. In some embodiments, the antibody or its antigen-binding moiety includes a VH comprising the amino acid sequence described in SEQ ID NO: 147. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 217. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 227. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 277. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 287. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 297. In some embodiments, the antibody or its antigen-binding portion includes a VH having the amino acid sequence described in SEQ ID NO: 307.

[0147] In some embodiments, the antibody or its antigen-binding moiety includes a VL containing an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338. In some embodiments, the antibody or its antigen-binding moiety includes a VL containing the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the antibody or its antigen-binding moiety includes a VL containing the amino acid sequence described in SEQ ID NO: 98. In some embodiments, the antibody or its antigen-binding moiety includes a VL containing the amino acid sequence described in SEQ ID NO: 118. In some embodiments, the antibody or its antigen-binding moiety includes a VL containing the amino acid sequence described in SEQ ID NO: 148. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 218. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 228. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 278. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 288. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 298. In some embodiments, the antibody or its antigen-binding portion includes a VL containing the amino acid sequence described in SEQ ID NO: 308.

[0148] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 7, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected to be described in SEQ ID NO: 8.

[0149] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 7, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, with respect to the amino acid sequence selected for description in SEQ ID NO: 8. The antibody comprises a VL containing an amino acid sequence having at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody comprises VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0150] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 7, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 8. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0151] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 7, and (ii) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, or at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 8. The antibody comprises a VL containing an amino acid sequence having sequence identity of %, at least about 98%, or at least about 99%, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6.

[0152] In some embodiments, the antibody or its antigen-binding portion includes (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 97, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 98.

[0153] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 97, and (ii) an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, and The antibody comprises a VL containing an amino acid sequence having at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0154] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 97, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 98. The antibody contains a VL containing an amino acid sequence with at least 99% sequence identity, and the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161. The reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0155] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 97, and (ii) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 98. The antibody comprises a VL containing an amino acid sequence having at least approximately 98% or at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96.

[0156] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 117, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 118.

[0157] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 117, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 118. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0158] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 117, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0159] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 117, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 118. The antibody comprises a VL containing an amino acid sequence having at least approximately 98%, or at least approximately 99%, sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116.

[0160] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 147, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected to be described in SEQ ID NO: 148.

[0161] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 147, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 148. The antibody contains a VL containing an amino acid sequence having at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0162] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 147, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0163] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 147, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 148. The antibody comprises a VL containing an amino acid sequence having at least approximately 98% or at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146.

[0164] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 217, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 218.

[0165] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 217, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 218. The antibody contains a VL containing an amino acid sequence having at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0166] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 217, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0167] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 217, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or at least about 97% sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 218. The antibody comprises a VL containing an amino acid sequence having at least approximately 98% or at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216.

[0168] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 227, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 228.

[0169] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 227, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 228. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0170] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 227, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0171] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 227, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 228. The antibody comprises a VL containing an amino acid sequence having at least approximately 98%, or at least approximately 99%, sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226.

[0172] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 277, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 278.

[0173] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 277, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 278. The antibody contains a VL containing an amino acid sequence having at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0174] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 277, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0175] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 277, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or less with respect to the amino acid sequence selectively described in SEQ ID NO: 278. The antibody comprises a VL containing an amino acid sequence having at least approximately 98% or at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276.

[0176] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 287, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 288.

[0177] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 287, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 288. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0178] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 287, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0179] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 287, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or at least about 97% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 288. The antibody comprises a VL containing an amino acid sequence having at least approximately 98%, or at least approximately 99%, sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286.

[0180] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 297, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected to be described in SEQ ID NO: 298.

[0181] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 297, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 298. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0182] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 297, and (ii) an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0183] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 297, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or less with respect to the amino acid sequence selected for description in SEQ ID NO: 298. The antibody comprises a VL containing an amino acid sequence having at least approximately 98% or at least approximately 99% sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296.

[0184] In some embodiments, the antibody or its antigen-binding portion comprises (i) a VH containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 307, and (ii) a VL containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence selected for description in SEQ ID NO: 308.

[0185] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 307, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or less sequence identity with respect to the amino acid sequence selected for SEQ ID NO: 308. The antibody contains a VL containing an amino acid sequence having at least 99% sequence identity, wherein the antibody or its antigen-binding portion is bound to the same epitope as the reference antibody, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0186] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 307, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about The antibody comprises a VL containing an amino acid sequence having 99% sequence identity, wherein the antibody or its antigen-binding portion cross-competes with the reference antibody for binding to human CD161, and the reference antibody includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0187] In some embodiments, the antibody or its antigen-binding moiety comprises (i) a VH having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO: 307, and (ii) at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, or less with respect to the amino acid sequence selected for description in SEQ ID NO: 308. The antibody comprises a VL containing an amino acid sequence having at least approximately 98%, or at least approximately 99%, sequence identity, wherein the antibody or its antigen-binding portion includes VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306.

[0188] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 7 and a VL containing the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 7 and a VL containing the amino acid sequence described in SEQ ID NO: 8. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 7 and a VL containing the amino acid sequence described in SEQ ID NO: 8.

[0189] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 97 and a VL containing the amino acid sequence described in SEQ ID NO: 98. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 97 and a VL containing the amino acid sequence described in SEQ ID NO: 98. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 97 and a VL containing the amino acid sequence described in SEQ ID NO: 98.

[0190] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 117 and a VL containing the amino acid sequence described in SEQ ID NO: 118. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 117 and a VL containing the amino acid sequence described in SEQ ID NO: 118. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 117 and a VL containing the amino acid sequence described in SEQ ID NO: 118.

[0191] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 147 and a VL containing the amino acid sequence described in SEQ ID NO: 148. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 147 and a VL containing the amino acid sequence described in SEQ ID NO: 148. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 147 and a VL containing the amino acid sequence described in SEQ ID NO: 148.

[0192] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 217 and a VL containing the amino acid sequence described in SEQ ID NO: 218. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 217 and a VL containing the amino acid sequence described in SEQ ID NO: 218. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 217 and a VL containing the amino acid sequence described in SEQ ID NO: 218.

[0193] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 227 and a VL containing the amino acid sequence described in SEQ ID NO: 228. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 227 and a VL containing the amino acid sequence described in SEQ ID NO: 228. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 227 and a VL containing the amino acid sequence described in SEQ ID NO: 228.

[0194] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 277 and a VL containing the amino acid sequence described in SEQ ID NO: 278. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 277 and a VL containing the amino acid sequence described in SEQ ID NO: 278. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 277 and a VL containing the amino acid sequence described in SEQ ID NO: 278.

[0195] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 287 and a VL containing the amino acid sequence described in SEQ ID NO: 288. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 287 and a VL containing the amino acid sequence described in SEQ ID NO: 288. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 287 and a VL containing the amino acid sequence described in SEQ ID NO: 288.

[0196] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 297 and a VL containing the amino acid sequence described in SEQ ID NO: 298. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 297 and a VL containing the amino acid sequence described in SEQ ID NO: 298. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 297 and a VL containing the amino acid sequence described in SEQ ID NO: 298.

[0197] In some embodiments, the antibody or its antigen-binding moiety includes a VH containing the amino acid sequence described in SEQ ID NO: 307 and a VL containing the amino acid sequence described in SEQ ID NO: 308. In some embodiments, the antibody or its antigen-binding moiety binds to the same epitope as the reference antibody, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 307 and a VL containing the amino acid sequence described in SEQ ID NO: 308. In some embodiments, the antibody or its antigen-binding moiety cross-competes with the reference antibody for binding to human CD161, where the reference antibody includes a VH containing the amino acid sequence described in SEQ ID NO: 307 and a VL containing the amino acid sequence described in SEQ ID NO: 308.

[0198] In some embodiments, the antibody or its antigen-binding moiety includes one or more post-translational modifications. In some embodiments, the antibody or its antigen-binding moiety includes one or more post-translational modifications that increase the in vivo half-life of the antibody or its antigen-binding moiety. In some embodiments, the antibody or its antigen-binding moiety is PEGylated.

[0199] In some embodiments, the antigen-binding portion of the antibody includes a VHH, vNAR, microbody, nanobody, scFv, or any combination thereof.

[0200] II.B. Bispecific and multispecific antibodies The anti-CD161 antibodies and their antigen-binding moieties described herein can be used to form bispecific and multispecific molecules. The anti-CD161 antibody or its antigen-binding moiety can be derivatized or ligated to another functional molecule, such as another peptide or protein (e.g., another antibody or receptor ligand), to produce a bispecific molecule that binds to at least two different binding sites or target molecules. For example, the anti-CD161 antibody can be ligated to an antibody or scFv that specifically binds to a tumor antigen. In fact, the antibodies described herein can be derivatized or ligated to two or more other functional molecules to produce multispecific molecules that bind to three or more different binding sites and / or target molecules. To produce the bispecific molecules described herein, the antibodies described herein can be functionally ligated (e.g., by chemical coupling, gene fusion, non-covalent bonding, or other methods) to one or more other binding molecules, such as another antibody, antibody fragment, peptide, or binding mimetic.

[0201] Accordingly, this specification provides a bispecific molecule comprising at least one first binding specificity to CD161 and a second binding specificity to a second target epitope. Furthermore, this specification provides a multispecific molecule comprising at least one first binding specificity to CD161, a second binding specificity to a second target epitope, and a third binding specificity to a third target epitope.

[0202] In some embodiments, the bispecific and multispecific molecules described herein include, as binding specificity, at least one antibody or a fragment of that antibody (e.g., Fab, Fab', F(ab')2, Fv, or single-chain Fv(scFv)). The antibody may also be a light-chain or heavy-chain dimer, or any smallest fragment thereof, such as Fv or a single-chain construct as described in U.S. Patent No. 4,946,778 by Ladner et al.

[0203] Human monoclonal antibodies are preferred, but other antibodies that can be used in the bispecific and multispecific molecules described herein include mouse monoclonal antibodies, chimeric monoclonal antibodies, and humanized monoclonal antibodies.

[0204] The bispecific and multispecific molecules described herein can be prepared by conjugating their binding specificity components using methods known in the art. For example, each binding specificity component of a bispecific or multispecific molecule can be prepared separately and then conjugated together. When the binding specificity component is a protein or peptide, various coupling agents or crosslinking agents can be used for covalent conjugation. Examples of crosslinking agents include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohaxane-1-carboxylate (sulfo-SMCC) (see, for example, Karpovsky et al. (1984) J. Exp. Med. 160:1686; Liu, MA et al. (1985) Proc. Natl. Acad. Sci. USA 82:8648). Other methods include Paulus (1985) Behring Ins. Mitt. No. 78, 118-132; Brennan et al. (1985) Science 229: 81-83), and Glennie et al. (1987) J. Immunol. 139: 2367-2375). Some conjugates are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).

[0205] When the binding specificity moiety is an antibody, they can be conjugated via sulfhydryl linkages in the C-terminal hinge regions of the two heavy chains. In some embodiments, the hinge region is modified to contain an odd number of sulfhydryl residues (preferably one) prior to conjugation.

[0206] Alternatively, multiple binding specificity moieties can be encoded in the same vector, expressed in the same host cell, and assembled. This method is particularly useful when the bispecific molecule is an mAb×mAb, mAb×Fab, mAb×(scFv)2, Fab×F(ab')2, or ligand×Fab fusion protein. The bispecific antibody may contain an antibody with scFv at the C-terminus of each heavy chain. The bispecific molecules described herein may be a single-chain molecule containing one single-chain antibody and a binding determinant, or a single-chain bispecific molecule containing two binding determinants. The bispecific molecule may contain at least two single-chain molecules. Methods for preparing bispecific molecules are described, for example, in U.S. Patents 5,260,203, 5,455,030, 4,881,175, 5,132,405, 5,091,513, 5,476,786, 5,013,653, 5,258,498, and 5,482,858.

[0207] The binding of bispecific and multispecific molecules to their specific targets can be confirmed using methods recognized in the art, such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassays (e.g., growth inhibition), or Western blot assays. Each of these assays generally detects the presence of a particular protein-antibody complex by using a labeling reagent (e.g., antibody) specific to the complex in question.

[0208] In some embodiments, the bispecific antibody comprises (i) an anti-CD161 antibody or its antigen-binding moiety as disclosed herein and (ii) an antibody or its antigen-binding moiety that specifically binds to a tumor antigen. In some embodiments, the tumor antigen is CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD70, CD171, CD33, EGFRvIII, GD2, GD3, TnAg, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, SSEA-4, CD20, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5 HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WTl, NY-ESO-1, LAGE-la, MAGE-Al, Regmine, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 variant, prostain, surviving, telomerase, PCTA-1 / galectin 8, Melan A / MARTl, Ras variant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2)Selected from ETS fusion genes, NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mutant hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, extracellular components of APRIL protein, or any combination thereof. In some embodiments, TCR is AFP, CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, TnAg, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, Mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, SSEA-4, CD20, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5 HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WTl, NY-ESO-1, LAGE-la, MAGE-Al, Regmine, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 variant, prostain, surviving, telomerase, PCTA-1 / galectin 8, melan A / MARTl, Ras variant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2) The target is the extracellular component of the ETS fusion gene, NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mutant hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, APRIL protein, and any combination thereof. In some embodiments, the bispecific antibody comprises (i) an anti-CD161 antibody or its antigen-binding moiety disclosed herein, and (ii) CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD70, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, SSEA-4, CD20, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5 HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WTl, NY-ESO-1, LAGE-la, MAGE-Al, Regmine, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 variant, prostain, surviving, telomerase, PCTA-1 / galectin 8, Melan A / MARTl, Ras variant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2) The antibody comprises an antibody or its antigen-binding moiety that specifically binds to a tumor antigen selected from the following: ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mutant hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, the extracellular portion of the APRIL protein, or any combination thereof. In some embodiments, TCR is AFP, CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, TnAg, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, Mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, SSEA-4, CD20, Folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, Tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5 HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WTl, NY-ESO-1, LAGE-la, MAGE-Al, Regmine, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 variant, prostain, surviving, telomerase, PCTA-1 / galectin 8, Melan A / MARTl, Ras variant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2) The target is the extracellular component of the ETS fusion gene, NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mutant hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, APRIL protein, and any combination thereof.

[0209] II.C. Nucleic acid molecules Another aspect described herein relates to a nucleic acid molecule encoding the anti-CD161 antibody described herein. The nucleic acid may exist in whole cells, in cell lysates, or in a partially purified or substantially pure form. A nucleic acid is considered "isolated" or "substantially pure" if it has been purified from other cellular components or other contaminants, e.g., other cellular nucleic acids (e.g., other chromosomal DNA, e.g., chromosomal DNA linked to naturally isolated DNA) or proteins, by standard techniques including alkali / SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis, and others well known in the art. See F. Ausubel, et al., ed. (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. The nucleic acids described herein may be, for example, DNA or RNA, and may or may not contain intron sequences. In some aspects, the nucleic acid is a cDNA molecule.

[0210] The nucleic acids described herein can be obtained using standard molecular biological techniques. For antibodies expressed by hybridomas (e.g., hybridomas prepared from transgenic mice possessing the human immunoglobulin genes further described below), the cDNA encoding the light and heavy chains of the antibody produced by the hybridoma can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from immunoglobulin gene libraries (e.g., those using phage display techniques), the nucleic acids encoding the antibody can be recovered from the library.

[0211] Some nucleic acid molecules described herein encode the VH and VL sequences of the anti-CD161 antibodies disclosed herein.

[0212] The nucleic acid molecules of this disclosure may be modified to remove specific sequences, such as restriction enzyme recognition sequences, or to optimize codons.

[0213] A method for producing an anti-CD161 antibody disclosed herein may involve expressing the heavy and light chains in a cell line containing nucleotide sequences encoding the heavy and light chains together with a signal peptide. Host cells containing these nucleotide sequences are included herein.

[0214] Once DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated using standard recombinant DNA techniques to, for example, change the variable region gene into a full-length antibody chain gene, a Fab fragment gene, or an scFv gene. In these manipulations, the DNA fragment encoding VL or VH is functionally ligated to another DNA fragment encoding another protein, such as an antibody constant region or a flexible linker. As used in this context, the term "functionally ligated" is intended to mean that the two DNA fragments are ligated in such a way that the amino acid sequences encoded by the two DNA fragments remain in frame.

[0215] Isolated DNA encoding the VH region can be converted into a full-length heavy chain gene by functionally ligating the VH-encoding DNA to another DNA molecule encoding the heavy chain constant region (hinge, CH1, CH2, and / or CH3). The sequences of human heavy chain constant region genes are known in the art (see, for example, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments containing these regions can be obtained by standard PCR amplification. The heavy chain constant region may be the IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region, for example, the IgG1 region. In the case of Fab fragment heavy chain genes, the VH-encoding DNA can be functionally ligated to another DNA molecule encoding only the heavy chain CH1 constant region.

[0216] Isolated DNA encoding the VL region can be converted into full-length light chain genes (and Fab light chain genes) by functionally ligating the VL-encoding DNA to another DNA molecule encoding the light chain constant region CL. The sequences of human light chain constant region genes are known in the art (see, for example, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments containing these regions can be obtained by standard PCR amplification. The light chain constant region may be a kappa or lambda constant region.

[0217] To construct the scFv gene, DNA fragments encoding VH and VL are functionally ligated to another fragment encoding a flexible linker, such as another fragment encoding the amino acid sequence (Gly4-Ser)3. This allows the VH and VL sequences to be expressed as a continuous single-chain protein, with the VL and VH regions linked by a flexible linker.

[0218] Some aspects of this disclosure relate to vectors or sets of vectors comprising nucleic acid molecules disclosed herein. In some aspects, the vector is a viral vector. In some aspects, the vector is a viral particle or virus. In some aspects, the vector is a mammalian vector. In some aspects, the vector is a bacterial vector.

[0219] In certain embodiments, the vector is a retroviral vector. In some embodiments, the vector is selected from adenovirus vectors, lentiviruses, Sendai viruses, baculovirus vectors, Epstein-Barr virus vectors, papova virus vectors, vaccinia virus vectors, herpes simplex virus vectors, and adeno-associated virus (AAV) vectors. In certain embodiments, the vector is an AAV vector. In some embodiments, the vector is a lentivirus. In certain embodiments, the vector is an AAV vector. In some embodiments, the vector is a Sendai virus. In some embodiments, the vector is a hybrid vector. Examples of hybrid vectors that can be used in this disclosure can be found in Huang and Kamihira, Biotechnol. Adv. 31(2):208-23(2103), which is incorporated herein by reference in its entirety.

[0220] Some aspects of this disclosure relate to host cells containing nucleic acid molecules, sets of nucleic acid molecules, vectors, or sets of vectors disclosed herein. In some aspects, the host cell is a mammalian cell. In some aspects, the host cell is an in vitro cell.

[0221] II.D. Pharmaceutical Compositions Furthermore, pharmaceutical compositions are also provided, for example, comprising an anti-CD161 antibody, nucleic acid molecule, vector, or host cell disclosed herein, and one or more pharmaceutically acceptable carriers.

[0222] In some embodiments, the composition further comprises a filler. The filler may be selected from the group consisting of NaCl, mannitol, glycine, alanine, and any combination thereof. In another embodiment, the composition comprises a stabilizer. The stabilizer may be selected from the group consisting of sucrose, trehalose, raffinose, arginine, or any combination thereof. In yet another embodiment, the composition comprises a surfactant. In some embodiments, the surfactant is selected from polysorbate 80 (PS80), polysorbate 20 (PS20), and any combination thereof. In certain embodiments, the composition further comprises a chelating agent. In some embodiments, the chelating agent is selected from diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid, nitrilotriacetic acid, and any combination thereof. In some embodiments, the composition further comprises NaCl, mannitol, pentetic acid (DTPA), sucrose, PS80, or any combination thereof.

[0223] As used herein, “pharmaceutically acceptable carrier” includes any physiologically compatible solvent, dispersion medium, coating agent, antimicrobial and antifungal agent, isotonic agent and absorption retarder, etc. In some embodiments, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, intrathecal, or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., antibody, immunoconjugate, or bispecific component, may be coated with a material that protects the compound from the action of acids and other natural conditions that may inactivate the compound.

[0224] The pharmaceutical compounds described herein may include one or more pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" refer to salts that retain the desired biological activity of the parent compound and do not impart any undesirable toxic effects (see, e.g., Berge, S.M., et al. (1977) J. Pharm. Sci. 66:1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphorous acid, and those derived from non-toxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Base addition salts include those derived from alkaline earth metals such as sodium, potassium, magnesium, calcium, and those derived from non-toxic organic amines such as Ν,Ν'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine, etc.

[0225] The pharmaceutical compositions described herein may also include pharmaceutically acceptable antioxidants. Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium dithionite, sodium sulfite, etc., (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc., and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0226] Examples of suitable aqueous and non-aqueous carriers that can be employed in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Appropriate fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0227] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the presence of microorganisms can be ensured by the above sterilization procedures or by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include in the composition isotonic agents such as sugars, sodium chloride, etc. In addition, sustained absorption of injectable pharmaceutical forms can be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0228] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is known in the art. The use thereof in the pharmaceutical compositions described herein is contemplated, except where any conventional media or agent is incompatible with the active compound. The pharmaceutical compositions may or may not contain a preservative. Auxiliary active compounds can be incorporated into the compositions.

[0229] Therapeutic compositions must typically be sterile and stable under manufacturing and storage conditions. Compositions can be formulated as solutions, microemulsions, liposomes, or other ordered structures suitable for high drug concentrations. Carriers may be solvents or dispersion media containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Adequate fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Often, compositions may contain isotonic agents, such as sugars, polyalcohols such as mannitol and sorbitol, or sodium chloride. Sustained absorption of injectable compositions can be achieved by including absorption-delaying agents, such as monostearate and gelatin, in the composition.

[0230] Sterile solutions for injection can be prepared by adding the required amount of the active compound to a suitable solvent, along with one or a combination of the components listed above as needed, and then performing sterile microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other necessary components from those listed herein. In the case of sterile powders for the preparation of sterile solutions for injection, some preparation methods include vacuum drying and freeze-drying, which yield a powder containing the active ingredient and any desired additional components from a pre-sterile filtered solution.

[0231] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form may vary depending on the target being treated and the specific mode of administration. Generally, this amount is the amount of the composition that produces the therapeutic effect. Generally, this amount can range from approximately 0.01 percent to approximately 99 percent of the active ingredient, approximately 0.1 percent to approximately 70 percent, or approximately 1 percent to approximately 30 percent of the active ingredient, in combination with pharmaceutically acceptable carriers.

[0232] The compositions described herein may be administered via one or more routes of administration using various methods known in the art. As will be understood by those skilled in the art, the route of administration and / or mode of administration may vary depending on the desired outcome. The routes of administration of the anti-CD161 antibodies described herein may include, for example, intravenous, intramuscular, subcutaneous, intraperitoneal, intraspinal, or other parenteral routes of administration by injection or infusion.

[0233] The active compound can be prepared with a carrier that prevents the rapid release of the compound, such as in controlled-release formulations including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods for preparing such formulations are patented or commonly known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0234] III. Method of Disclosure III.A. Treatment method Some aspects of this disclosure relate to methods for treating a disease or disorder in a subject, comprising administering to the subject an anti-CD161 antibody disclosed herein, a polynucleotide encoding an anti-CD161 antibody, a vector comprising a polynucleotide, a host cell comprising a polynucleotide, or any combination thereof. In some aspects, the disease or disorder includes cancer.

[0235] Some aspects of this disclosure relate to a method for treating cancer in a subject requiring treatment, comprising administering to the subject an effective dose of a composition disclosed herein (e.g., an antibody, polynucleotide, vector, host cell, or pharmaceutical composition). Other aspects of this disclosure relate to a method for killing tumor cells in a subject requiring treatment, comprising administering to the subject an effective dose of a composition disclosed herein. Other aspects of this disclosure relate to a method for reducing tumor size in a subject requiring treatment, comprising administering to the subject an effective dose of a composition disclosed herein. Other aspects of this disclosure relate to inhibiting tumor metastasis in a subject requiring treatment, comprising administering to the subject an effective dose of a composition disclosed herein. In some aspects, the subject is a human.

[0236] Some aspects of this disclosure relate to methods for inducing an immune response in a subject requiring such induction, comprising administering to the subject an anti-CD161 antibody disclosed herein, a polynucleotide encoding an anti-CD161 antibody, a vector containing a polynucleotide, a host cell containing a polynucleotide, or any combination thereof. In some aspects, the subject is suffering from cancer.

[0237] Some aspects of this disclosure relate to methods for activating immune cells, comprising contacting the immune cells with an anti-CD161 antibody disclosed herein, a polynucleotide encoding an anti-CD161 antibody, a vector comprising a polynucleotide, a host cell comprising a polynucleotide, or any combination thereof. In some aspects, the immune cells are contacted in vitro. In some aspects, the immune cells are contacted ex vivo. In some aspects, the immune cells are present in a human subject; that is, the immune cells are contacted in vivo. In some aspects, the subject is cancerous.

[0238] In some embodiments, cancer includes solid tumors. In some embodiments, cancer includes hematological malignancies. In some embodiments, cancer is locally advanced. In some embodiments, cancer is metastatic. In some embodiments, the tumor is recurrent. In some embodiments, the tumor is refractory. In some embodiments, the tumor is recurrent and / or refractory after one or more prior treatments for treating the tumor. In some embodiments, one or more prior treatments include standard treatment. In some embodiments, one or more prior treatments include chemotherapy. In some embodiments, one or more prior treatments include immunotherapy. In some embodiments, one or more prior treatments include surgery. In some embodiments, one or more prior treatments include radiotherapy.

[0239] In some aspects, cancer includes acoustic neuroma, acute lymphoblastic leukemia, acute myeloid leukemia, adenocarcinoma, and cancers of the urinary tract, as well as carcinomas, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, biliary tract cancer, bladder cancer, bone cancer, brain cancer, brainstem glioma, breast cancer, bronchogenic carcinoma, Burkitt lymphoma and marginal zone B-cell lymphoma, adrenal cancer, anal cancer, gastrointestinal cancer, endocrine cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical carcinoma, endometrial carcinoma, fallopian tube carcinoma, renal pelvis carcinoma, vaginal carcinoma, vulvar carcinoma, and central nervous system CNS cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, craniopharyngioma, cystadenocarcinoma, fetal cancer, endometrial cancer, endosarcoma, environmentally induced cancer (including those induced by asbestos), ependymoma, epidermal carcinoma, epithelial carcinoma, esophageal cancer, esophageal carcinoma, Ewing's tumor, ocular cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, germ cell tumor, glioblastoma (e.g., glioblastoma multiforme), glioma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatoma, Hodgkin's disease, carcinoma in situ, Kaposi's sarcoma, kidney cancer (e.g.) Renal cell carcinoma (RCC), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung carcinoma, intralymphatic sarcoma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyeloblastic myelomonocytic monocytic erythroleukemia, myxosarcoma, nasopharyngeal cancer, neoplasms of the central nervous system (CNS), neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer (NSCLC), non-small cell lung carcinoma, oligodendroglioma, oral cancer (e.g., lips, tongue, mouth and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary carcinoma, papillary carcinoma, pediatric sarcoma, pineal gland Tumors, pituitary adenoma, polycythemia vera lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate cancer), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, soft tissue sarcoma, sebaceous gland carcinoma, seminomas, nasal cavity / sinus natural killer cancers, skin cancer, small cell lung cancer (SCLC), pediatric solid tumors, spinal axial tumors, squamous cell carcinoma, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synoviomas, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-associated cancer or cancer of virus origin (e.g., human papillomavirus (HPV-associated or HPV-derived tumors)),Waldenström macroglobulinemia and Wilms' tumor; as well as any combination of the aforementioned cancers.

[0240] In some embodiments, the control suffers from an infectious disease; that is, in some embodiments, the disease or disorder includes infectious diseases. In some embodiments, the infectious disease is selected from bacterial infections, fungal infections, viral infections, parasitic infections, or any combination thereof. In some embodiments, the infectious disease includes infections caused by influenza, herpes, giardia, malaria, leishmania, or any combination thereof. In some embodiments, the infectious disease includes infections caused by human immunodeficiency virus (HIV), hepatitis virus herpesvirus, adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus, or arbovirus encephalitis virus, or any combination thereof. In some embodiments, infectious diseases include infections caused by Chlamydia, Rickettsia, Mycobacteria, Staphylococcus, Streptococcus, Pneumococcus, Meningococcus, Conococcus, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacillus, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease, or any combination thereof. In some embodiments, infectious diseases include infections caused by Candida, Cryptococcus neoformans, Aspergillus, Mucorales, Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, or Histoplasma capsulatum, or any combination thereof.In some embodiments, infectious diseases include infections caused by Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, or Nippostrongylus brasiliensis, or any combination thereof.

[0241] In some cases, the subjects suffer from autoimmune diseases.

[0242] The compositions of this disclosure can be administered using any pharmaceutically acceptable route. In some embodiments, the compositions (e.g., antibodies, polynucleotides, vectors, host cells, or pharmaceutical compositions) are administered intravenously, intraperitoneally, intramuscularly, intraarterially, intrathecally, lymphatically, intrafocally, intracapsularly, intraorbitally, intracardiacly, intradermally, transtracheally, subcutaneously, subepidermally, intraarticularly, subcapsularly, subarachnoidally, intraspinally, epidurally, intrasternally, topically, epidermally, mucous membranely, or any combination thereof. In some embodiments, the compositions are administered intravenously. In some embodiments, the compositions are administered subcutaneously.

[0243] In certain embodiments, the method reduces the size of the cancer in question, for example, the size of the tumor. In some embodiments, the size of the cancer is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%.

[0244] In some embodiments, the method increases the overall survival of subjects. In some embodiments, overall survival increases compared to the average overall survival of subjects with the same cancer but treated with different treatments. In certain embodiments, overall survival increases by at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 2 times, at least about 3 times, or at least about 5 times. In some embodiments, overall survival increases by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 1 month, at least about 12 months, at least about 15 months, at least about 18 months, at least about 21 months, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years.

[0245] In some embodiments, the method increases the progression-free survival of subjects. In some embodiments, overall survival increases compared to the average progression-free survival of subjects with the same cancer but treated with different therapies. In certain embodiments, progression-free survival increases by at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 2 times, at least about 3 times, or at least about 5 times. In some embodiments, overall survival increases by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 1 month, at least about 12 months, at least about 15 months, at least about 18 months, at least about 21 months, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years.

[0246] In some embodiments, the method increases the objective response rate of the subject. In certain embodiments, the method induces a complete response in the subject. In some embodiments, the method induces a partial response in the subject.

[0247] In some embodiments, the method includes administering an anti-CD161 antibody (or polynucleotide, vector, or host cell) disclosed herein and performing a second therapy. In some embodiments, the second therapy is performed before administration of the anti-CD161 antibody. In some embodiments, the second therapy is performed after administration of the anti-CD161 antibody. In some embodiments, the second therapy is performed concurrently with administration of the anti-CD161 antibody. In certain embodiments, administration of the anti-CD161 antibody and performance of the second therapy are performed separately. In other embodiments, the anti-CD161 antibody and the second therapy are administered in a single formulation.

[0248] The second therapy can be any other therapy. In some embodiments, the second therapy includes immunotherapy. In some embodiments, the second therapy includes chemotherapy. In some embodiments, the second therapy includes radiation therapy. In some embodiments, the second therapy includes surgery. In some embodiments, the second therapy includes administering a second therapeutic agent.

[0249] In some embodiments, one or more additional therapeutic agents are a PD-1 antagonist, a TIM-3 inhibitor, a LAG-3 inhibitor, a TIGIT inhibitor, a CD112R inhibitor, a TAM inhibitor, a STING agonist, a 4-1BB agonist, or a combination thereof. In some embodiments, one or more additional therapeutic agents are a CD39 antagonist, a CD73 antagonist, a CCR8 antagonist, or a combination thereof. In some embodiments, the anti-CD73 is, for example, any anti-CD73 antibody disclosed in US Publication No. 2019 / 0031766A1, which is incorporated herein by reference in its entirety. In some embodiments, the anti-CD39 is, for example, any anti-CD39 antibody disclosed in International Publication No. WO2019 / 178269A2, which is incorporated herein by reference in its entirety.

[0250] In certain embodiments, the second therapeutic agent comprises a second antibody. In some embodiments, the second therapeutic agent comprises inducible T cell costimulator (ICOS), CD137 (4-1BB), CD134 (OX40), NKG2A, CD27, glucocorticoid-inducible TNFR-related protein (GITR), and herpesvirus entry mediator (HVEM), programmed death ligand-1 (PD-1), programmed death ligand-1 (PD-L1), CTLA-4, B lymphocyte and T lymphocyte attenuator (BTLA), T cell immunoglobulin and mucin domain-3 (TIM-3), lymphocyte activator gene-3 (LAG-3), and adenosyl The product contains an effective amount of antibodies that specifically bind to proteins selected from the following: A2a receptor (A2aR), killer cell lectin-like receptor G1 (KLRG-1), natural killer cell receptor 2B4 (CD244), CD160, T cell immune receptors including Ig and ITIM domains (TIGIT), T cell activation V domain Ig suppressor (VISTA), NKG2a, KIR, TGFβ, IL-10, IL-8, B7-H4, Fas ligand, CXCR4, mesothelin, CEACAM-1, CD96, CD52, HER2, and any combination thereof.

[0251] The anti-CD161 antibodies or their antigen-binding moieties described herein can replace or enhance previously or currently administered therapies. For example, during treatment with an anti-CD161 antibody or its antigen-binding moiety, the administration of one or more additional therapeutic agents can be discontinued or reduced, for example, by administering them at a lower dose. In some embodiments, the implementation of a previous therapy can be maintained. In some embodiments, the previous therapy is maintained until the level of the anti-CD161 antibody reaches a level sufficient to produce a therapeutic effect.

[0252] The anti-CD161 antibodies or their antigen-binding moieties described herein can be used as diagnostic agents. In some embodiments, the anti-CD161 antibody is used to identify subjects suitable for immunotherapy. In some embodiments, the anti-CD161 antibody is used to identify subjects having CD161 expression in NK cells.

[0253] In some embodiments, one or more additional therapeutic agents are PD-1 antagonists. In some embodiments, the PD-1 antagonists are selected from the group consisting of PDR001, nivolumab, pembrolizumab, pizilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, and AMP-224. In certain embodiments, one or more additional therapeutic agents are PD-L1 inhibitors. In some embodiments, the PD-L1 inhibitors are selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559. In some embodiments, the present disclosure provides a method for enhancing the activity of one or more anti-PD-1 antibodies (e.g., enhancing PD-1-mediated cytokine secretion from cells exposed to an anti-PD-1 antibody; enhancing anti-PD-1-mediated TNFα secretion; enhancing anti-PD-1-mediated IL-6 secretion), comprising exposing cells to the antibody or its antigen-binding portion provided by the present disclosure, either simultaneously with or subsequently to the anti-PD-1 antibody, thereby providing a method for enhancing the activity of one or more anti-PD-1 antibodies.

[0254] In some embodiments, one or more additional therapeutic agents include sunitinib (Sutent®), cabozantinib (CABOMETYX®), axitinib (INLYTA®), lenvatinib (LENVIMA®), everolimus (AFINITOR®), bevacizumab (AVASTIN®), epacadostat, and NKTR-214 (CD-122 biased type). Agonists), tivozanib (FOTIVDA®), abequinostat, ipilimumab (YERVOY®), tremelimumab, pazopanib (VOTRIENT®), sorafenib (NEXAVAR®), temsirolimus (TORISEL®), ramucirumab (CYRAMZA®), niraparib, savolitinib, bororanib (X-82), regorafe Nib (STIVARGO®), donafenib (multikinase inhibitor), camrelizumab (SHR-1210), pexastimodine debasilepvec (JX-594), ramucirumab (CYRAMZA®), apatinib (YN968D1), encapsulated doxorubicin (THERMODOX®), tivantinib (ARQ197), ADI-PEG20, binimetinib, apatinib These include silates, nintedanib, lirirumab, nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFIMZI®), semiprimab-rwlc (LIBTAYO®), tislerizumab, and / or spartalizumab.

[0255] In some embodiments, one or more additional therapeutic agents are TIM-3 inhibitors, which, by choice, are MGB453 or TSR-022.

[0256] In some embodiments, one or more additional therapeutic agents are LAG-3 inhibitors, which are optionally selected from the group consisting of LAG525, BMS-986016, and TSR-033.

[0257] In some embodiments, one or more additional therapeutic agents are TIGIT inhibitors. In some embodiments, one or more additional therapeutic agents are CD112R inhibitors. In some embodiments, one or more additional therapeutic agents are TAM (Axl, Mer, Tyro) inhibitors. In some embodiments, one or more additional therapeutic agents are STING agonists. In some embodiments, one or more additional therapeutic agents are 4-1BB agonists.

[0258] In some embodiments, one or more additional therapeutic agents are tyrosine kinase inhibitors, adenosine axis targeting agents (e.g., CD39 antagonists, CD73 antagonists or A2AR antagonists, A2BR antagonists or A2AR / A2BR dual antagonists), CCR8 antagonists, CTLA4 antagonists, VEG-F inhibitors, or combinations thereof.

[0259] III.A.1. Combination with chemotherapeutic agents Suitable chemotherapeutic agents for combination and / or co-administration with the compositions of the present invention include, for example, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracene, mitoxantrone, mitramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as their analogs or homologs. Further agents include, for example, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechloretamine, thioTEPA, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamine platinum(II) (DDP), procal Examples include vasine, altretamine, cisplatin, carboplatin, oxaliplatin, nedaplatin, satraplatin, or triplatin tetranitrate), anthracyclines (e.g., daunorubicin (formerly known as daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly known as actinomycin), bleomycin, mitramycin, and anthramycin (AMC)), and mitotic inhibitors (e.g., vincristine and vinblastine) and temozolomide.

[0260] III.A.2. Combination with PD-1 / PD-L1 Antagonists In some embodiments, the anti-CD161 antibody or its antigen-binding moiety provided by this disclosure is combined with one or more PD-1 antagonists that specifically bind to human PD-1 or PD-L1 and inhibit the biological activity and / or downstream pathway(s) and / or cellular processes mediated by human PD-1 / PD-L1 signaling or other human PD-1 / PD-L1-mediated functions (e.g., administered in combination).

[0261] Accordingly, the Specified provides PD-1 antagonists that directly or allosterically block, antagonize, suppress, inhibit or reduce PD-1 / PD-L1 biological activity, including downstream pathways and / or cellular processes mediated by PD-1 / PD-L1 signaling, such as receptor binding and / or induction of cellular responses to PD-1 / PD-L1. The Specified also provides PD-1 antagonists that reduce the quantity or amount of human PD-1 or PD-L1 produced by cells or subjects.

[0262] In some embodiments, the disclosure provides PD-1 antagonists that bind to human PD-1 and prevent, inhibit, or reduce the binding of PD-L1 to PD-1. In some embodiments, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and prevents translation. In some embodiments, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and causes degradation and / or turnover.

[0263] In some embodiments, PD-1 antagonists inhibit PD-1 signaling or function. In some embodiments, PD-1 antagonists block the binding of PD-1 to PD-L1, to PD-L2, or to both PD-L1 and PD-L2. In some embodiments, PD-1 antagonists block the binding of PD-1 to PD-L1. In some embodiments, PD-1 antagonists block the binding of PD-1 to PD-L2. In some embodiments, PD-1 antagonists block the binding of PD-1 to both PD-L1 and PD-L2. In some embodiments, PD-1 antagonists specifically bind to PD-1. In some embodiments, PD-1 antagonists specifically bind to PD-L1. In some embodiments, PD-1 antagonists specifically bind to PD-L2.

[0264] In some embodiments, a PD-1 antagonist inhibits the binding of PD-1 to its homologous ligand. In some embodiments, a PD-1 antagonist inhibits the binding of PD-1 to PD-L1, to PD-1 to PD-L2, or to both PD-L1 and PD-L2. In some embodiments, a PD-1 antagonist does not inhibit the binding of PD-1 to its homologous ligand.

[0265] In some embodiments, the PD-1 antagonist is an isolated antibody (mAb) or its antigen-binding fragment that specifically binds to PD-1 or PD-L1. In some embodiments, the PD-1 antagonist is an antibody or its antigen-binding fragment that specifically binds to human PD-1. In some embodiments, the PD-1 antagonist is an antibody or its antigen-binding fragment that specifically binds to human PD-L1. In some embodiments, the PD-1 antagonist is an antibody or its antigen-binding fragment that binds to human PD-L1 and inhibits the binding of PD-L1 to PD-1. In some embodiments, the PD-1 antagonist is an antibody or its antigen-binding fragment that binds to human PD-1 and inhibits the binding of PD-L1 to PD-1.

[0266] Several immune checkpoint antagonists that inhibit or disrupt the interaction between PD-1 and either or both of its ligands, PD-L1 and PD-L2, are in clinical development or are currently available to clinicians for the treatment of cancer.

[0267] Examples of anti-human PD-1 antibodies or antigen-binding fragments that may contain a PD-1 antagonist in any of the compositions, methods, and uses provided herein include KEYTRUDA® (pembrolizumab, MK-3475, h409A11; see US8952136, US8354509, US8900587, and EP2170959, all of which are referred to herein in whole). Intended by reference; Merck), OPDIVO(registered trademark) (nivolumab, BMS-936558, MDX-1106, ONO-4538; see US7595048, US8728474, US9073994, US9067999, EP1537878, US8008449, US8779105, and EP2161336, all of which are incorporated herein by reference in their entirety; Bristol Examples include, but are not limited to, Myers Squibb, MEDI0680 (AMP-514), BGB-A317 and BGB-108 (BeiGene), 244C8 and 388D4 (see WO2016106159, the whole of which is incorporated herein by reference; Enumeral Biomedical), PDR001 (Novartis), and REGN2810 (Regeneron). Thus, in some embodiments, the PD-1 antagonist is pembrolizumab. In some embodiments, the PD-1 antagonist is nivolumab.

[0268] Examples of anti-human PD-L1 antibodies or antigen-binding fragments that may contain a PD-1 antagonist in any of the compositions, methods, and uses provided herein include BAVENCIO® (avelumab, MSB0010718C, see WO2013 / 79174, the whole thereof incorporated herein by reference; Merck / Pfizer), IMFINZI® (durvalumab, MEDI4736), TECENTRIQ ( Examples include, but are not limited to, the registered trademark (atezolizumab, MPDL3280A, RG7446; see WO2010 / 077634, the whole thereof is incorporated herein by reference; Roche), MDX-1105 (BMS-936559, 12A4; see US7943743 and WO2013 / 173223, the whole thereof is incorporated herein by reference; Medarex / BMS), and FAZ053 (Novartis). Therefore, in some embodiments, the PD-1 antagonist is avelumab. In some embodiments, the PD-1 antagonist is durvalumab. In some embodiments, the PD-1 antagonist is atezolizumab.

[0269] In some embodiments, the PD-1 antagonist is an immunoadhesin that specifically binds to human PD-1 or human PD-L1, and is a fusion protein containing an extracellular or PD-1 binding moiety of PD-L1 or PD-L2 fused to a constant region, such as the Fc region of an immunoglobulin molecule. Examples of immunoadhesin molecules that specifically bind to PD-1 are described in WO2010 / 027827 and WO2011 / 066342, both of which are incorporated herein by reference in their entirety. In some embodiments, the PD-1 antagonist is AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein that specifically binds to human PD-1.

[0270] Those skilled in the art will understand that any PD-1 antagonist that binds to PD-1 or PD-L1 and disrupts the PD-1 / PD-L1 signaling pathway is suitable for the compositions, methods, and uses disclosed herein.

[0271] In some embodiments, PD-1 / PD-L1 antagonists are small molecules, nucleic acids, peptides, peptide mimes, proteins, carbohydrates, carbohydrate derivatives, or glycopolymers. Exemplary small molecule PD-1 inhibitors are described in Zhan et al., (2016) Drug Discov Today 21(6):1027-1036.

[0272] III.A.3. Combination with TIM-3 inhibitors In some embodiments, the anti-CD161 antibody or its antigen-binding moiety provided herein is combined with (e.g., administered in combination with) a TIM-3 inhibitor. The TIM-3 inhibitor may be an antibody, its antigen-binding fragment, an immunoadhesin, a fusion protein, or an oligopeptide. In some embodiments, the TIM-3 inhibitor is selected from MGB453 (Novartis), TSR-022 (Tesaro), or LY3321367 (Eli Lilly). In some embodiments, the anti-CD161 antibody or its antigen-binding moiety is administered in combination with MGB453. In some embodiments, the anti-CD161 antibody or its antigen-binding moiety is administered in combination with TSR-022.

[0273] III.A.4. Combination with LAG-3 inhibitors In some embodiments, the anti-CD161 antibody or its antigen-binding portion provided herein is combined with (e.g., administered in combination with) a LAG-3 inhibitor. The LAG-3 inhibitor may be an antibody, its antigen-binding fragment, an immunoadhesin, a fusion protein, or an oligopeptide. In some embodiments, the LAG-3 inhibitor is selected from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), TSR-033 (Tesaro), MK-4280 (Merck & Co), or REGN3767 (Regeneron).

[0274] III.A.5. Other Combinations In some embodiments, the anti-CD161 antibody or its antigen-binding moiety provided by this disclosure is combined with (e.g., administered in combination with) a TIGIT inhibitor, a kinase inhibitor (e.g., a tyrosine kinase inhibitor (TKI)), a CD112R inhibitor, a TAM receptor inhibitor, a STING agonist and / or a 4-1BB agonist, or a combination thereof. In some embodiments, the anti-CD161 antibody or its antigen-binding moiety provided by this disclosure is combined with (e.g., administered in combination with) a tyrosine kinase inhibitor, an adenosine axis-targeting agent (e.g., a CD39 antagonist, a CD73 antagonist or an A2AR antagonist, an A2BR antagonist or an A2AR / A2BR dual antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof.

[0275] III.B. Antibody Handling Procedures Anti-CD161 antibodies having VH and VL sequences disclosed herein can be used to create novel anti-CD161 antibodies by modifying the VH and / or VL sequences, or the constant region(s) bound to such sequences. Thus, in another embodiment described herein, the structural features of the anti-CD161 antibodies described herein can be used to create structurally related anti-CD161 antibodies that retain at least one functional property of the anti-CD161 antibodies described herein, such as inhibiting the interaction between human CD161 and CLEC2D. For example, one or more CDR regions of the antibodies disclosed herein can be recombinantly combined with known framework regions and / or other CDRs to create further recombinant anti-CD161 antibodies described herein, as described above. The starting material for the operation is one or more of the VH and / or VL sequences provided herein, or one or more CDR regions thereof. To create the manipulated antibody, it is not necessary to actually prepare an antibody having one or more of the VH sequences and / or VL sequences provided herein, or one or more of the CDR regions thereof (i.e., to express it as a protein). Rather, the information contained in the sequence(s) is used as a starting material to generate “second-generation” sequences(s) derived from the original sequence(s), and then the “second-generation” sequences(s) are prepared and expressed as proteins.

[0276] In addition, the antibodies disclosed herein may be improved by known techniques such as affinity maturation. Affinity maturation is a technique that allows for the selection of derived antibodies that bind to an antigen with higher affinity than the starting antibody. In some embodiments, the antibodies disclosed herein are used as starting antibodies for affinity maturation.

[0277] Therefore, this specification provides a method for preparing the anti-CD161 antibody described herein.

[0278] III.C. Antibody production The anti-CD161 antibodies described herein can be produced using a variety of known techniques, such as the standard somatic cell hybridization technique described in Kohler and Milstein, Nature 256:495 (1975). While somatic cell hybridization procedures are common, in principle, other techniques for producing monoclonal antibodies, such as viral or oncogenic transformation of B lymphocytes, or phage display techniques using libraries of human antibody genes, can also be employed.

[0279] In some embodiments, the animal system used to prepare hybridomas is the mouse system. Hybridoma production in mice is a very well-established procedure. Immunization protocols and techniques for isolating immunized splenocytes for fusion are known in the art. Fusion partners (e.g., mouse myeloma cells) and fusion procedures are also known.

[0280] Based on the sequence of the mouse monoclonal antibody prepared as described above, chimeric or humanized anti-CD161 antibodies can be prepared. DNA encoding heavy and light chain immunoglobulins can be obtained from the target mouse hybridoma and manipulated using standard molecular biological techniques to include non-mouse (e.g., human) immunoglobulin sequences. For example, to produce a chimeric antibody, the mouse variable region can be ligated to the human constant region using methods known in the art (see, for example, U.S. Patent No. 4,816,567 by Cabilly et al.). To produce a humanized antibody, the mouse CDR region can be inserted into the human framework using methods known in the art (see, for example, U.S. Patent No. 5,225,539 by Winter, and U.S. Patents No. 5,530,101, 5,585,089, 5,693,762 and 6,180,370 by Queen et al.).

[0281] In some embodiments, the anti-CD161 antibodies described herein are human monoclonal antibodies. Such human monoclonal antibodies against CD161 can be produced using transgenic or transchromosomal mice that possess a portion of the human immune system rather than a mouse lineage. These transgenic and transchromosomal mice include mice referred to herein as HuMAb mice and KM mice, respectively, and collectively referred to herein as "human Ig mice."

[0282] HUMAB-mouse (registered trademark) (Medarex, Inc.) contains miniloci of human immunoglobulin genes encoding unreconstituted human heavy chain (μ and γ) and κ light chain immunoglobulin sequences, along with targeted mutations that inactivate endogenous μ and κ chain loci (see, e.g., Lonberg, et al., (1994) Nature 368(6474):856-859). Therefore, these mice have reduced expression of mouse IgM or κ, and in response to immunization, the introduced human heavy and light chain transgenes undergo class switching and somatic mutation, generating high-affinity human IgGK monoclonals (Lonberg, N. et al. (1994) (see above); Lonberg, N. (1994) Handbook of Experimental Pharmacology 113:49-101; Lonberg, N. and Huszar, D. (1995) Intern. Rev. Immunol. 13:65-93, and Harding, F. and Lonberg, N. (1995) Ann. NYAcad. Sci. 764:536-546 review).

[0283] In some embodiments, the anti-CD161 antibodies described herein are produced using mice having a human immunoglobulin sequence on the transgene and transchromosome, for example, mice having a human heavy chain transgene and human light chain transchromosome. Such mice are referred to herein as “KM mice” and are described in detail in PCT Publication WO02 / 43478 by Ishida et al.

[0284] Furthermore, alternative transgenic animal lines expressing human immunoglobulin genes are also available and can be used to produce the anti-CD161 antibodies described herein. For example, an alternative transgenic line known as the Xeno mouse (Abgenix, Inc.) can be used, such as described, for example, in U.S. Patent No. 5,939,598.

[0285] Furthermore, alternative transchromosomal animal lines expressing human immunoglobulin genes are also available in the art and can be used to produce the anti-CD161 antibodies described herein. For example, mice possessing both human heavy chain transchromosomes and human light chain transchromosomes, known as "TC mice," can be used, and such mice are described in Tomizuka et al. (2000) Proc. Natl. Acad. Sci. USA 97:722-727. In addition, cattle possessing human heavy chain and light chain transchromosomes have been described in the art (Kuroiwa et al. (2002) Nature Biotechnology 20:889-894) and can be used to produce the anti-CD161 antibodies described herein.

[0286] Additional mouse lines described in the art for producing human antibodies, such as human anti-CD161 antibodies, include (i) Veloclmmune® mice (Regeneron Pharmaceuticals, Inc.) (in which case the endogenous mouse heavy and light chain variable regions are replaced by human heavy and light chain variable regions by homologous recombination and functionally linked to the endogenous mouse constant region; so that a chimeric antibody (human V / mouse C) can be produced in the mouse and subsequently converted to a full-length human antibody using standard recombinant DNA technology), and (ii) MeMo® mice (Merus Biopharmaceuticals, Inc.) (in which case the mouse contains a single reconstituted human common light chain variable region while containing an unreconstituted human heavy chain variable region). Such mice and methods for producing antibodies are described, for example, in US2012 / 0070861 and US2012 / 0073004.

[0287] The human monoclonal anti-CD161 antibodies described herein may also be prepared using phage display methods for screening libraries of human immunoglobulin genes. Such phage display methods for isolating human antibodies are well-established in the art. See, for example, U.S. Patents 5,223,409, 5,427,908, 5,969,108, and 5,885,793.

[0288] The human monoclonal anti-CD161 antibodies described herein can also be prepared using SCID mice in which human immune cells are reconstituted to produce a human antibody response upon immunization. Such mice are described, for example, in U.S. Patent No. 5,476,996.

[0289] Unless otherwise indicated, the implementation of this disclosure will employ conventional techniques in cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology within the scope of the art. Such techniques are adequately described in the literature. For example, Sambrook et al., ed. (1989) Molecular Cloning A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., ed. (1992) Molecular Cloning: A Laboratory Manual, (Cold Springs Harbor Laboratory, NY); DNGlover ed., (1985) DNA Cloning, Volumes I and II; Gait, ed. Synthesis; Mullis et al. U.S. Patent No. 4,683,195; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; Hames and Higgins, eds. (1984) Transcription And Translation; Freshney (1987) Culture Of Animal Cells (Alan R. Liss, Inc.); Immobilized Cells And Enzymes (IRL Press)(1986);Perbal (1984)A Practical Guide To Molecular Cloning;the treatise,Methods In Enzymology(Academic Press,Inc.,NY);Miller and Calos eds.(1987)Gene Transfer Vectors For Mammalian Cells, (Cold Spring Harbor Laboratory);Wu et al.,eds.,Methods In Enzymology,Vols.154 and 155;Mayer and Walker,eds.(1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London); Weir and Blackwell, eds., (1986) Handbook Of Experimental Immunology, Volumes I-IV; Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, (1986);); Crooks, Antisense drug Technology: Principles, strategies and applications,2. nd See Ed. CRC Press (2007) and Ausubel et al. (1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.).

[0290] The following examples are provided for illustrative purposes only and are not limiting. [Examples]

[0291] Example 1 Production of anti-CD161 monoclonal antibodies

[0292] Anti-CD161 monoclonal antibodies were generated using human Ig transgenic mice. A cohort of eight mice was immunized with human CD161-hFc protein (CD1-H5253) from AcroBiosystems, along with DNA immunogens expressing human and cyno CD161 ECD domains, including TCE-huCD161-ECD-67-225 and TCE-cyCD161-ECD-67-225. CD161-positive antibody titers were confirmed by flow cytometry on days 21 and 28 post-immunization using human and cyno CD161 overexpressing cell lines, including CHOK1:huCD161, CHOK1:cyCD161, and CHOK1:parental cells. The eight mice underwent four DNA immunizations and eight protein immunizations prior to the titer check on day 28. Plasma titer analysis on day 28 revealed strong CD161-positive antibody titers in all eight immunized mice.

[0293] Hybridoma generation and antibody screening assays

[0294] 32 days after immunization regimens using CD161 protein and DNA, spleens and lymph nodes were collected from eight immunized mice to generate hybridomas. Hybridoma fusion was performed with enriched B cells obtained from the spleens and lymph nodes, and the hybridomas were plated in 14 384-well plates. Eight days later, primary multiplex fluorescently labeled cell sorting (FACS) binding screening was performed, and CD161-positive binding hits were isolated using CHOK1:huCD161, CHOK1:cyCD161, and CHOK1:P cells (cell mixture). For hybridoma binding screening, 15 μL / well supernatant was transferred to a 384-well plate containing 15 μL of 20,000 / well cell mixture. The plates were incubated at 4°C for 30 minutes (min) and then washed 2.5 times with FACS buffer. Next, the cells were stained with 1 μg / mL AF647-goat α-human-Fc in 15 μL / well and incubated at 4°C for 30 minutes. Afterward, the cells were washed 2.5 times with FACS buffer and read using a flow cytometer. Anti-CD161 reference monoclonal antibody HP-3G10 (Biolegend; catalog number 39910) was used as a positive control.

[0295] Primary FACS-based screening identified 134 binders for human and cyno-CD161. As shown in Figure 3, of these hybridomas, those binding at a geometric mean fluorescence intensity (gMFI) greater than 50 K for HEK293T:huCD161 and a gMFI greater than 30 K for HEK293T:cyCD161 were selected for further validation. These CD161-positive binders were then evaluated for blocking or inhibiting the CD161-CLEC2D interaction. Of the 134 CD161-positive binders, 34 hybridomas that effectively blocked the interaction between CD161 and its ligand CLEC2D were selected for subcloning.

[0296] Screening assay: Cell-based binding assay using CD161 antibody

[0297] A cell-based binding assay using CD161 antibodies was performed to evaluate the cell surface binding ability of anti-CD161 antibodies against huCD161 antigen expressed on the surface of CHOK1 cells using FACS. Hybridoma antibody samples were screened using CHOK1 suspension cells expressing full-length membrane-bound huCD161 antigen. 50,000 cells were washed and transferred to a 96-well round-bottom plate, followed by the addition of 50 μl of purified antibody (0.001–10 μg / ml in Dulbecco's phosphate-buffered saline (DPBS)) and incubation at 4°C for 30 minutes. After incubation, the plate was centrifuged at 300 × g for 5 minutes. The cells were washed twice more with FACS buffer (1% FBS in DPBS) and stained with goat α-mouse IgG, Fcγ AF647 (1:2000 dilution). The plate was incubated in the dark at 4°C for 20 minutes. After incubation, the plates were centrifuged at 300×g for 5 minutes. For FACS analysis, the cells were washed twice more with FACS buffer and resuspended in 100 μl of FACS buffer. A commercially available anti-CD161 monoclonal antibody (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOCP1, BioXcell) was used as a negative control. Anti-CD161 antibodies generated from hybridomas can bind to the huCD161 antigen expressed on the surface of CHOK1 cells, as shown in Figures 5A-5D. Representative antibody clones, exemplified as 01D17, 03K18, 09H07, and HP-3G10 (control), were evaluated for binding to CHOK1 cells transfected with the full-length huCD161 construct, as shown in Figures 5A-5C. The overall dose-response curve is shown in Figure 5D.

[0298] Screening assay: ELISA-based binding assay using CD161 antibody

[0299] The binding interaction between purified anti-CD161 antibody from hybridoma supernatant and immobilized rhCD161-hFc was evaluated by ELISA. A high-binding-capacity 96-well ELISA plate was coated overnight at 4°C with 100 μl of DPBS containing 5 μg / mL of rhCD161-hFc protein (R&D). The plate was washed three times with ELISA wash buffer (Biolegend) and then blocked with 200 μl of DPBS containing 5% milk. The plate was then incubated at 37°C for 2 hours (hrs) on a gentle shaker at 120 rpm, and then washed three times with ELISA wash buffer. 100 μl of assay buffer (1% BSA in DPBS) containing 0.0001–10 μg / ml of purified antibody was added to each well, incubated at 37°C for 2 hours on a gentle shaker at 120 rpm, and then washed three times with ELISA wash buffer. Assay buffer containing goat anti-mouse detection antibody HRP (1:3000) was added at a rate of 100 μl / well and incubated at 37°C for 1 hour (h) on a gentle shaker at 120 rpm, followed by five washes with ELISA wash buffer. Then, 100 μl of TMB substrate was added per well and incubated at room temperature for 5 minutes. Subsequently, 100 μl of TMB stop solution was added to stop the reaction. The optical density (OD) of each well in the ELISA plate was read using a microplate reader set to a wavelength of 450 nm, as shown in Figure 4. A commercially available anti-CD161 monoclonal antibody (mAb) (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. As indicated by the positive OD values ​​shown in Figure 4, the anti-CD161 antibody can bind to immobilized rhCD161-hFc. Representative anti-CD161 antibody hybridoma clones, including 03K18, 06C21, 09H07, 12G06, and HP-3G10 (control), were evaluated by ELISA for their binding to rhCD161-coated plates.

[0300] Screening assays: Cell-based target and ligand blockade assays

[0301] The affinity of CLEC2D for CD161, as reported by Kamishikiryo, et al (2011), is K D The antibody concentration was 48 μM. The blockade of the CD161-CLEC2D interaction was evaluated using the antibody. The ability of anti-CD161-positive hybridoma hits to block the CD161-CLEC2D interaction was assessed using CHOK1 cells expressing huCLEC2D soluble protein and huCD161. Due to the low affinity between CD161 and CLEC2D, it was necessary to construct multimeric complexes of the CLEC2D protein to increase the valence.

[0302] To prepare CLEC2D polymers, biotinylated protein A (ThermoFisher) and hCLEC2D-hFc protein (Kactus) were mixed in a 1:20 ratio and incubated in the dark at 4°C for 5 minutes. PBS was added to the mixture and incubated in the dark at 4°C for 30 minutes. After incubation, streptavidin-APC beads (Biolegend) were added (diluted 1:1000 with DPBS) and incubated in the dark at 4°C for 30 minutes. Purified human IgG (ThermoFisher) was added to achieve Fc saturation.

[0303] Anti-CD161 antibody samples were screened using CHOK1 cells expressing the huCD161 antigen. 50,000–100,000 cells were washed and transferred to 96-well round-bottom plates. A pre-prepared hCLEC2D multimer complex (20 μl) + 50 μl of purified CD161 antibody sample was added to the CD161-expressing cells and incubated in the dark at 4°C for 30 minutes. After incubation, the plates were centrifuged at 300 × g for 5 minutes. For FACS analysis, cells were washed twice more with FACS buffer and resuspended in 100 μl of FACS buffer. A commercially available anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. The percentage of blockade of the CD161-CLEC2D interaction was calculated using the following formula: Blockade Rate (%) = (1 - CLEC2D multimers bound to CHOK1-CD161 in the presence of antibody % / CLEC2D multimers bound to CHOK1-CD161 in the absence of antibody %) × 100%. As shown in Figure 6, the hCLEC2D multimer effectively binds to CHOK1 expressing huCD161. A 10 ug / ml reference anti-CD161 monoclonal antibody (HP-3G10) resulted in complete blockade of the hCLEC2D multimer (Figure 6). Figure 7 shows the percentage of blockade by anti-CD161 antibodies that can block the interaction between the CLEC2D multimer and huCD161 expressed on the cell surface. Anti-CD161 antibodies obtained from selected hybridomas were evaluated by FACS and CLEC2D multimers bound to CD161-expressing CHOK1. Based on the blockage of the interaction between the hCLEC2D multimer and hCD161, the blockage percentages were ranked.

[0304] Screening assay: ELISA-based CD161:CLEC2D blockade assay

[0305] The ability of anti-CD161 antibodies to block the interaction between rhCLEC2D and immobilized rhCD161-hFc was evaluated by ELISA. A high-binding capacity 96-well ELISA plate was coated overnight at 4°C with 100 μl of DPBS containing rhCD161-hFc (5 μg / ml) protein (R&D). The plate was washed three times with ELISA wash buffer (Biolegend), then blocked with 200 μl of DPBS containing 5% milk, incubated at 37°C for 2 hours on a gentle shaker at 120 rpm, and then washed three times with ELISA wash buffer. 100 μl of assay buffer (1% BSA in DPBS) containing biotinylated 10 μg / ml CLEC2D-hFc (Kactus) and 0.03–66.7 nM purified antibody was added to each well, incubated at 37°C for 2 hours on a gentle shaker at 120 rpm, and then washed three times with ELISA wash buffer. 100 μl of assay buffer containing avidin HRP (1:1000) was added to each well of the plate and incubated at 37°C for 1 hour on a gentle shaker at 120 rpm, followed by five washes with ELISA wash buffer. 100 μl of TMB substrate was added to each well and incubated at 37°C for 30 minutes, then 100 μl of TMB stop solution was added to stop the reaction. The optical density (OD) of each well of the ELISA plate was read using a microplate reader set to a wavelength of 450 nm. A commercially available anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. As shown in Figure 8, the anti-CD161 antibody can block the interaction between rhCLEC2D and immobilized rhCD161-hFc. Representative anti-CD161 hybridoma clones, including 01D17, 03K18, 12G06, and HP-3G10 (positive control), were evaluated for competition with and disruption of interaction with the CLEC2D-CD161 contact site.

[0306] Example 2. Functional evaluation of anti-CD161 monoclonal antibody Evaluation of T cell activity induced by Staphylococcus enterotoxin B stimulation

[0307] Peripheral blood mononuclear cells (PBMCs) derived from healthy human donors were stimulated for 24 hours with 10 ng / ml Staphylococcus enterotoxin B (SEB) in complete cell culture medium (RPMI + 10% FBS) at 37°C, with or without anti-CD161 test antibody. Subsequently, IL-2 cytokine levels in the cell culture supernatant were measured. A commercially available anti-CD161 monoclonal antibody (mAb) (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. The percentage increase in IL-2 cytokine levels was calculated using the following formula: Percentage increase (%) = (Cytokine concentration in sample with antibody / Cytokine concentration in sample without antibody) × 100% - 100%.

[0308] Figure 9 shows anti-CD161 antibodies that can enhance T cell function by blocking the CD161-CLEC2D interaction, as measured by IL-2 cytokine release. This is a representative ELISA analysis of IL-2 cytokine expression in the supernatant of one donor's PBMCs. Anti-CD161 antibodies were ranked by the percentage increase in IL-2 based on IL-2 release levels compared to SEB-stimulated PBMCs without the test antibody (Figure 9).

[0309] T cell activity mediated by T cell engagers

[0310] IL-2 cytokine levels in PBMC supernatant were analyzed using an ELISA assay, and anti-CD161 antibodies were evaluated and ranked based on IL-2 release. In this assay, Raji cells were washed and transferred to 96-well round-bottom plates at 30,000 cells per well, and pre-incubated at 37°C for 30 minutes with 100 μg / ml CD3×CD19 T cell engager (InvivoGen). 90,000 Jurkat cells expressing full-length membrane-bound huCD161 antigen were added to the Raji / T cell engager plate with purified anti-CD161 antibody and human Fc blocker (1:1000). The plates were incubated at 37°C for 48 hours, and cytokine levels in the cell culture supernatant were subsequently measured. A commercially available anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control, and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. As shown in Figure 10, anti-CD161 antibodies can enhance Jurkat cells expressing huCD161 function, as measured by IL-2 release, by blocking the hCD161-hCLEC2D interaction.

[0311] Example 3. Anti-CD161 monoclonal antibody enhances T cell activity when inhibiting CD161-CLEC2D interaction. We assayed the ability of anti-CD161 monoclonal antibodies (mAbs) to rescue activation of MART-1 TCR+hCD161+ Jurkat cells by MeWo cells (HLA-A2+) overexpressing hCLEC2D in the presence of MART-1 peptide (HLA-A2 restricted).

[0312] method

[0313] MeWo cell lines (HLA; A*0201, required for peptide presentation) overexpressing Mock-GFP and hCLEC2D-GFP were cultured overnight in FB96-well plates at 20K / well (see Figure 11). The following day, the plates were centrifuged at 300g / 5min, the culture medium was carefully removed without touching the adhered MeWo cells, and then 0.1ug / well of MART-1 peptide prepared in RPMI complete medium was added, and the cells were incubated at 37°C for 30 minutes. During incubation, round-bottom plates of 50μL of Jurkat-MART-1 TCR cells overexpressing human CD161 (100K / well) were prepared, and then 50μL of anti-CD161 mAb was added at a final concentration of 0.005-10μg / mL. After incubation of the peptide plate, Jurkat-MART1-CD161 OE cell line containing 100 μL of anti-CD161 mAb was added to MeWo cells containing peptide FB in a 96-well plate and incubated at 37°C for 24 hours. The following day, after incubation, the plate was incubated at 500 g for 5 minutes, 180 μL of supernatant was collected, and IL-2 ELISA was performed using the Biolegend kit (catalog number 431816, lot: B352552; follow manufacturer's instructions).

[0314] result

[0315] Each anti-CD161 mAb induced a dose-dependent increase in TCR activation compared to the isotype control, as measured by IL-2 secretion (Figure 12). These results indicate that TCR activation is influenced by human CLEC2D, and this can be rescued by anti-CD161 antibodies that block the interaction between the CD161 receptor (Jurkat) and its ligand CLEC2D expressed on MeWo cells. As expected, the control, the hIgG1LALA-PG antibody, did not further increase TCR activation.

[0316] Therefore, anti-CD161 antibodies inhibit the negative interaction between CD161 and CLEC2D, enhancing T cell activation as measured by IL-2 release.

[0317] Example 4. Anti-CD161 monoclonal antibody binds to TALL-104 cells and rescues TALL-104 cells from cytolysis of hCLEC2D overexpressing target cells. Anti-CD161 mAbs were assayed for their ability to bind to TALL-104 effector cells and to rescue hCLEC2D overexpressing (OE) PC3 target cells from cytolysis by CD161+ TALL-104 (CD3 / TCR+ CD4- CD8+ CD56+ CD16-) effector cells. TALL-104 effector cells are a clinically relevant MHC-unrestricted human cytotoxic T cell line.

[0318] TALL-104 combination

[0319] TALL-104 cells were washed, counted, and then plated at 100,000 cells / well in a 96-well round-bottom plate. The plate was centrifuged at 500g for 5 minutes, and the supernatant was discarded. Next, 50 μL of anti-CD161 mAb prepared in FACS buffer was added, the sample was mixed, and then incubated on ice for 30 minutes. After incubation, the plate was washed with 100 μL of FACS buffer (DPBS + 1% FBS), centrifuged at 500g for 5 minutes, and the supernatant was discarded. Next, the plate was incubated on ice for 20 minutes with 50 μL of goat anti-human IgG(H+L) cross-adsorbed secondary antibody, ALEXA FLUOR® Plus 647 and zombie aqua, and live-dead dyes diluted 1:2000. After incubation, the plate was washed with 100 μL of FACS buffer (DPBS + 1% FBS). Next, the plate was centrifuged at 500g for 5 minutes. Then, the supernatant was discarded, and the cell pellet was resuspended in 100μL / well of FACS buffer and acquired using a flow cytometer.

[0320] Each of the anti-CD161 mAbs tested was able to bind to TALL-104 cells in a dose-dependent manner (Figure 13).

[0321] Rescue of TALL-104 effect pedal function

[0322] Using an xCELLigence-based cytotoxicity assay, we evaluated the ability of anti-CD161 mAbs to rescue hCLEC2D-overexpressing PC3 target cells from cytolysis by CD161+ TALL-104 effector cells (CD3 / TCR+ CD4- CD8+ CD56+ CD16-) (Figure 14A). CD161+ TALL-104 human killer CD8+ T cells were incubated with CLEC2D+ tumor cells (PC3-CLEC2D OE). Overexpression of CLEC2D in PC3 tumor cells inhibited TALL-104-mediated killing compared to WT PC3 cells (CLEC2D-negative) (Figures 14B-14C). Anti-CD161 mAbs, which were shown to inhibit the CD161-CLEC2D interaction above, were tested for their ability to enhance TALL-104-mediated killing against PC3-CLEC2D OE.

[0323] Step 1: Adhesion target cells (i.e., tumor cells - PC3)

[0324] -On day 1, Mock / OEPC-3 cells were trypsin-treated and counted. The cells were then plated overnight in 96-well E-plates at 10K cells / well.

[0325] Step 2: Addition of TALL-104 as effector cells (i.e., immune cells)

[0326] The following day, TALL-104 cells were counted and centrifuged. The TALL-104 cells were then resuspended in culture medium and treated with anti-CD161 mAb or isotype IgG1 LALA-PG control for 30 minutes at 37°C. After incubation, TALL-104 cells were placed in an E-plate with PC3 cells in a 1:1 ratio and incubated at room temperature for 30 minutes to promote uniform distribution of TALL-104 cells over the PC3 target cells. The plate was then returned to the xCELLigence instrument placed in the incubator, and data were acquired at 37°C for 20 hours.

[0327] Step 3: If effector cells induce the destruction of target adherent tumor cells, this cytolytic activity can be detected with high sensitivity and accuracy.

[0328] The xCELLigence RTCA label-free technology counts cells using impedance changes of gold electrodes embedded in a dedicated E-plate. In this xCELLigence assay, hCLEC2D OE PC3 tumor cells adhere to the surface of staggered gold microelectrodes embedded at the bottom of a microtiter plate (E-plate). When target cells interact with the gold sensors, an impedance signal is generated that reflects the number, size, and adhesion strength of the target cells. The fact that the effector TALL-104 cells are not inherently adherent simplifies the system, as these cells generate a low impedance signal that can be easily subtracted. On the other hand, when TALL-104 attacks and kills PC3 cells by cytolysis, the viability of the target PC3 cells is reflected in real-time impedance changes, providing a kinetic assessment of the killing. The cytolytic activity of TALL-104 causes adherent cells to round up and detach, resulting in a decrease in the CI value.

[0329] The RTCA system uses cell impedance readout to monitor real-time changes in cell number, cell size, and cell substrate adhesion strength as a single parameter called the cell index (CI), which reflects the viability of target PC3 cells. We demonstrate the effectiveness of this approach as a efficacy assay for TALL-104 cell-mediated cytolysis of target PC3 cells. First, PC3 prostate cancer cells were manipulated to either overexpress a mock (Mock-PC3) or overexpress human CLEC2D (CLEC2D OE PC3) as controls, cultured in E-plates, and their growth was measured using xCELLigence. The day after seeding PC3 target cells, TALL-104 effector cells were added to the wells in a 1:1 E:T ratio, and the viability of target PC3 cells was monitored. PC3 cells treated only with effector cell growth medium served as a negative control. Addition of TALL-104 cells to PC3 cells resulted in an immediate and time-dependent decrease in CI. The PC3 signal decreased approximately 24 hours after the addition of effector cells due to cell lysis.

[0330] Each anti-CD161 mAb showed dose-dependent increased cytolysis only in human CLEC2D OE-PC3 cells (Figure 15). This indicates that TALL-104-based PC3 killing is affected by human CLEC2D and can be rescued by anti-CD161 antibodies that block the interaction between the CD161 receptor (TALL-104) and its ligand CLEC2D expressed on PC3 cells.

[0331] Example 5. Anti-CD161 mAb increases NK cell toxicity. The anti-CD161 mAb was assayed for its effect on the cytotoxicity of primary human NK cells. Human NK cells were activated overnight with 200 U / mL of IL-2 and then co-cultured with CellTrace Violet (CTV)-labeled PC-3 CLEC2D-OE cells (CLEC2D overexpression) or Raji cells for 18 hours in the presence of either Fc block + 1 μg / mL of anti-CD161 mAb or hIgG1-LALAPG control antibody. Cytotoxicity against tumor cells was measured by the survival rate of tumor cells (Zombie+) within the CTV+ population.

[0332] Upon contact with anti-CD161 mAb, when evaluated by relative tumor killing ability, NK cell cytotoxicity against PC-3 CLEC2D-OE cells (Figure 16A) or Raji cells (Figure 16B) increased compared to the isotype control. Relative tumor killing ability was calculated by normalizing the percentage of tumor cell death in the presence of anti-CD161 mAb to the isotype. The data represent 9 healthy donors for PC-3 CLEC2D-OE (Figure 16A) and 7 healthy donors for Raji (Figure 16B) in 4 or 3 independent assays for each anti-CD161 clone.

[0333] Example 6. Anti-CD161 mAb enhances tumor growth inhibition by NK cells Healthy naive female mice (6 - 8 weeks old; NOD.Cg-Prkdc scid Il2rg tm1Wjl / SzJ NSG; Jackson Laboratory) were subcutaneously injected with 2 × 10 6 individual PC3-CLEC2D-OE cells. When the tumor size reached approximately 70 mm 3 (day 4), the mice were randomized into treatment groups (n = 4). The first 2 doses (10 mg / kg) of anti-CD161 antibody 12G06 were injected intravenously, followed by 6 doses (15 mg / kg) injected intraperitoneally. Treatment was initiated on day 4 and repeated every 3 days.

[0334] All mice received 5 × 10 6Two intratumoral administrations of individual human NK cells were given. NK cells were freshly isolated from cryopreserved human PBMCs and cultured in NK MACS medium (Miltenyi Biotech) with recombinant human IL-2 at 100 U / mL on the night before injection.

[0335] Tumor growth was measured two-dimensionally with calipers, and tumor volume (mm 3 ) was calculated using the formula (short diameter 2 × long diameter) / 2.

[0336] Administration of the 12G06 antibody significantly enhanced tumor growth inhibition in the presence of NK cells compared to the vehicle control group in the xenograft model (Figure 17). These experiments are repeated with additional anti-CD161 antibodies (e.g., 01C07, 09O13, 12H24, 06M03S, and / or 06M03L).

[0337] Example 7. Anti-CD161 mAb enhances tumor growth inhibition by co-transplanted PBMCs Healthy mice (NOD.Cg-Prkdc scid Il2rg tm1Wjl / SzJ NSG; Jackson Laboratory) were subcutaneously injected on the right flank with a 1:4 mixture of PBMCs and U-87MG (glioblastoma) human tumor cells at 4.5 × 10 6 cells. One hour after transplantation, the anti-CD161 antibody 12G06 was administered intraperitoneally. The mice received a total of 8 doses of 12G06 at 20 mg / kg every 3 days.

[0338] Tumor growth was measured two-dimensionally with calipers, and tumor volume (mm 3 ) was calculated using the formula (short diameter 2 × long diameter) / 2.

[0339] Administration of the 12G06 antibody significantly enhanced tumor growth inhibition in the presence of human PBMCs compared to the vehicle control group in the xenograft model (Figure 18). These experiments are repeated with additional anti-CD161 antibodies (e.g., 01C07, 09O13, 12H24, 06M03S, and / or 06M03L).

[0340] Example 8. In vivo (Prophetic) analysis of anti-CD161 antibodies in human cancer patients. The safety and efficacy of anti-CD161 antibodies will be tested in human cancer patients. Patients will receive 12G06, 01C07, 09O13, 12H24, 06M03 S, and / or 06M03 L antibodies. Patients will be monitored for all adverse events, and tumor tissue burden, progression-free survival, and overall survival will be determined.

[0341] It should be understood that the section describing embodiments for carrying out the invention, rather than the section describing the summary and abstract of the invention, is intended to be used to interpret the claims. The summary and abstract section may describe one or more, but not all, exemplary embodiments of the disclosure intended by the inventor(s) and is therefore not intended to limit the scope of the disclosure and the appended claims in any way.

[0342] This disclosure is described above using functional units that demonstrate the implementation of specific functions and their relationships. The boundaries of these functional foundational elements are arbitrarily defined herein for the sake of explanation. Different boundaries may be defined, insofar as the specific functions and their relationships are adequately implemented.

[0343] The foregoing descriptions of specific embodiments are sufficient to illustrate the general nature of the disclosure and can be easily modified and / or adapted for various uses by applying the knowledge of a person skilled in the art without excessive experimentation and without departing from the general concepts of the disclosure. Such adaptations and modifications are therefore intended to be within the meaning and scope of equivalent embodiments of the embodiments of the disclosure, based on the teachings and guidance provided herein. It should be understood that the expressions and terms used herein are for illustrative purposes only, not limiting purposes, and that they are to be interpreted by a person skilled in the art in light of these teachings and guidance.

[0344] The scope and breadth of this disclosure should not be limited by any of the exemplary embodiments described above, but should be defined solely by the following claims and their equivalents.

[0345] All cited references throughout this application (including references, U.S. or foreign patents or patent applications, and websites) are expressly incorporated herein by reference as if they were included in their entirety for any purpose. In the event of any conflict, the content disclosed herein in writing shall prevail.

Claims

1. An antibody or its antigen-binding portion that specifically binds to CD161, comprising a heavy chain variable region (VH) and a light chain variable region (VL), The aforementioned VH includes VH complementarity determination region 1 (VH-CDR1), VH-CDR2, and VH-CDR3. The aforementioned VL includes VL-CDR1, VL-CDR2, and VL-CDR3. The VH-CDR3 contains an amino acid sequence selected from the sequences described in Sequence IDs 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323, and 333. The antibody or its antigen-binding portion.

2. The antibody or antigen-binding moiety according to claim 1, wherein the VH-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, and 332.

3. The antibody or antigen-binding moiety according to claim 1 or 2, wherein the VH-CDR1 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, and 331.

4. The antibody or antigen-binding moiety according to any one of claims 1 to 3, wherein the VL-CDR3 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, and 336.

5. The antibody or antigen-binding moiety according to any one of claims 1 to 4, wherein the VL-CDR2 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, and 335.

6. The antibody or antigen-binding moiety according to any one of claims 1 to 5, wherein the VL-CDR1 comprises an amino acid sequence selected from the sequences described in SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, and 334.

7. (i) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (ii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 91, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 92, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 93, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 94, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 95, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 96; (iii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 111, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 112, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 113, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 114, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 115, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 116; (iv) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 141, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 142, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 143, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 144, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 145, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 146; (v) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 211, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 212, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 213, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 214, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 215, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 216; (vi) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 221, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 222, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 223, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 224, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 225, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 226; (vii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 271, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 272, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 273, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 274, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 275, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 276; (viiii) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 281, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 282, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 283, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 284, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 285, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 286; (ix) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 291, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 292, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 293, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 294, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 295, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 296; (x) VH-CDR1 containing the amino acid sequence described in SEQ ID NO: 301, VH-CDR2 containing the amino acid sequence described in SEQ ID NO: 302, VH-CDR3 containing the amino acid sequence described in SEQ ID NO: 303, VL-CDR1 containing the amino acid sequence described in SEQ ID NO: 304, VL-CDR2 containing the amino acid sequence described in SEQ ID NO: 305, and VL-CDR3 containing the amino acid sequence described in SEQ ID NO: 306; or Any combination of (xi)(i) to (x) The antibody or its antigen-binding portion according to any one of claims 1 to 6, comprising:

8. The antibody or antigen-binding moiety according to any one of claims 1 to 7, wherein the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337.

9. The antibody or antigen-binding moiety according to any one of claims 1 to 8, wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

10. An antibody or its antigen-binding moiety that specifically binds to CD161, comprising a variable heavy chain (VH) domain and a variable light chain (VL) domain, wherein the VH is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 98% of an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. The antibody or its antigen-binding moiety, comprising an amino acid sequence having 99% sequence identity, wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

11. The antibody or antigen-binding moiety according to claim 1 to 10-1, wherein the VH comprises an amino acid sequence selected from SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337.

12. The antibody or antigen-binding moiety according to any one of claims 1 to 11, wherein the VL comprises an amino acid sequence selected from SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

13. (i) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 7, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 8; (ii) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 97, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the amino acid sequence selected from SEQ ID NO: 98; (iii) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 117, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 118; (iv) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 147, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 148; (v) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 217, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 218; (vi) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 227, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 228; (vii) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 277, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 278; (viiii) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 287, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 288; (ix) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 297, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 298; (x) The VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 307, and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 308; or Any combination of (xi)(i) to (x), The antibody or its antigen-binding portion according to any one of claims 1 to 12.

14. (i) VH containing the amino acid sequence described in SEQ ID NO: 7 and VL containing the amino acid sequence described in SEQ ID NO: 8; (ii) VH containing the amino acid sequence described in SEQ ID NO: 97 and VL containing the amino acid sequence described in SEQ ID NO: 98; (iii) VH containing the amino acid sequence described in SEQ ID NO: 117 and VL containing the amino acid sequence described in SEQ ID NO: 118; (iv) VH containing the amino acid sequence described in SEQ ID NO: 147 and VL containing the amino acid sequence described in SEQ ID NO: 148; (v) VH containing the amino acid sequence described in SEQ ID NO: 217 and VL containing the amino acid sequence described in SEQ ID NO: 218; (vi) VH containing the amino acid sequence described in SEQ ID NO: 227 and VL containing the amino acid sequence described in SEQ ID NO: 228; (vii) VH containing the amino acid sequence described in SEQ ID NO: 277 and VL containing the amino acid sequence described in SEQ ID NO: 278; (viiii) VH containing the amino acid sequence described in SEQ ID NO: 287 and VL containing the amino acid sequence described in SEQ ID NO: 288; (ix) VH containing the amino acid sequence described in SEQ ID NO: 297 and VL containing the amino acid sequence described in SEQ ID NO: 298; (x) VH containing the amino acid sequence described in SEQ ID NO: 307 and VL containing the amino acid sequence described in SEQ ID NO: 308; Any combination of (xi)(i) to (x) The antibody or its antigen-binding portion according to any one of claims 1 to 13, including

15. An antibody or its antigen-binding portion that binds to the same epitope as the antibody or its antigen-binding portion described in any one of claims 1 to 14.

16. An antibody or its antigen-binding portion that cross-competes with the antibody or its antigen-binding portion according to any one of claims 1 to 14 in terms of binding to CD161.

17. K is less than approximately 1000 nM, less than approximately 500 nM, less than approximately 100 nM, less than approximately 50 nM, or less than approximately 10 nM. Ｄ The antibody or its antigen-binding portion according to any one of claims 1 to 16, which binds to CD161.

18. K below approximately 50 nM Ｄ The antibody or its antigen-binding portion according to any one of claims 1 to 17, which binds to CD161.

19. K less than approximately 10 nM Ｄ The antibody or its antigen-binding portion according to any one of claims 1 to 18, which binds to CD161.

20. An antibody or its antigen-binding moiety according to any one of claims 1 to 19, which inhibits the interaction between CD161 and member D of the C-type lectin domain family 2 (CLEC2D).

21. An antibody or its antigen-binding moiety according to any one of claims 1 to 20, which is capable of inducing or enhancing the production of one or more cytokines by immune cells.

22. The antibody or antigen-binding moiety according to claim 21, wherein the one or more cytokines include IL2, TNFa, IFNg, or any combination thereof.

23. The antibody or its antigen-binding portion according to any one of claims 1 to 22, wherein the antigen-binding portion of the antibody comprises VHH, vNAR, microbody, nanobody, scFv, or any combination thereof.

24. A multispecific antibody comprising the antibody or its antigen-binding moiety according to any one of claims 1 to 23.

25. A bispecific antibody comprising the antibody or its antigen-binding moiety according to any one of claims 1 to 23.

26. A nucleic acid molecule or set of nucleic acid molecules encoding an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, or a bispecific antibody according to claim 25.

27. A vector or set of vectors comprising a nucleic acid molecule or a set of nucleic acid molecules as described in claim 26.

28. A viral vector, the vector or set of vectors according to claim 36.

29. A host cell comprising a nucleic acid molecule or set of nucleic acid molecules as described in claim 26, or a vector or set of vectors as described in claim 27 or 28.

30. A pharmaceutical composition comprising an antibody or antigen-binding moiety thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, or a host cell according to claim 29, and a pharmaceutically acceptable carrier.

31. A method for treating a disease or disorder in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

32. The method according to claim 31, wherein the disease or disorder includes cancer.

33. A method for inducing an immune response in a subject requiring such response, comprising administering to the subject an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

34. The method according to claim 33, wherein the subject is suffering from cancer.

35. A method for treating cancer in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

36. The aforementioned cancers include acoustic neuroma, acute lymphoblastic leukemia, acute myeloid leukemia, adenocarcinoma, and cancers and carcinomas of the urinary tract, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, biliary tract cancer, bladder cancer, bone cancer, brain cancer, brainstem glioma, breast cancer, bronchogenic carcinoma, Burkitt lymphoma and marginal zone B-cell lymphoma, adrenal cancer, anal cancer, gastrointestinal cancer, endocrine cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical carcinoma, endometrial carcinoma, fallopian tube carcinoma, renal pelvis carcinoma, vaginal carcinoma, vulvar carcinoma, and central nervous system (CNS) Cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, craniopharyngioma, cystadenocarcinoma, fetal cancer, endometrial cancer, endosarcoma, environmentally induced cancer (including those induced by asbestos), ependymoma, epidermal carcinoma, epithelial carcinoma, esophageal cancer, esophageal carcinoma, Ewing's tumor, eye cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, germ cell tumor, glioblastoma (e.g., glioblastoma multiforme), glioma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatoma, Hodgkin's disease, carcinoma in situ, Kaposi's sarcoma, kidney cancer (e.g., renal cell carcinoma) Cellular carcinoma (RCC), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung carcinoma, intralymphatic sarcoma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyeloblastic myelomonocytic monocytic erythroleukemia, myxosarcoma, nasopharyngeal cancer, neoplasms of the central nervous system (CNS), neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer (NSCLC), non-small cell lung carcinoma, oligodendroglioma, oral cancer (e.g., lips, tongue, mouth and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary carcinoma, papillary carcinoma, pediatric sarcoma, pineal glandoma, Pituitary adenoma, polycythemia vera lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate cancer), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, soft tissue sarcoma, sebaceous gland carcinoma, seminomas, nasal cavity / paranasal sinus natural killer cancers, skin cancer, small cell lung cancer (SCLC), pediatric solid tumors, spinal axial tumors, squamous cell carcinoma, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synoviomas, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-associated cancer or cancer of viral origin (e.g., human papillomavirus (HPV-associated or HPV-derived tumors)),The method according to any one of claims 32, 34, and 35, selected from Waldenström macroglobulinemia and Wilms' tumor; and any combination of the aforementioned cancers.

37. A method for treating an infectious disease in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

38. The aforementioned infectious disease, (i) Infections caused by influenza, herpes, giardia, malaria, leishmania, or any combination thereof; (ii) Infections caused by human immunodeficiency virus (HIV), hepatitis virus, herpesvirus, adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus, or arbovirus encephalitis virus, or any combination thereof; (iii) Infections caused by Chlamydia, Rickettsia, Mycobacteria, Staphylococcus, Streptococcus, Pneumococcus, Meningococcus, Conococcus, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacillus, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease, or any combination thereof; (iv) Infections caused by Candida, Cryptococcus neoformans, Aspergillus, Mucorales, Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, or Histoplasma capsulatum, or any combination thereof; (v) Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp. , Giardia lambia, Cryptosporidium sp. Infections caused by Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, or Nippostlongylus brasiliensis, or any combination thereof; or (vi) Any combination of (i) to (v) The method according to claim 37, including the method described in claim 37.

39. A method for treating an autoimmune disease in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

40. A method for activating immune cells, comprising contacting the immune cells with an antibody or antigen-binding portion thereof according to any one of claims 1 to 23, a multispecific antibody according to claim 24, a bispecific antibody according to claim 25, a nucleic acid molecule or set of nucleic acid molecules according to claim 26, a vector or set of vectors according to claim 27 or 28, a host cell according to claim 29, or a pharmaceutical composition according to claim 30.

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