Anti-IL2 receptor gamma antigen binding protein

Abstract

Antibodies that bind to IL2 receptor gamma proteins and methods of using the same, e.g., to treat or prevent disease, are provided.SOLUTION: The present invention provides antibodies and antigen-binding fragments (e.g., human antibodies) that specifically bind to human IL2 receptor gamma (IL2R γ). Methods of using the antibodies and fragments to treat or prevent diseases mediated by IL2R γ (e.g., graft versus host disease) are also provided, along with methods of making the antibodies and fragments.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application claims the benefits of U.S. Provisional Patent Application No. 62 / 799,851, filed on February 1, 2019, and the entire Provisional Patent Application is incorporated herein by reference.

[0002] Field of Invention The present invention relates to an antibody that binds to the anti-IL2 receptor gamma protein, and to a method of using the same for, for example, treating or preventing a disease. [Background technology]

[0003] The common cytokine receptor gamma chain (γc) was first identified as the third chain of the interleukin-2 (IL-2) receptor complex and named IL-2Rγ. The same subunit has been identified as a part of several other cytokine receptor complexes: IL-4, IL-7, IL-9, IL-15, and IL-21, and is therefore sometimes referred to as γc (common cytokine receptor gamma chain). In addition to signaling of these cytokine receptors, γc is involved in ligand binding.

[0004] The binding of cytokines to their receptors activates the Janus kinase (JAK) family proteins, tyrosine kinases JAK1 and JAK3, triggering transphosphorylation of JAK1 and JAK3 on the tyrosine. JAK1 associates with JAK3 and its unique α or β chain using the receptor's γc. Phosphorylated JAK can then activate signal transducer and activator of transcription (STAT) proteins, which together form the JAK / STAT signaling pathway. Phosphorylation of STAT leads to dimerization, thereby acquiring high-affinity DNA-binding activity and translocating to the nucleus. There, they act as transcription factors that induce transcription of target genes.

[0005] The γc gene (IL2RG) is located on chromosome Xq13. IL-2Rγ is mutated in patients with X-linked severe combined immunodeficiency (X-SCID). Patients with this disease exhibit severe immunodeficiency due to a lack of T, NK, and fully mature B cells.

[0006] IL-7, -9, and -15 are associated with psoriasis and rheumatoid arthritis (Non-patent documents 1-6).

[0007] Blocking IL-4 and IL-9 has been shown to improve asthma symptoms in mice (Non-Patent Documents 7-11).

[0008] IL-21 has been linked to various inflammatory disorders, including Crohn's disease and rheumatoid arthritis. (Non-patent literature 12-13). [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Pathak, “The expanding role of IL-7 and thymic stromal lymphopoietin as therapeutic target for rheumatoid arthritis.” Expert Opin Ther Targets.18(5):581–94 (2014) [Non-Patent Document 2] Hughes-Austin et al., “Multiple cytokines and chemokines are associated with rheumatoid arthritis-related autoimmunity in first-degree relatives without rheumatoid arthritis:Studies of the Aetiology of Rheumatoid Arthritis (SERA),” Ann Rheum Dis.;72(6):901–7 (2013) [Non-Patent Document 3] Dantas et al., “Increased Serum Interleukin-9 Levels in Rheumatoid Arthritis and Systemic Lupus Erythematosus: Pathogenic Role or Just an Epiphenomenon?”, Dis Markers.2015;2015:519638 [Non-Patent Document 4] Yang et al., “Therapeutic potential of IL-15 in rheumatoid arthritis,” Hum Immunol. November 2015; 76(11): 812–8. [Non-Patent Document 5] Lesiak et al., “Are interleukin-15 and -22 a new pathogenic factor in pustular palmoplantar psoriasis?”, Postepy Dermatol Alergol.33(5):336–339 ​​(2016) [Non-Patent Document 6] Raeber et al., “The role of cytokines in T-cell memory in health and disease,” Immunol Rev. 283(1): pp. 176–193 (2018) [Non-Patent Document 7] Generoso et al., “Prospects for Monoclonal Antibody Therapy in Pediatric Asthma,” Curr Allergy Asthma Rep. 18(9):45 (2018) [Non-Patent Document 8] Tashkin & Wechsler, “Role of eosinophils in airway inflammation of chronic obstructive pulmonary disease”, Int J Chron Obstruct Pulmon Dis.13:335-349 (2018) [Non-Patent Document 9] Buzney et al., “Asthma and Atopic Dermatitis: A Review of Targeted Inhibition of Interleukin-4 and Interleukin-13 As Therapy for Atopic Disease,” J Drugs Dermatol. 15(2): pp. 165–71 (2016) [Non-Patent Document 10] Lloyd & Harker, "Epigenetic Control of Interleukin-9 in Asthma", N Engl J Med.379(1):87-89 (2018) [Non-Patent Document 11] Neurath & Finotto, "IL-9 signaling as key driver of chronic inflammation in mucosal immunity", Cytokine Growth Factor Rev. 29:93-9 (2016) [Non-Patent Document 12] Holm et al., "Evaluating IL-21 as a Potential Therapeutic Target in Crohn's Disease," Gastroenterol Res Pract.2018:5962624 (2018) [Non-Patent Document 13] Dinesh & Rasool, “Multifaceted role of IL-21 in rheumatoid arthritis: Current understanding and future perspectives”, J Cell Physiol.233(5):3918~3928 (2018) [Overview of the project] [Means for solving the problem]

[0010] The present invention relates to approximately 2.75 × 10 at 25°C. -9 M ~ approx. 3.36×10 -7 M's KD Binding to human IL2Rγ at; about 6.42×10 -9 M to about 3.53×10 -7 M of K D Binding to human IL2Rγ at; or about 3.53×10 -7 M lower K D Binding at; about 3.18×10 -9 M to about 2.38×10 -7 M of K D Binding to cynomolgus monkey (Macaca fascicularis) IL-2Rγ at; about 8.29×10 -9 M to about 3.20×10 -7 M of K D Binding to cynomolgus monkey IL-2Rγ at; or about 3.20×10 -7 M lower K D Binding at; about 2.45×10 -9 M to about 1.20×10 -8 M of K D Binding to human IL2Rγ at; or about 1.20×10 -8 M lower K D Binding at; about 1.86×10 -11 M to about 3.00×10 -8 M of K D Binding to human IL2Rγ at; or about 3.00×10 -8 M lower K D Binding at; about 1.84×10 -8 M, 3.76×10 -9 M, 1.08×10 -7 M, 2.17×10 -8 M, 6.02×10 -9 M or 7.93×10 -8 M of K D Binding to mouse IL2Rγ at; or not detectably binding; about 5.59×10 -8 M, 6.11×10 -9 M, 3.87×10 -7 M, 5.16×10 -8 M, 8.70×10 -9 M or 2.15×10-7 M's K D To bind to mouse IL2Rγ; or to not bind in a detectable manner; approximately 3.32 × 10 at 25°C. -9 M ~ approx. 1.97×10 -7 M's K D To bind to human IL2Rγ domain 1; or to not bind in a detectable manner; approximately 4.13 × 10 at 37°C. -9 M ~ approx. 2.25×10 -7 M's K D To bind to human IL2Rγ domain 1; or to not bind in a detectable manner; approximately 2.91 × 10⁻¹⁴ at 25°C. -7 M ~ approx. 5.35×10 -10 K D To bind to human IL2Rγ domain 2; or to not bind in a detectable manner; approximately 1.14 × 10 at 37°C. -8 Alternatively, approximately 1.27 × 10 -8 K DTo bind to human IL2Rγ domain 2; or to fail to bind in a detectable manner; to block STAT phosphorylation in T cells induced by IL-2, IL-4, IL-7, IL-15 and / or IL-21; to block STAT phosphorylation in mast cells induced by IL-9; to reduce the number of human immune cells injected into mice; to reduce the levels of serum human cytokines and / or mouse serum cytokines in mice with human immune cells; to fail to bind to either mouse IL2Rγ or rat IL2Rγ in a detectable manner; to reduce body weight attributable to GvHD in a GvHD mouse model. The present invention provides isolated antigen-binding proteins (e.g., monospecific or multispecific antibodies or antigen-binding fragments thereof) characterized by one or more of the following: protecting mice from loss and / or death; blocking the binding of a hybrid receptor containing IL2Rγ complexed with a cytokine-specific receptor subunit to IL-2, IL-4, IL-7, IL-9, IL-15 and / or IL-21; and / or reducing the number of CD45+ cells, B cells, T cells and / or NK cells in the blood or serum of the subject (except in some cases, e.g., neutrophils). Antibodies and antigen-binding fragments whose sequence is a variant of any antibody or fragment particularly described herein and which specifically bind to IL2Rγ and are characterized by one or more of the traits described above form a part of the present invention.

[0011] The present invention also relates to (i) an isolated antigen-binding protein, such as an antibody, that specifically binds to the same epitope on IL2Rγ as a reference antibody or its antigen-binding fragment; or (ii) competes with the reference antibody or its antigen-binding fragment for binding to the IL2Rγ polypeptide. or an antigen-binding fragment thereof, wherein the reference antibody or its antigen-binding fragment contains (a) the amino acid sequence described in SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376; or a variant thereof, a heavy chain immunoglobulin or its variable region containing CDR-H1, CDR-H2 and CDR-H3. The present invention provides isolated antigen-binding proteins, such as antibodies or antigen-binding fragments thereof, comprising (b) the amino acid sequence described in SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378, or a variant thereof, comprising CDR-L1, CDR-L2 and CDR-L3 of the light chain immunoglobulin or its variable region. In embodiments of the present invention, the reference antibody or fragment is pre-bound to the IL2Rg antigen before the antigen-binding protein is added and evaluated for binding. In embodiments of the present invention, the antigen-binding protein is pre-bound to the antigen before a reference antibody or fragment is added and its binding is evaluated.

[0012] The present invention also relates to (a) an amino acid sequence described in SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376; or a heavy chain immunoglobulin or its variable region containing CDR-H1, CDR-H2 and CDR-H3. The present invention provides isolated antigen-binding proteins (e.g., antibodies or antigen-binding fragments thereof) comprising the amino acid sequence described in (b) SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378; or a light chain immunoglobulin

[0013] In embodiments of the present invention, the antigen-binding protein has at least 90% amino acid sequence identity with respect to the amino acid sequences described in (a) SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376. A heavy chain immunoglobulin or its variable region containing an amino acid sequence; and / or (b) a light chain immunoglobulin or its variable region containing an amino acid sequence having at least 90% amino acid sequence identity with respect to the amino acid sequences described in SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378. For example, in embodiments of the present invention, the antigen-binding proteins are (a) SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 2 The amino acid sequences described in 00, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376 or SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, Heavy chain immunoglobulin or its variable region containing CDR-H1, CDR-H2, and CDR-H3 of the variable region thereof, which contains an amino acid sequence having at least 90% amino acid sequence identity with the amino acid sequences described in 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376; (b) The amino acid sequences described in (b) SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378 or SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 12 A light chain immunoglobulin or a light chain immunoglobulin or its variable region containing CDR-L1, CDR-L2, and CDR-L3 of the variable region thereof, comprising an amino acid sequence having at least 90% amino acid sequence identity with respect to the amino acid sequences described in 7, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378.

[0014] In embodiments of the present invention, the antigen-binding protein is (i) Heavy chain CDR set: CDR-H1 containing the amino acid sequence described in SEQ ID NO: 4; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 6; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 8; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 24; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 26; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 28; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 44; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 46; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 48; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 64; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 66; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 68; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 83; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 85; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 87; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 103; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 105; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 107; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 121; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 123; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 125; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 140; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 142; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 144; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 158; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 160; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 162; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 176; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 178; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 180; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 192; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 194; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 196; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 202; and containing the amino acid sequence described in SEQ ID NO: 204 CDR-H2; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 206; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 176; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 212; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 214; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 220; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 222; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 224; and / or containing the amino acid sequence described in SEQ ID NO: 240 CDR-H1; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 242; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 244; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 260; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 262; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 264; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 278; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 280; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 282 ; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 288; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 290; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 292; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 298; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 300; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 302; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 317; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 319; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 321; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 337; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 339; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 341; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 347; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 349; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 351; and / or CDR-H1 containing the amino acid sequence described in SEQ ID NO: 363; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 66; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 366; and / or (ii) light chain CDR set: CDR-L1 containing the amino acid sequence described in SEQ ID NO: 12; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 14; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 16; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 32; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 34; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 36; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 52; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; CDR-L3 containing the amino acid sequence described in SEQ ID NO: 56; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; CDR-L3 containing the amino acid sequence described in SEQ ID NO: 75; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 91; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 93; CDR-L3 containing the amino acid sequence described in SEQ ID NO: 95; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 111; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; CDR-L3 containing the amino acid sequence described in SEQ ID NO: 113; and / or SEQ ID NO: 129 CDR-L1 containing the amino acid sequence described; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 132; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 148; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 150; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 166; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 14; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 168; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and containing the amino acid sequence described in SEQ ID NO: 184 CDR-L3; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 228; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 230; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 232; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 248; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 250; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 252; and / or containing the amino acid sequence described in SEQ ID NO: 268 CDR-L1; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 270; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 306; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 230; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 309; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 325; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 327; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 329; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; and CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54;and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 355; and / or CDR-L1 containing the amino acid sequence described in SEQ ID NO: 370; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 372; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 374; Includes.

[0015] In embodiments of the present invention, the antigen-binding protein of the present invention comprises the following heavy chain CDR set and light chain CDR set: (i) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 4; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 6; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 8; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 12; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 14; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 16; (ii) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 24; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 26; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 28; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 32; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 34; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 36; (iii) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 44; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 46; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 48; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 52; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 56; (iv) CD containing the amino acid sequence described in SEQ ID NO: 64; and CDR-H1 containing the amino acid sequence described in SEQ ID NO: 66 (v) Heavy chain variable region including R-H2; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 68; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 75; (v) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 83; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 85; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 87; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 91; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 93; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 95; (vi) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 103; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 105; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 107; and CDR-L1 containing the amino acid sequence described in SEQ ID NO: 111; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and (vi) Light chain variable region containing CDR-L3 containing the amino acid sequence described in sequence number 113; (vi) Heavy chain variable region containing CDR-H1 containing the amino acid sequence described in SEQ ID NO: 121; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 123; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 125; and Light chain variable region containing CDR-L1 containing the amino acid sequence described in SEQ ID NO: 129; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 132; (vii) SEQ ID NO: 140 (viii) CDR-H1 containing the amino acid sequence described; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 142; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 144; and the heavy chain variable region containing CDR-L1 containing the amino acid sequence described in SEQ ID NO: 148; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 150; (viii) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 158; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 160;(ix) Heavy chain variable region including CDR-H3 containing the amino acid sequence described in SEQ ID NO: 162; and CDR-L1 containing the amino acid sequence described in SEQ ID NO: 166; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 14; and light chain variable region including CDR-L3 containing the amino acid sequence described in SEQ ID NO: 168; (ix) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 176; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 178; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 180; and SEQ ID NO: Light chain variable region including CDR-L1 containing the amino acid sequence described in 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (x) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 192; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 194; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 196; and CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and (xi) Light chain variable region including CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (xi) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 202; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 204; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 206; and Light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (xii) SEQ ID NO: 176 (xiii) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 212; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 212; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 214; as well as a heavy chain variable region containing CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (xiii) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 220; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 222;and a heavy chain variable region including CDR-H3 containing the amino acid sequence described in SEQ ID NO: 224; and CDR-L1 containing the amino acid sequence described in SEQ ID NO: 228; sequence number; Light chain variable region including CDR-L2 containing the amino acid sequence described in SEQ ID NO. 230; and CDR-L3 containing the amino acid sequence described in SEQ ID NO. 232; (xiv) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO. 240; CDR-H2 containing the amino acid sequence described in SEQ ID NO. 242; and CDR-H3 containing the amino acid sequence described in SEQ ID NO. 244; and CDR-L1 containing the amino acid sequence described in SEQ ID NO. 248; CDR-L2 containing the amino acid sequence described in SEQ ID NO. 250; and CDR-L3 containing the amino acid sequence described in SEQ ID NO. 252. Light chain variable region including; (xv) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 260; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 262; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 264; and Light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 268; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 270; (xvi) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 278; Amino acid sequence described in SEQ ID NO: 280 Heavy chain variable region including CDR-H2 containing the sequence; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 282; and light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (xvii) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 288; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 290; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 292; and Light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 184; (xviii) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 298; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 300; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 302; as well as CDR-L1 containing the amino acid sequence described in SEQ ID NO: 306; and CDR-L2 containing the amino acid sequence described in SEQ ID NO: 230;and the light chain variable region including CDR-L3 containing the amino acid sequence described in SEQ ID NO: 309; (xix) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 317; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 319; and the heavy chain variable region including CDR-H3 containing the amino acid sequence described in SEQ ID NO: 321; as well as CDR-L1 containing the amino acid sequence described in SEQ ID NO: 325; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 327; and the light chain variable region including CDR-L3 containing the amino acid sequence described in SEQ ID NO: 329; (xx) CDR-H1 containing the amino acid sequence described in SEQ ID NO: 337; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 339; and the heavy chain variable region including CDR-H3 containing the amino acid sequence described in SEQ ID NO: 341; as well as CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and the amino acid sequence described in SEQ ID NO: 184 Light chain variable region including CDR-L3 containing the sequence; (xxi) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 347; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 349; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 351; and Light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 72; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 54; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 355; (xxii) Heavy chain variable region including CDR-H1 containing the amino acid sequence described in SEQ ID NO: 363; CDR-H2 containing the amino acid sequence described in SEQ ID NO: 66; and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 366; and Light chain variable region including CDR-L1 containing the amino acid sequence described in SEQ ID NO: 370; CDR-L2 containing the amino acid sequence described in SEQ ID NO: 372; and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 374.

[0016] A complex comprising the antigen-binding protein of the present invention bound to the IL2Rγ polypeptide or its antigenic fragment is also part of the present invention.

[0017] The present invention also relates to antigen-binding proteins (e.g., antibodies or their antigen-binding fragments) or their immunoglobulin chains (e.g., V H , V L The present invention provides a method for producing an antigen-binding protein (HC or LC), comprising: (a) introducing one or more polynucleotides (or a vector containing such polynucleotides) encoding one or more immunoglobulin chains of the antigen-binding protein into a host cell (e.g., a CHO cell); (b) culturing the host cell under conditions favorable for the expression of the polynucleotide; and (c) optionally isolating the antigen-binding protein or immunoglobulin chain from the host cell and / or the culture medium in which the host cell was grown. The antigen-binding protein or immunoglobulin chain that is a product of such a method also forms part of the present invention.

[0018] The present invention also relates to (a) heavy chain immunoglobulins or their variable regions CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences described in SEQ ID NOs: 2, 18, 22, 38, 42, 58, 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 and / or 376, or variants thereof; and / or (b) The present invention provides a polypeptide comprising the amino acid sequence described in SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 and / or 378, or a variant thereof, in a light chain immunoglobulin or its variable region CDR-L1, CDR-L2, and CDR-L3; or (c) a member selected from the group consisting of SEQ ID NOs: 1 to 378, or a variant thereof. The present invention also provides a polynucleotide encoding one or more such polypeptides or a vector (e.g., plasmid) comprising such polynucleotides.

[0019] The present invention also relates to antigen-binding proteins (e.g., antibodies or antigen-binding fragments thereof) or immunoglobulin chains (e.g., V) as described herein. H , V L The present invention provides a host cell (e.g., a CHO cell) comprising a polypeptide (HC or LC) or polynucleotide or vector.

[0020] The present invention also provides compositions or kits comprising one or more of the antigen-binding proteins described herein (e.g., antibodies or antigen-binding fragments thereof), optionally combined with further therapeutic agents (e.g., anti-inflammatory agents, anti-TNFα antibodies or binding proteins, infliximab, adalimumab, etanercept, golimumab, corticoids, prednisolone, methylprednisolone, anti-thymocyte globulin, alemtuzumab, daclizumab, in vitro photopheresis, mycophenolate mofetil, sirolimus, pentostatin, mesenchymal stem cells, inorimomab, denileukin, or basiliximab).

[0021] The present invention further provides pharmaceutical formulations comprising an antigen-binding protein described herein (e.g., an antibody or its antigen-binding fragment) and a pharmaceutically acceptable carrier, and optionally further therapeutic agents (e.g., an anti-inflammatory agent, an anti-TNFα antibody or binding protein, infliximab, adalimumab, etanercept, golimumab, a corticoid, prednisolone, methylprednisolone, anti-thymocyte globulin, alemtuzumab, daclizumab, in vitro photopheresis, mycophenolate mofetil, tacrolimus, cyclosporine, sirolimus, pentostatin, mesenchymal stem cells, inorimomab, denileukin, or basiliximab).

[0022] The present invention also relates to antigen-binding proteins or compositions described herein (e.g., pharmaceutical formulations). The present invention provides containers or injection devices (e.g., vials, syringes, pre-filled syringes, or auto-injectors) that include ).

[0023] The present invention also provides a method for administering an antigen-binding protein or composition described herein to a subject (e.g., a human), comprising introducing the antigen-binding protein or composition into the subject's body, for example, by injection (e.g., subcutaneously, intravenously, or intramuscularly). The present invention also provides a method for treating or preventing an IL2Rγ-mediated disease or condition (e.g., graft-versus-host disease, organ graft rejection, skin graft rejection, heart graft rejection, lung graft rejection, kidney graft rejection, liver graft rejection, scatter chorioretinopathy, multiple sclerosis, uveitis, autoimmune diseases, type 1 diabetes, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, and / or myasthenia gravis) in a subject requiring such treatment, comprising administering, for example, by injection, an effective amount of an antigen-binding protein or composition described herein.

[0024] The present invention also aims to block STAT phosphorylation in peripheral blood mononuclear cells (e.g., T cells) induced by cytokines (e.g., IL-2, IL-4, IL-7, IL-15, and / or IL-21); block STAT (e.g., STAT3) phosphorylation in mast cells induced by cytokines (e.g., IL-9); reduce serum levels of interferon-gamma, tumor necrosis factor-alpha, IL-6, IL-8, IL-10, and / or mKC / GRO (e.g., in transplanted subjects); and the IL2Rγ family. The present invention provides a method for blocking JAK-STAT-mediated (e.g., STAT3) intracellular signaling (e.g., in NK cells) induced by cytokines (e.g., IL-2, IL-4, IL-7, IL-9, IL-15, and / or IL-21); and / or for reducing serum levels of CD45+ immune cells, NK cells, T cells, and / or B cells (e.g., excluding neutrophils) in a subject, comprising administering to the subject an effective amount of the anti-IL2Rγ antigen-binding protein or a composition or preparation thereof described herein. In embodiments of the present invention, the subjects are those suffering from IL2Rγ-mediated diseases or conditions, such as graft-versus-host disease, organ graft rejection, β-islet cell graft rejection, skin graft rejection, heart graft rejection, lung graft rejection, kidney graft rejection, liver graft rejection, shotgun chorioretinopathy, multiple sclerosis, uveitis, autoimmune diseases, type 1 diabetes, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, myasthenia gravis, aplastic anemia, atopic dermatitis, asthma, mast cell activation disorder, mast cell activation syndrome (MCAS), systemic hypertrophic cell disease (SM), and / or mast cell leukemia (MCL). [Brief explanation of the drawing]

[0025] [Figure 1A]Figure 1(A) shows the blockade of human (A)IL-2, (B)IL-4, (C)IL-7, (D)IL-15, and (E)IL-21-induced STAT phosphorylation in human CD4+ T cells by various concentrations of anti-IL-2R gamma antibodies H4H12857P, H4H12874P, H4H12886P, H4H12889P, and H4H12922P2; as well as antibodies REGN1945 and COMP1499. [Figure 1B] Figure 1(B) shows the blockade of human (A)IL-2, (B)IL-4, (C)IL-7, (D)IL-15, and (E)IL-21-inducible STAT phosphorylation in human CD4+ T cells by various concentrations of anti-IL-2R gamma antibodies H4H12857P, H4H12874P, H4H12886P, H4H12889P, and H4H12922P2; as well as antibodies REGN1945 and COMP1499. [Figure 1C] Figure 1(C) shows the blockade of human (A)IL-2, (B)IL-4, (C)IL-7, (D)IL-15, and (E)IL-21-inducible STAT phosphorylation in human CD4+ T cells by various concentrations of anti-IL-2R gamma antibodies H4H12857P, H4H12874P, H4H12886P, H4H12889P, and H4H12922P2; as well as antibodies REGN1945 and COMP1499. [Figure 1D] Figure 1(D) shows the blockade of human (A)IL-2, (B)IL-4, (C)IL-7, (D)IL-15, and (E)IL-21-inducible STAT phosphorylation in human CD4+ T cells by various concentrations of anti-IL-2R gamma antibodies H4H12857P, H4H12874P, H4H12886P, H4H12889P, and H4H12922P2; as well as antibodies REGN1945 and COMP1499. [Figure 1E] Figure 1(E) shows the blockade of human (A)IL-2, (B)IL-4, (C)IL7, (D)IL-15, and (E)IL-21-inducible STAT phosphorylation in human CD4+ T cells by various concentrations of anti-IL-2R gamma antibodies H4H12857P, H4H12874P, H4H12886P, H4H12889P, and H4H12922P2; as well as antibodies REGN1945 and COMP1499. [Figure 2] Blocking of human IL-9-induced STAT3 phosphorylation in vitro differentiated human mast cells by anti-IL-2R gamma antibodies H4H12874P, H4H12886P, H4H12889P, H4H12922P2; and antibodies COMP1499 and REGN1945. [Figure 3] Figure 3(A-F) shows the percentage of initial body weight over time in mice with human PBMCs that were administered the anti-IL2R gamma antibodies (E)H4H12889P and (F)H4H12922P2, as well as antibody (D)COMP1499. Control experiments in mice that were not administered antibodies (B), (C) administered isotype control antibodies, or (A) lack human PBMCs are also shown. The start of antibody injection at day 21 and the end of antibody injection at day 59 are indicated by dashed lines. [Figure 4] This shows the time course of survival in mice injected with anti-IL2R gamma antibodies H4H12889P and H4H12922P2, antibody COMP1499, antibody REGN1945, and mice that were not injected with antibodies. The uninjected huPBMC group is not depicted. Differences in animal survival compared to the isotype control antibody group were analyzed by the Mantell-Cox log-rank test. A p-value < 0.05 was considered statistically significant. **, p-value < 0.0021; ****, p-value < 0.0001. The start of antibody injection at day 21 and the end of antibody injection at day 59 are indicated by dashed lines. [Figure 5]Figure 5(A-D) shows the absolute human cell count in the blood of mice 35 days after huPBMC injection in mice that were not administered an antibody (IgG-free) or were administered REGN1945, COMP1499, or the anti-IL2R gamma antibody H4H12889P or H4H12922P2 ((A) human CD45 cells; (B) human T cells; (C) human CD4 T cells; and (D) human CD8 T cells). The group "huPBMC-free" is not shown; #, significantly different from the group "huPBMC-free"; †, significantly different from the group "huPBMC-IgG-free"; *, significantly different from the group "huPBMC-REGN1945". Each symbol represents a mouse. A value of 0 was arbitrarily changed by 0.01 for the purpose of graph creation (logarithmic scale). [Figure 6] Figure 6(A-D) shows the blood cell counts of human (A) CD45+ cells, (B) T cells, (C) CD4+ T cells, and (D) CD8+ T cells over time in mice administered with anti-IL2R gamma antibodies H4H12889P or H4H12922P2; or COMP1499 or isotype control antibodies. The start of antibody injection on day 21 and the end of antibody injection on day 59 are indicated by dashed lines. [Figure 7-1]Figure 7(A) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "anti-huPBMC" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. Figure 7(B) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "huPBMC-free" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. [Figure 7-2]Figure 7(C) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "anti-huPBMC" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. Figure 7(D) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "huPBMC-free" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. Figure 7(E) shows the serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs.# indicates a significant difference from the "huPBMC-free" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. [Figure 7-3] Figure 7(F) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "huPBMC-free" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. Figure 7(G) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "huPBMC-free" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. [Figure 7-4]Figure 7(H) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "anti-huPBMC" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. Figure 7(I) shows serum levels of human and mouse cytokines ((A) human interferon-gamma; (B) human TNFα; (C) human IL-6; (D) human IL-8; (E) human IL-10; (F) mouse TNFα; (G) mouse IL-6; (H) mouse KC / GRO; and (I) mouse IL-10) 42 days after huPBMC injection in mice that were not administered antibodies (IgG-free), or that were administered REGN1945, COMP1499, or anti-IL2R gamma antibodies H4H12889P or H4H12922P2, or mice without human PBMCs. # indicates a significant difference from the "anti-huPBMC" group; † indicates a significant difference from the "huPBMC-IgG-free" group; * indicates a significant difference from the "huPBMC-REGN1945" group. Each symbol represents a mouse. [Figure 8] Figure 8(A-D) shows the serum levels of (A) human IFN-γ, (B) human TNFα, (C) mouse TNFα, and (D) mouse IL-6 over time in mice administered with anti-IL2R gamma antibodies H4H12889P or H4H12922P2; or COMP1499 or isotype control antibodies. [Figure 9A] Figure 9(A) shows the levels of total human antibody or CD45+ immune cells (A), NK cells (B), T cells (C), B cells (D), or neutrophils (E) in the blood of mice treated with various doses of the antibody REGN1945 or H4H12889P. [Figure 9B] Figure 9(B) shows the levels of total human antibody or CD45+ immune cells (A), NK cells (B), T cells (C), B cells (D), or neutrophils (E) in the blood of mice treated with various doses of the antibody REGN1945 or H4H12889P. [Figure 9C] Figure 9(C) shows the levels of total human antibody or CD45+ immune cells (A), NK cells (B), T cells (C), B cells (D), or neutrophils (E) in the blood of mice treated with various doses of the antibody REGN1945 or H4H12889P. [Figure 9D] Figure 9(D) shows the levels of total human antibody or CD45+ immune cells (A), NK cells (B), T cells (C), B cells (D), or neutrophils (E) in the blood of mice treated with various doses of the antibody REGN1945 or H4H12889P. [Figure 9E] Figure 9(E) shows the levels of total human antibody or CD45+ immune cells (A), NK cells (B), T cells (C), B cells (D), or neutrophils (E) in the blood of mice treated with various doses of the antibody REGN1945 or H4H12889P. [Figure 10] Experimental design for in vivo skin graft rejection experiments. [Figure 11] Time to onset of skin graft rejection in mice that were not administered antibodies or were administered REGN1945 or H4H12889P. [Figure 12] Time to complete rejection of skin grafts in mice that were not administered antibodies or were administered REGN1945 or H4H12889P. [Figure 13] Total donor-specific IgG antibodies in non-transplanted or transplanted mice that were not administered antibodies, or were administered REGN1945 or H4H12889P. [Modes for carrying out the invention]

[0026] The present invention provides an antibody and its antigen-binding fragment that specifically binds to human and cynomolgus monkey IL2Rγ and exhibits exceptional biological activity, particularly in blocking cytokine-induced STAT phosphorylation in T cells and blocking graft-versus-host disease in applicable mouse models.

[0027] According to the present invention, conventional molecular biology, microbiology, and recombinant DNA techniques within the technical scope of the art can be used. Such techniques are well described in the literature. For example, Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual, Second Edition(1989)Cold Spring Harbor Laborat IRL Press, Cold Spring Harbor, NY (referred to herein as "Sambrook et al., 1989"); DNA Cloning: A Practical Approach, Volume I and II (edited by DNGlover, 1985); Oligonucleotide Synthesis (edited by MJ Gait, 1984); Nucleic Acid Hybridization (edited by B.D. Hames & S.J. Higgins, (1985)); Transcription and Translation (edited by B.D. Hames & S.J. Higgins, (1984)); Animal Cell Culture (edited by R. Freshney, (1986)); Immobilized Cells and Enzymes (IRL Press, (1986)); B. Perbal, A Practical Guide See To Molecular Cloning (1984); FMAusubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, Inc. (1994).

[0028] IL-2Rγ The interleukin-2 receptor subunit gamma is also known as CD132; common cytokine receptor γc chain; IL-2RG; IL-2Rg; IL2Rgamma; IL-2Rγ, IMD4; P64:SCIDX; or SCIDX1. IL2Rγ is a common subunit for several interleukin receptors, including IL-2R, IL-4R, IL-7R, IL-9R, IL-15R, and IL21R.

[0029] In embodiments of the present invention, human IL2Rγ is encoded by the nucleotide sequence described under Genbank accession number NM_000206. In embodiments of the present invention, human IL2Rγ comprises the amino acid sequence described under Genbank accession number NP_000197.

[0030] Antigen-binding protein The present invention provides antigen-binding proteins, such as antibodies (e.g., human antibodies, monoclonal antibodies, and recombinant antibodies) and their antigen-binding fragments, that specifically bind to the IL2Rγ protein or its antigenic fragment (e.g., the extracellular domain of IL2Rγ). Antigen-binding proteins that bind to the same epitope on IL2Rγ as any of the antigen-binding proteins described herein, or that compete with any of the antigen-binding proteins described herein for binding to IL2Rγ, are also part of the present invention.

[0031] The present invention also relates to Sequence IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 1 23, 125, 127, 129, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 19 2, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, The following provides any polypeptide containing the amino acid sequences or variants thereof described in 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 366, 368, 370, 372, 374, 376 and / or 378. In some cases, the polypeptide is fused with one or more other polypeptides, such as human Fc (e.g., human IgG, e.g., IgG1 or IgG4 (e.g., including the S108P mutation)).

[0032] When used herein, the term "antibody" refers to an immunoglobulin molecule (i.e., a "complete antibody molecule") (e.g., IgG) comprising four polypeptide chains, two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds, such as H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P; H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2. In embodiments of the present invention, each antibody heavy chain (HC) has a heavy chain variable region ("HCVR" or "V H )(for example, SEQ ID NOs. 2, 22, 42, 62, 81, 101, 119, 138, 156, 174, 190, 200, 210, 218, 238, 258, 276, 286, 296, 315, 335, 345 or 361 or their variants) and heavy chain constant region (domain C H 1, C H 2 and C H Includes 3); each antibody light chain (LC) has a light chain variable region ("LCVR" or "V"). L )(for example, SEQ ID NOs. 10, 30, 50, 70, 89, 109, 127, 146, 164, 182, 226, 246, 266, 304, 323, 353 or 368 or their variants) and light chain constant region (C L ) includes. V H and V L The domain can be further divided into a more conserved domain called the framework domain (FR) and a highly variable domain called the complementarity determination domain (CDR). H and V LThis comprises three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments of the present invention, the FRs of the antibody (or its antigen-binding fragment) are identical to those of the human germline sequence, or are naturally or artificially modified.

[0033] Typically, the variable domains of both heavy and light immunoglobulin chains contain three hypervariable regions, also called complementarity-determining regions (CDRs), located within a relatively conserved framework region (FR). Generally, from the N-terminus to the C-terminus, the variable domains of both the light and heavy chains include FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. In embodiments of the present invention, the assignment of amino acids to each domain is given by Sequences of Proteins of Immunological Interest, Kabat et al; National Institutes of Health, Beth esda, Md.;5th ed.;NIH Publ.No.91~3242(1991);Kabat(1978)Adv.Prot.Chem.32:1~75;Kab at et al., (1977) J. Biol. Chem. 252: pp. 6609-6616; Cho thia et al., (1987) J Mol. Biol. 196: pp. 901-917 or Ch According to the definition by Othia et al. (1989) Nature 342:878-883. Therefore, the present invention is V H CDR and V L An antibody and antigen-binding fragment containing a CDR of V H and V L The antibody and antigen-binding fragments include an amino acid sequence (or a variant thereof) as described herein, and the CDR is defined according to Kabat and / or Chothia.

[0034] When used herein, terms such as “antigen-binding portion” or “antigen-binding fragment” of an antibody or antigen-binding protein include any naturally occurring, enzymatically obtainable, synthetic, or genetically modified polypeptide or glycoprotein that specifically binds to an antigen and forms a complex. Non-limiting examples of antigen-binding fragments include (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments (heavy chain portion of a Fab fragment cleaved with papain); (iv) Fv fragments (V H or V L (v) single-chain Fv(scFv) molecules; these include those consisting of amino acid residues that mimic the hypervariable region of an antibody (e.g., isolated complementarity-determining regions (CDRs) such as the CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other manipulated molecules, such as domain-specific antibodies, single-domain antibodies, domain deletion antibodies, chimeric antibodies, CDR graft antibodies, diabodies, triabodies, tetrabodies, minibodies, and small modular immunotherapy drugs (SMIPs), are also included within the expression “antigen-binding fragment” as used herein. In the embodiments of the present invention, the antigen-binding fragments are H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H129 Includes three or more CDRs of the following types: 22P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2; or H4H13545P2 (e.g., CDR-H1, CDR-H2 and CDR-H3; or CDR-L1, CDR-L2 and CDR-L3).

[0035] In embodiments of the present invention, the antigen-binding protein of the present invention (e.g., an antibody or its antigen-binding fragment) is as shown in Table A below: [Table 1] V containing the heavy chain CDR combinations described above (CDR-H1, CDR-H2, and CDR-H3) H Heavy chain immunoglobulins (e.g., HC) containing, and / or Table B below: [Table 2] V containing the combination of light chain CDRs described above (CDR-L1, CDR-L2, and CDR-L3) L It contains light chain immunoglobulins (e.g., LC).

[0036] In embodiments of the present invention, the antigen-binding protein of the present invention (e.g., an antibody or its antigen-binding fragment) is as follows: Table C: [Table 3] V includes combinations of heavy and light chain CDRs as described above (CDR-H1, CDR-H2 and CDR-H3; as well as CDR-L1, CDR-L2 and CDR-L3). H and V L It includes heavy chain (e.g., HC) and light chain (e.g., LC) immunoglobulins, each containing the respective components.

[0037] The present invention relates to the following V H and V L Includes antigen-binding proteins (e.g., antibodies or their antigen-binding fragments) containing polypeptide pairs with amino acid sequences: Sequence IDs 2 and 10; Sequence IDs 22 and 30; Sequence IDs 42 and 50; Sequence IDs 62 and 70; Sequence IDs 81 and 89; Sequence IDs 101 and 109; Sequence IDs 119 and 127; Sequence IDs 138 and 146; Sequence IDs 156 and 164; Sequence IDs 174 and 182; Sequence IDs 190 and 182; Sequence IDs 200 and 182; Sequence IDs 210 and 182; Sequence IDs 218 and 226; Sequence IDs 238 and 246; Sequence IDs 258 and 266; Sequence IDs 276 and 182; Sequence IDs 286 and 182; Sequence IDs 296 and 304; Sequence IDs 315 and 323; Sequence IDs 335 and 182; Sequence IDs 345 and 353; or Sequence IDs 361 and 368.

[0038] The present invention includes an antigen-binding protein (e.g., an antibody or its antigen-binding fragment) comprising the following amino acid sequence pairs encoding HC and LC: Sequence IDs 18 and 20; Sequence IDs 38 and 40; Sequence IDs 58 and 60; Sequence IDs 77 and 79; Sequence IDs 97 and 99; Sequence IDs 115 and 117; Sequence IDs 134 and 136; Sequence IDs 152 and 154; Sequence IDs 170 and 172; Sequence IDs 186 and 188; Sequence IDs 198 and 188; Sequence IDs 208 and 188; Sequence IDs 216 and 188; Sequence IDs 234 and 236; Sequence IDs 254 and 256; Sequence IDs 272 and 274; Sequence IDs 284 and 188; Sequence IDs 294 and 188; Sequence IDs 311 and 313; Sequence IDs 331 and 333; Sequence IDs 343 and 188; Sequence IDs 357 and 359; or Sequence IDs 376 and 378.

[0039] Embodiments of the present invention are also the corresponding V described in particular herein. H , V L Having 70% or higher (e.g., 80%, 85%, 90%, 95%, 97%, or 99%) overall amino acid sequence identity or similarity to the amino acid sequence of HC or LC, however such immunoglobulins CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 are not variants, and immunoglobulin V containing variant amino acid sequences including the amino acid sequences described herein. H and V L ; or an antigen-binding protein comprising HC and LC, for example, an anti-IL2Rγ antibody and its antigen-binding fragment. Thus, in such embodiments, the CDRs within the variant antigen-binding protein are themselves not variants.

[0040] The present invention includes a monoclonal composition comprising a plurality of isolated monoclonal antigen-binding proteins, in addition to a monoclonal anti-IL2Rγ antigen-binding protein, such as an antibody and its antigen-binding fragment. The terms "monoclonal antibody" or "mAb," as used herein, refer to a member of a substantially homogeneous population of antibodies, i.e., a cluster. The antibody molecules constituting the group are identical in amino acid sequence, except for possible naturally occurring mutations that may be present in trace amounts. “Multiple” such monoclonal antibodies and fragments in a composition refer to concentrations of identical antibodies and fragments (i.e., identical in amino acid sequence, except for possible naturally occurring mutations that may be present in trace amounts, as discussed above) that are higher than what would normally occur in nature, for example, in the blood of a host organism such as a mouse or a human.

[0041] In embodiments of the present invention, the anti-IL2Rγ antigen-binding protein, for example, an antibody or antigen-binding fragment, comprises a heavy chain constant domain, for example, an IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3, and IgG4 (e.g., including S228P and / or S108P mutations)) or IgM type heavy chain constant domain. In embodiments of the present invention, the antigen-binding protein, for example, an antibody or antigen-binding fragment, comprises a light chain constant domain, for example, a kappa or lambda type light chain constant domain. The present invention relates to antigen-binding proteins comprising variable domains described herein, such as those described above, which are linked to heavy chain and / or light chain constant domains (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12 Includes 889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2).

[0042] The term “human” antigen-binding protein, e.g., antibody or antigen-binding fragment, as used herein, includes antibodies and fragments having variable and constant regions derived from human germline immunoglobulin sequences, whether in human cells or transplanted into non-human cells, e.g., mouse cells. See, for example, US8502018, US6596541, or US5789215. In embodiments of the present invention, the human antibodies and antigen-binding fragments may include, for example, CDRs, particularly CDR3, amino acid residues not encoded by human germline immunoglobulin sequences (e.g., having mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term “human antibody” is not intended to include mAbs in which a CDR sequence derived from the germline of another mammalian species (e.g., mouse) is grafted onto a human FR sequence. The term includes antibodies produced by recombination in or within non-human mammals. The term is not intended to include antibodies isolated from or generated in human subjects. This invention relates to human antigen-binding proteins (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; This includes antibodies or antigen-binding fragments such as H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2.

[0043] The present invention comprises anti-IL2Rγ chimeric antigen-binding proteins, such as antibodies and their antigen-binding fragments, and methods of using them. As used herein, "chimeric antibody" is an antibody having a variable domain from a first antibody and a constant domain from a second antibody, wherein the first and second antibodies are from different species. (e.g., US4816567; and Morrison et al., (1984) Proc. Natl. Aca See d.Sci.USA 81:6851-6855). The present invention is (for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P; A chimeric antibody comprising the variable domains described herein (from H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2).

[0044] The term "recombinant" antigen-binding protein, e.g., an antibody or an antigen-binding fragment thereof, refers to a molecule made, expressed, isolated or obtained by a technique or method known in the art such as recombinant DNA technology, including, e.g., DNA splicing and transgenic expression. The term includes antibodies expressed in a non-human mammal (including transgenic non-human mammals, e.g., transgenic mice), or a host cell (e.g., Chinese hamster ovary (CHO) cells) or a cell expression system, or isolated from a recombinant combinatorial human antibody library. The present invention includes recombinant antigen-binding proteins as described herein (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P; H4H12913P2; H4H12922P2; H4H12924P2; H4H12926P2; H4H12927P2; H4H12934P2; H4H13538P; H4H13541P; H4H13544P2; or H4H13545P2).

[0045] An antigen-binding fragment of an antibody, in an embodiment of the present invention, includes at least one variable domain. The variable domain can be of any size or amino acid composition and generally includes at least one (e.g., three) CDRs adjacent to or in-frame with one or more framework sequences. The V L domain associated with the V H domain in an antigen-binding fragment having a V H and V L domain can be positioned in any suitable arrangement relative to each other. For example, the variable region can be a dimer and can contain a V H -V H , V H -V L or V L -V L dimer. Alternatively, an antigen-binding fragment of an antibody can be a non-covalently bound monomeric VH and / or V L may contain a domain.

[0046] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found within the antigen-binding fragments of the antibodies of the present invention include: (i) V H -C H 1; (ii) V H -C H 2; (iii) V H -C H 3; (iv) V H -C H 1-C H 2; (v) V H -C H 1-C H 2-C H 3; (vi) V H -C H 2-C H 3; (vii) V H -C L ; (viii) V L -C H 1; (ix) V L -C H 2; (x) V L -CH3; (xi) V L -C H 1-C H 2; (xii) V L -C H 1-C H 2-C H 3; (xiii) V L -C H 2-C H 3; and (xiv) V L -C LExamples include: In any configuration of variable and constant domains, including any exemplary configurations listed above, the variable and constant domains may be directly linked to each other or linked by a whole or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids resulting in a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody of the present invention may be non-covalently associated with and / or (e.g., by disulfide bonds) one or more monomers V H Or V L Homodimers or heterodimers of any variable and constant domain configurations listed above with respect to the domain ( The present invention may also include other multimers. The present invention relates to antigen-binding proteins as described herein, for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4 Contains antigen-binding fragments of H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2; or H4H13545P2.

[0047] Antigen-binding proteins (e.g., antibodies and antigen-binding fragments) may be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding proteins are discussed further herein. The present invention relates to antigen-binding proteins particularly described herein (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P; H4H1 In addition to monospecific antigen-binding fragments containing one or more variable domains from 2913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2), they also contain multispecific (e.g., bispecific) antigen-binding fragments.

[0048] The terms "specifically binds" or "binds specifically" refer to real-time label-free biolayer interferometry assays at 25°C or 37°C, e.g., using the Octet® HTX biosensor, or surface plasmon resonance, e.g., BI. At least about 10 as measured by ACORE (trademark) or by solution affinity ELISA. -7 M (for example, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 M) of, K DThis refers to an antigen-binding protein (e.g., an antibody or its antigen-binding fragment) that has binding affinity to an antigen such as the IL2Rγ protein, as expressed as . The present invention includes an antigen-binding protein that specifically binds to the IL2Rγ protein. In embodiments of the present invention, the anti-IL2Rγ antigen-binding protein is K for binding to human and / or mouse and / or cynomolgus monkey and / or rat IL2Rγ or their domains, as described in any of Tables 3-1 to 3-12. D Includes values. "Anti-IL2R gamma" refers to an antigen-binding protein (or other molecule) that specifically binds to IL2R gamma, such as an antibody or its antigen-binding fragment.

[0049] "Isolated" antigen-binding proteins (e.g., antibodies or their antigen-binding fragments), polypeptides, polynucleotides, and vectors do not contain, at least partially, other biological molecules from the cells or cell cultures from which their production originates. Such biological molecules include nucleic acids, proteins, other antibodies or antigen-binding fragments, lipids, carbohydrates, or other materials, e.g., cell debris and growth media. Isolated antigen-binding proteins may also not contain, at least partially, expression system components, e.g., biological molecules from host cells or their growth media. In general, the term "isolated" is not intended to mean the complete absence of such biological molecules (e.g., trace or insignificant amounts of impurities may remain), or the absence of water, buffers, or salts, or components of a pharmaceutical formulation containing antigen-binding proteins (e.g., antibodies or antigen-binding fragments).

[0050] The present invention relates to antigen-binding proteins (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H128 This includes antigen-binding proteins, such as antibodies or antigen-binding fragments, that bind to the same epitopes as 71P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2).

[0051] An antigen is, for example, a molecule to which an antibody binds, such as a peptide (e.g., IL2R gamma or a fragment thereof (antigenic fragment)). The specific region on an antigen that an antibody recognizes and binds to is called an epitope. The antigen-binding protein of the present invention (e.g., an antibody) that specifically binds to such an antigen is part of the present invention.

[0052] The term “epitope” refers to an antigenic determinant (e.g., on IL2Rγ) that interacts with a specific antigen-binding site on an antigen-binding protein, known as a paratope, for example, a variable region of an antibody molecule. A single antigen may have more than one epitopes. Thus, different antibodies may bind to different compartments on an antigen and have different biological effects. The term “epitope” can also refer to a site on an antigen to which B and / or T cells respond, and / or a region of the antigen to which an antibody binds. Epitopes can be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and have residues that directly contribute to the affinity of the interaction. Epitopes may be linear or conformal, i.e., composed of non-linear amino acids. In certain embodiments, epitopes may include determinants, which are chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and in certain embodiments, they may have distinctive three-dimensional structural features and / or distinctive electrical features. The epitope to which the antigen-binding protein of the present invention binds can be contained in a fragment of IL2Rγ, for example, human IL2Rγ, for example, in its ectodomain, domain 1, or domain 2. The antigen-binding protein of the present invention (e.g., an antibody) that binds to such an epitope is part of the present invention.

[0053] Methods for determining the epitopes of antigen-binding proteins, such as antibodies, fragments, or polypeptides, include alanine scanning mutation analysis and peptide blot analysis (Reineke (2004) Methods Mol. Biol. 248: pp. 443-463). Examples include peptide cleavage analysis, crystallographic studies, and NMR analysis. Additionally, methods such as epitope excision, epitope extraction, and chemical modification of antigens can be used (Tomer (2000) Prot.Sci.9: pp. 487-496). Antigen-binding proteins (for example) Another method that can be used to identify amino acids within a polypeptide that interact with an antibody, fragment, or polypeptide is hydrogen / deuterium exchange detected by mass spectrometry. For example, Ehring (1999) Analytical Biochem istry 267: pp. 252-259; Engen and Smith (2001) An See al.Chem.73:256A~265A.

[0054] The present invention relates to antigen-binding proteins, for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H 4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 and antigen-binding proteins that compete for binding to IL2Rγ, for example, variant IL2Rγ epitopes as discussed herein. The term “competing” is used herein when An antigen-binding protein (e.g., an antibody or its antigen-binding fragment) binds to an antigen (e.g., IL2Rγ) and inhibits or blocks the binding of another antigen-binding protein (e.g., an antibody or its antigen-binding fragment) to that antigen. Unless otherwise stated, the term also includes competition between two antigen-binding proteins in both directions, e.g., antibodies, i.e., a first antibody binds to an antigen and blocks binding by a second antibody, and vice versa. Thus, in embodiments of the present invention, competition occurs in one such direction. In certain embodiments, the first antigen-binding protein (e.g., an antibody) and the second antigen-binding protein (e.g., an antibody) may bind to the same epitope. Alternatively, the first and second antigen-binding proteins (e.g., antibodies) may bind to different but overlapping or non-overlapping epitopes, where one binding inhibits or blocks the binding of the second antibody, e.g., via steric hindrance. Competition among antigen-binding proteins (e.g., antibodies) can be measured by methods known in the art, for example, by a real-time label-free biolayer interferometry assay. Furthermore, binding competition among anti-IL2Rγ antigen-binding proteins (e.g., monoclonal antibodies (mAbs)) can be determined using a real-time label-free biolayer interferometry assay on an Octet RED384 biosensor (Pall ForteBio Corp.).

[0055] Typically, an antibody or antigen-binding fragment of the present invention, modified in any way, retains the ability to specifically bind to IL2Rγ, for example, retaining at least 10% of its IL2Rγ-binding activity when its activity is expressed on a molar basis (compared to the parent antibody). Preferably, the antibody or antigen-binding fragment of the present invention retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the IL2Rγ-binding affinity of the parent antibody. The antibody or antigen-binding fragment of the present invention is also intended to contain conserved or non-conserved amino acid substitutions (referred to as “conserved variants” or “functionally conserved variants” of the antibody) that do not substantially alter its biological activity.

[0056] polypeptides such as immunoglobulin chains (for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P) containing amino acid sequences specifically described herein. ;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 V H , V LA "variant" of HC or LC or its CDR is defined as the reference amino acid sequence described herein (e.g., SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, This refers to polypeptides containing at least approximately 70–99.9% (for example, at least 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, or 99.9%) identical or similar amino acid sequences to any of 355, 357, 359, 361, 363, 366, 368, 370, 372, 374, 376, or 378.

[0057] Furthermore, a polypeptide variant may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) mutations, e.g., one or more missense mutations (e.g., conservative substitutions), nonsense mutations, deletions, or insertions, except for a polypeptide such as an immunoglobulin chain (e.g., H4H12857P; H4H12858P; H4H12859) whose amino acid sequence is that of a reference polypeptide specifically described herein. P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H1 2913P2; H4H12922P2; H4H12924P2; H4H12926P2; H4H12927P2; H4H12934P2; H4H13538P; H4H13541P; V H , V L The present invention may include an immunoglobulin light chain (or V) containing the amino acid sequence described in SEQ ID NO: 10 but having one or more such mutations.L ) Immunoglobulin heavy chains (or V) containing the amino acid sequence described in SEQ ID NO: 2 but having one or more such mutations H The anti-IL2Rγ antigen-binding protein comprises an immunoglobulin light chain variant containing such a CDR, in which one or more (e.g., one, two, or three) of CDR-L1, CDR-L2, and CDR-L3 have one or more such mutations (e.g., conservative substitutions), and / or an immunoglobulin heavy chain variant containing such a CDR, in which one or more (e.g., one, two, or three) of CDR-H1, CDR-H2, and CDR-H3 have one or more such mutations (e.g., conservative substitutions).

[0058] The following reference concerns the BLAST algorithm, which is often used for sequence analysis: BLAST algorithm: Altschul et al. (2005) FEBS J.2 72(20):pp. 5101-5109; Altschul, SF et al., (1990) J .Mol.Biol.215:pp. 403-410; Gish, W. et al., (1993)Na ture Genet.3:pp. 266-272; Madden, TL et al., (1996) Meth. Enzymol.266:pp. 131-141; Altschul, SF (1997) Nucleic Acids Res. 25: pp. 3389-3402; Zhang, J. et al., (1997) Genome Res. 7: pp. 649-656; Wo Otton, JC et al., (1993) Comput. Chem. 17:149~163 Page; Hancock, JM et al., (1994) Comput. Appl. Biosc. .10:67~70; Alignment scoring system: Dayhoff, MO et al., "A model of evolutionary change in proteins," Atlas of Protein Sequence and Structure, (1978) vol.5, suppl.3. MO Dayhoff (Eds.), pp. 345-352, Natl. Biomed. Res. Found., Washington, DC; Schwartz, RM et al., "Matrices for detecting distant relationships.", Atlas of Protein Sequence and Structure, (1978) vol.5, suppl.3.” MODayhoff (ed.), pp. 353-358, Natl. Biomed. Res. Found., Washington, DC; Altschul, SF, (1991) J. Mol. Biol. 219: 555-56 Page 5; States, DJ et al. (1991) Methods 3:66-70; H Enikoff, S. et al. (1992) Proc. Natl. Acad. Sci. USA 89: pp. 10915-10919; Altschul, SF et al., (1993) J. Mol.Evol.36:pp. 290-300; Alignment statistics: Karlin, S. et al., (1990) Proc.Natl.Acad.Sci.USA 87:2264-22 Page 68; Karlin, S. et al., (1993) Proc. Natl. Acad. Sci. USA 90: pp. 5873-5877; Dembo, A. et al., (1994) Ann.Pr ob.22:pp. 2022-2039; and Altschul, SF, "Evaluating the statistical significance of multiple distinct local alignments," Theoretical and Computational Methods in Genome Research (ed. S. Suhai), (1997), pp. 1-14, Plenum, NY.

[0059] For example, in the immunoglobulin chains described herein, a “conservatively modified variant” or “conservative substitution” refers to a variant in which one or more amino acids in the polypeptide are substituted by other amino acids having similar characteristics (e.g., charge, side chain size, hydrophobic / hydrophilicity, skeletal conformation and rigidity). Such modifications can be frequently made without significantly interfering with the biological activity of the antibody or fragment. In general, those skilled in the art recognize that a single amino acid substitution in a non-essential region of a polypeptide does not substantially alter its biological activity (e.g., Watson et al., (1987) Molecular Biology). of the Gene, The Benjamin / Cummings Pub.Co., 224 pages (4 th See Ed.). Additionally, substitutions of structurally or functionally similar amino acids are less likely to significantly interfere with biological activity. The present invention includes an anti-IL2Rγ antigen-binding protein containing such a conservatively modified variant immunoglobulin chain.

[0060] Examples of amino acids with side chains having similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Alternatively, conservative substitutions are discussed in Gonnet et al., (1992) Science 256:1443. This refers to any change that has a positive value in the PAM250 log-likelihood matrix disclosed on page 45.

[0061] For example, the anti-IL2Rγ antigen-binding proteins described herein, including variant immunoglobulin chains, may exhibit one or more of the following properties: • Approximately 2.75 × 10 at 25℃ -9 M ~ approx. 3.36×10 -7 M's K D To bind to human IL2Rγ (for example, its fusions such as the myc-myc-His6 fusion); • Approximately 6.42 × 10 at 37℃ -9 M ~ approx. 3.53×10 -7 M's K D And it binds to human IL2Rγ (for example, its fusions such as the myc-myc-His6 fusion). More precisely, approximately 3.53 × 10 -7 K is lower than M. D (to join); • Approximately 3.18 × 10 at 25℃ -9 M ~ approx. 2.38×10 -7 M's K D To bind to cynomolgus monkey IL-2Rγ (for example, its fusions such as the myc-myc-His6 fusion); • Approximately 8.29 × 10 at 37℃ -9 M ~ approx. 3.20×10 -7 M's K DIt binds to cynomolgus monkey IL-2Rγ (for example, its fusions such as the myc-myc-His6 fusion) (or approximately 3.20 × 10⁻¹⁰ -7 K is lower than M. D (to join); • Approximately 2.45 × 10 at 25℃ -9 M ~ approx. 1.20×10 -8 M's K D It binds to human IL2Rγ (for example, its fusion to a C-terminal mouse IgG2a Fc tag, etc.) (or approximately 1.20 × 10⁻¹⁰ -8 K is lower than M. D To join them together); • Approximately 1.86 × 10 at 37℃ -11 M ~ approx. 3.00×10 -8 M's K D It binds to human IL2Rγ (for example, its fusion to a C-terminal mouse IgG2a Fc tag, etc.) (or approximately 3.00 × 10⁻¹⁰ -8 K is lower than M. D To join them together); • Approximately 1.84 × 10 at 25℃ -8 M, 3.76×10 -9 M, 1.08 × 10 -7 M, 2.17×10 -8 M, 6.02×10 -9 M or 7.93 x 10 -8 M's K D To bind (or not bind) to mouse IL2Rγ (for example, its fusion such as the myc-myc-His6 fusion); • Approximately 5.59 × 10 at 37℃ -8 M, 6.11×10 -9 M, 3.87×10 -7 M, 5.16×10 -8 M, 8.70×10 -9 M or 2.15×10 -7 M's K D To bind (or not bind) to mouse IL2Rγ (for example, its fusion such as the myc-myc-His6 fusion); • Approximately 3.32 × 10 at 25℃ -9 M ~ approx. 1.97×10 -7 M's KD Binding (or not binding) to human IL2Rγ domain 1 (for example, its fusions such as the myc-myc-His6 fusion); • Approximately 4.13 × 10 at 37℃ -9 M ~ approx. 2.25×10 -7 M's K D Binding (or not binding) to human IL2Rγ domain 1 (for example, its fusions such as the myc-myc-His6 fusion); • Approximately 2.91 × 10⁻⁶ at 25℃ -7 M ~ approx. 5.35×10 -10 K D Binding (or not binding) to human IL2Rγ domain 2 (for example, its fusions such as the myc-myc-His6 fusion); • Approximately 1.14 × 10 at 37℃ -8 Alternatively, approximately 1.27 × 10 -8 K D Binding (or not binding) to human IL2Rγ domain 2 (for example, its fusions such as the myc-myc-His6 fusion); For example, ICs with a impedance of approximately 1 nM to 0.5 nM 50 For example, T cells (e.g., human CD4) are induced by IL-2 (e.g., approximately 10 nM), IL-4 (e.g., approximately 50 pM), IL-7 (e.g., approximately 1 pM), IL-15 (e.g., approximately 0.5 nM), and / or IL-21 (e.g., approximately 50 pM). + Blocking STAT phosphorylation in T cells; For example, approximately 4 x 10 -10 M IC 50 For example, blocking STAT phosphorylation in mast cells (e.g., differentiated human mast cells) that is induced by IL-9 (e.g., approximately 2 nM); • Mice after being injected with human peripheral blood mononuclear cells (PBMCs) (e.g., NOD-scid To reduce the number of human immune cells (e.g., human PBMCs (peripheral blood mononuclear cells), e.g., human CD45+ cells, human T cells, human CD4+ T cells, and / or human CD8+ T cells) in IL2rγ null (NSG) mice; • Mice after being injected with human peripheral blood mononuclear cells (PBMCs) (e.g., NOD-scid To reduce the levels of serum human cytokines (e.g., human IFN-γ, human TNFα, human IL-6, human IL-8 and / or human IL-10) and / or mouse cytokines (e.g., mouse TNFα, mouse IL-6, mouse KC / GRO and / or mouse IL-10) in IL2rγ null (NSG) mice; • Competing with any one or more anti-IL2Rγ antibodies described herein for binding to human IL-2Rγ (e.g., tagged with a C-terminal myc-myc-hexahistidine tag), for example, on the cell surface; • To bind any one or more anti-IL2Rγ antibodies described herein to the same epitope on IL2Rγ, for example, on the cell surface (e.g., tagged with a C-terminal myc-myc-hexahistidine tag); • Not detectably binding to either mouse IL2Rγ or rat IL2Rγ (for example, by Biacore measurement at 37°C); • To protect mice from weight loss and / or death caused by GvHD in GvHD mouse models; • Blocking the binding of a hybrid receptor containing IL2Rγ complexed with a cytokine-specific receptor subunit to cytokines such as IL-2, IL-4, IL-7, IL-9, IL-15 and / or IL-21; and / or For example, inhibiting intracellular IL2Rγ signaling via the JAK-STAT pathway (e.g., in human B lymphocytes or human natural killer cells) induced by, for example, IL2, IL4, IL7, IL9, IL15 and / or IL21, as measured by luciferase expression in cells containing a luciferase gene operably linked to the STAT3 response element.

[0062] "H4H12857P"; "H4H12858P"; "H4H12859P"; "H4H12863P"; "H4H12874P"; "H4H12871P"; "H4H12884P"; "H4H12886P"; " H4H12889P”; “H4H12890P”; “H4H12899P”; “H4H12900P”; “H4H12908P”; “H4H12913P2”; “H4H12922P2”; “H4H12924P2”; "H4H12926P2"; "H4H12927P2"; "H4H12934P2"; "H4H13538P"; "H4H13541P"; "H4H13544P2"; or "H4H13545P2" are, unless otherwise noted, H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4 For H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2, the amino acid sequences described herein in particular (e.g., SEQ ID NOs: 2, 18, 22, 38, 42, 58, Immunoglobulin heavy chains or their variable regions (V) containing 62, 77, 81, 97, 101, 115, 119, 134, 138, 152, 156, 170, 174, 186, 190, 198, 200, 208, 210, 216, 218, 234, 238, 254, 258, 272, 276, 284, 286, 294, 296, 311, 315, 331, 335, 343, 345, 357, 361 or 376) (or their variants) H), and / or H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H1354 Immunoglobulin light chains or their variable regions (V) containing the amino acid sequences described herein in particular for 1P;H4H13544P2; or H4H13545P2 (e.g., SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 79, 89, 99, 109, 117, 127, 136, 146, 154, 164, 172, 182, 188, 226, 236, 246, 256, 266, 274, 304, 313, 323, 333, 353, 359, 368 or 378) (or their variants) L ) each include; and / or a heavy chain or V containing the CDR (CDR-H1 (or its variant), CDR-H2 (or its variant), and CDR-H3 (or its variant)) H and / or light chain or V containing the CDR (CDR-L1 (or its variant), CDR-L2 (or its variant), and CDR-L3 (or its variant)) L This refers to anti-IL2Rγ antigen-binding proteins, such as antibodies and their antigen-binding fragments (including multispecific antigen-binding proteins). In embodiments of the present invention, V H It is linked to the IgG constant heavy chain domain, for example, the human IgG constant heavy chain domain (e.g., IgG1 or IgG4 (e.g., including S228P and / or S108P mutations)), and / or V L It is linked to a light chain constant domain, for example, a human light chain constant domain (e.g., a lambda or kappa constant light chain domain). Any such immunoglobulin chain (e.g., V H , V LA polynucleotide encoding one or more of HC and / or LC forms a portion of the present invention.

[0063] The present invention includes a “neutralizing” or “antagonist” anti-IL2Rγ antigen-binding protein (e.g., an antibody or antigen-binding fragment) that contains a molecule that inhibits the activity of IL2Rγ (e.g., the binding of a hybrid receptor containing IL2Rγ complexed with a cytokine-specific receptor subunit to cytokines such as IL-2, IL-4, IL-7, IL-9, IL-15, and / or IL-21) to any detectable extent.

[0064] The antibody and antigen-binding fragment of the present invention (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P; H4H129 13P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2) comprises an immunoglobulin chain including cellular and in vitro post-translational modifications to an antibody or fragment, in addition to the amino acid sequences (and their variants) described herein in particular. For example, the present invention includes antibodies and their antigen-binding fragments that specifically bind to IL2Rγ containing the heavy-chain and / or light-chain amino acid sequences described herein, in addition to antibodies and fragments in which one or more asparagine, serine and / or threonine residues are glycosylated, one or more asparagine residues are deamidated, one or more residues (e.g., Met, Trp and / or His) are oxidized, the N-terminal glutamine is pyroglutamic acid (pyroE), and / or a C-terminal lysine or other amino acid is deleted.

[0065] The present invention relates to anti-IL2Rγ antigen-binding proteins, such as H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4 The present invention provides containers containing H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2; or H4H13545P2 (for example, plastic or glass vials having caps or chromatography columns, hollow needles or syringe cylinders).

[0066] The present invention also relates to one or more antigen-binding proteins (e.g., antibodies or antigen-binding fragments) that specifically bind to IL2Rγ, e.g., H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12 The present invention provides an injection device containing 890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2, or a pharmaceutical formulation thereof. The injection device can be packaged in a kit. The injection device is a device for introducing a substance into a target body via parenteral routes, such as intraocular, intravitreous, intramuscular, subcutaneous, or intravenous routes. For example, the injection device may be a syringe or autoinjector (e.g., pre-filled with the pharmaceutical formulation) that includes, for example, a cylinder or barrel for holding the fluid to be injected (e.g., an antibody or fragment or its pharmaceutical formulation), a needle for piercing the skin, blood vessel or other tissue for the injection of the fluid, and a plunger for pushing the fluid out of the cylinder through the needle hole into the target body.

[0067] The present invention further relates to the anti-IL2Rγ antigen-binding proteins of the present invention, for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H1291 The present invention provides a method for administering 3P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2; or H4H13545P2 to a target, comprising introducing an antigen-binding protein into the body of a target (e.g., a human), for example, parenterally. For example, the method comprises inserting a syringe needle into the body of the target and injecting the antigen-binding protein into the body of the target, for example, into the target's vein, artery, eye, muscle tissue, or subcutaneous tissue.

[0068] Polynucleotides and methods for their production Examples of polynucleotides include DNA and RNA. The present invention includes, for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P; Immunoglobulin V of the form H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 H , V L The present invention comprises any polynucleotide of the present invention that encodes CDR-H, CDR-L, HC, or LC, and optionally operably ligated to a promoter or other expression control sequence. For example, the present invention includes SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 74, 76, 78, 80, 82, 84, 86, 88, 90, 9 2, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237 ,239,241,243,245,247,249,251,253,255,257,259,261,263,265,267,269,271,273,275,277,279,281,283,285,287,289,291,293,295,297,299,301,303,305,307,308,310 The present invention provides any polynucleotide (e.g., DNA) comprising the nucleotide sequences described in 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 365, 367, 369, 371, 373, 375, or 377. In embodiments of the present invention, the polynucleotide of the present invention is fused to a secretion signal sequence. A polypeptide encoded by such a polynucleotide is also within the scope of the present invention.

[0069] Generally, a “promoter” or “promoter sequence” is a DNA regulatory region that can bind to RNA polymerase in a cell (for example, directly or through a protein or substance bound to another promoter) and initiate transcription of a coding sequence. Promoters may be operably ligated to other expression regulatory sequences, including enhancer and repressor sequences, and / or to the polynucleotides of the present invention. Promoters that can be used to control gene expression include the cytomegalovirus (CMV) promoter (U.S. Patent Nos. 5,385,839 and 5,168,062) and the SV40 initial promoter region (Benoist et al., (1981)N ature 290:304~310pp), promoter contained in the 3' long chain terminal repeat of Rous sarcoma virus (Yamamoto et al., (1980) Cell 22:787~ (p. 797), herpesthymidine kinase promoter (Wagner et al., (1981)P (roc.Natl.Acad.Sci.USA 78:1441~1445), regulatory sequence of the metallothionein gene (Brinster et al., (1982) Nature 296) pp. 39-42); Prokaryotic expression vectors, e.g., beta-lactamase promoter (VIIIa-Komaroff et al., (1978) Proc. Natl. Acad. Sci. USA 75:3727-3731), or tac promoter (DeBoer et al., (1983) Proc. Natl. Acad. Sci. USA 80:21-25); "Useful proteins from recombinant bacteria", Scientific American (1980) 242:74-94. See also promoter elements from yeast or other fungi, such as the Gal4 promoter, ADC (alcohol dehydrogenase) promoter, PGK (phosphoglycerol kinase) promoter, or alkaline phosphatase promoter, but not limited to these.

[0070] In cells or other expression systems, a promoter or other regulatory sequence directs RNA polymerase-mediated transcription of a coding sequence to RNA, preferably mRNA, and optionally the RNA, preferably mRNA, is then RNA spliced ​​(if it contains introns) and optionally translated into a protein encoded by the coding sequence, in which case the polynucleotide encoding the polypeptide is "operably ligated" to the promoter or other regulatory sequence.

[0071] Invention 5V H and V L Includes polynucleotides containing the following polynucleotide pairs that encode: Sequence IDs 1 and 9; Sequence IDs 21 and 29; Sequence IDs 41 and 49; Sequence IDs 61 and 69; Sequence IDs 80 and 88; Sequence IDs 100 and 108; Sequence IDs 118 and 126; Sequence IDs 137 and 145; Sequence IDs 155 and 163; Sequence IDs 173 and 181; Sequence IDs 189 and 181; Sequence IDs 199 and 181; Sequence IDs 209 and 181; Sequence IDs 217 and 225; Sequence IDs 237 and 245; Sequence IDs 257 and 265; Sequence IDs 275 and 181; Sequence IDs 285 and 181; Sequence IDs 295 and 303; Sequence IDs 314 and 322; Sequence IDs 334 and 181; Sequence IDs 344 and 352; or Sequence IDs 360 and 367.

[0072] The present invention comprises a polynucleotide comprising the following set of polynucleotides encoding CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3: Sequence IDs 3, 5, 7, 11, 13, and 15; Sequence IDs 23, 25, 27, 31, 33, and 35; Sequence IDs 43, 45, 47, 51, 53, and 55; Sequence IDs 63, 65, 67, 71, 73, and 74; Sequence IDs 82, 84, 86, 90, 92, and 94; Sequence IDs 102, 104, 106, 110, 73, and 112; Sequence IDs 120, 122, 124, 128, 130, and 131; Sequence IDs 139, 141, 143, 147, 73, and 149; Sequence IDs 157, 159, 161, 165, 13, and 167; Sequence IDs 175, 177, 179, 71, 73, and 183; Sequence IDs 191, 193, 195, 71, 73, and 183; Sequence IDs 201, 203, 205, 71, 73, and 183; Sequence IDs 175, 211, 213, 71, 73, and 183; Sequence IDs 219, 221, 223, 227, 229, and 231; Sequence IDs 239, 241, 243, 247, 249, and 251; Sequence IDs 259, 261, 263, 267, 73, and 269; Sequence IDs 277, 279, 281, 71, 73, and 183; Sequence IDs 287, 289, 291, 71, 73, and 183; Sequence IDs 297, 299, 301, 305, 307, and 308; Sequence IDs 316, 318, 320, 324, 326, and 328; Sequence IDs 336, 338, 340, 71, 73, and 183; Sequence IDs 346, 348, 350, 71, 73 and 354; or Sequence numbers 362, 364, 365, 369, 371, and 373.

[0073] The present invention includes a polynucleotide comprising the following polynucleotide pairs encoding HC and LC: Sequence IDs 17 and 19; Sequence IDs 37 and 39; Sequence IDs 57 and 59; Sequence IDs 76 and 78; Sequence IDs 96 and 98; Sequence IDs 114 and 116; Sequence IDs 133 and 135; Sequence IDs 151 and 153; Sequence IDs 169 and 171; Sequence IDs 185 and 187; Sequence IDs 197 and 187; Sequence IDs 207 and 187; Sequence IDs 215 and 187; Sequence IDs 233 and 235; Sequence IDs 253 and 255; Sequence IDs 271 and 273; Sequence IDs 283 and 187; Sequence IDs 293 and 187; Sequence IDs 310 and 312; Sequence IDs 330 and 332; Sequence IDs 342 and 187; Sequence IDs 356 and 358; or Sequence IDs 375 and 377.

[0074] The present invention includes polynucleotides encoding immunoglobulin polypeptide chains whose nucleotide sequences are variants of polynucleotides specifically described herein. A “variant” of a polynucleotide refers to a polynucleotide that contains a nucleotide sequence identical to at least about 70–99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) of the reference nucleotide sequences described herein, when the comparison is performed by the BLAST algorithm and the parameters of the algorithm are selected to give the maximum match between each sequence over the full length of each reference sequence (e.g., expected threshold: 10; word size: 28; maximum match within query range: 0; match / mismatch score: 1, -2; gap cost: linear). In embodiments of the present invention, variants of nucleotide sequences, as specifically described herein, include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) point mutations, insertions (e.g., in-frame insertions), or deletions (e.g., in-frame deletions) of one or more nucleotides. Such mutations may be missense or nonsense mutations in embodiments of the present invention. In embodiments of the present invention, such variant polynucleotides can be incorporated into an anti-IL2Rγ antigen-binding protein, i.e., encoding an immunoglobulin polypeptide chain such that the protein maintains specific binding to IL2Rγ.

[0075] Eukaryotic and prokaryotic host cells, including mammalian cells, can be used as hosts for the expression of anti-IL2Rγ antigen-binding proteins (e.g., antibodies or their antigen-binding fragments). Such host cells are well known in the art and many are available from the American Type Culture Collection (ATCC). These host cells include, among others, Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells, and numerous other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cattle, horse, and hamster cells. Other cell lines that can be used include insect cell lines (e.g., Spodoptera frugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plant cells, and fungal cells. Fungal cells include yeast and filamentous fungal cells, which include, for example, *P. Pichia, Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijuperi Pichia pijperi, Pichia stiptis, Pichia metanolica methanolica), Pichia species, Saccharomyces cerevisiae, Saccharomyces species, Hansenula polymorpha, Kluyveromyces species, Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger This invention includes niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium species, Fusarium gramineum, Fusarium venenatum, Physcomitrella patens, and Neurospora crassa. The present invention relates to antigen-binding proteins, their V H , V L HC, LC, or CDR (or their variants), for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924 The isolated host cell (e.g., a CHO cell or any of the host cells described above) comprises P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2; and / or one or more polynucleotides encoding its immunoglobulin chain (e.g., as discussed herein).

[0076] The present invention also relates to antigen-binding proteins (e.g., antibodies or their antigen-binding fragments), e.g., H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2; The present invention comprises cells expressing IL2Rγ or its antigenic fragments or fusions (e.g., His6, Fc and / or myc) conjugated by H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2; or H4H13545P2, where the cells are, for example, present in the body of the subject or in vitro.

[0077] In addition, the present invention also relates to the present invention in complex with an IL2Rγ polypeptide or its antigenic fragment or fusion thereof and / or an anti-IL2Rγ antibody or a secondary antibody or its antigen-binding fragment that specifically binds to the fragment (e.g., a detectably labeled secondary antibody). The present invention provides a complex comprising an anti-IL2Rγ antigen-binding protein, such as an antibody or its antigen-binding fragment, as discussed in the book. In embodiments of the present invention, the complex is in vitro (e.g., immobilized on a solid substrate) or present in the body of a subject.

[0078] The recombinant anti-IL2Rγ antigen-binding proteins disclosed herein, e.g., antibodies and antigen-binding fragments, can also be produced in an E. coli / T7 expression system. In this embodiment, the anti-IL2Rγ antibody immunoglobulin molecules of the present invention (e.g., H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H128 HC, LC, V of 99P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 H and / or V LA polynucleotide encoding either or its CDR can be inserted into a pET-based plasmid and expressed in the E. coli / T7 system. For example, the present invention provides a method for expressing an antibody or its antigen-binding fragment or its immunoglobulin chain in a host cell (e.g., a bacterial host cell, such as E. coli BL21 or BL21DE3), wherein the polynucleotide encoding an immunoglobulin chain operably linked to a T7 promoter (e.g., SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 175, 177, 179 ,181,183,185,187,189,191,193,195,197,199,201,203,205,207,209,211,213,215,217,219,221,223,225,227,229,231,233,235,237,239,241,243,245,247,249,251,253,255,257,259,261,263,265,267,269,271,273,275,277,279,281,283,285,287,289,291,29 A method comprising expressing T7 RNA polymerase in cells that also include a nucleotide sequence in any one or more of 3, 295, 297, 299, 301, 303, 305, 307, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 365, 367, 369, 371, 373, 375, or 377; or a variant thereof.For example, in embodiments of the present invention, a bacterial host cell such as E. coli contains a polynucleotide encoding a T7 RNA polymerase gene operably linked to a lac promoter, and the expression of the polymerase and chain is induced by incubation of the host cell with IPTG (isopropyl-beta-D-thiogalactopyranoside). See US4952496 and US5693489 or Studier & Moffatt, "Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes," J.Mol.Biol.1986 May 5;189(1):113-130.

[0079] There are several methods known in the art for producing recombinant antibodies. One example of a method for recombinant antibody production is disclosed in US4816567.

[0080] Transformation can be carried out by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, polynucleotide encapsulation in liposomes, microparticle gun injection of DNA into the nucleus, and direct microinjection. In addition, nucleic acid molecules can be introduced into mammalian cells by viral vectors. Methods for transforming cells are well known in the art. See, for example, U.S. Patent No. 4,399,216; No. 4,912,040; No. 4,740,461 and No. 4,959,455.Therefore, the present invention relates to a recombinant method for producing an anti-IL2Rγ antigen-binding protein, for example, the antibody of the present invention or its antigen-binding fragment, or its immunoglobulin chain, wherein (i) an antigen-binding protein, for example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922 P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2;one or more polynucleotides encoding the immunoglobulin light and / or heavy chains (e.g., SEQ ID NOs: 1, 9, 17, 19, 21, 29, 37, 39, 41, 49, 57, 59, 61, 69, 76, 78, 80, 88, 96, 98, 100, 108, 114, 116, 118, 126, 133, 135, 137, 145, 151) , nucleotide sequences in one or more of the following locations: 153, 155, 163, 169, 171, 173, 181, 185, 187, 189, 197, 199, 207, 209, 215, 217, 225, 233, 235, 237, 245, 253, 255, 257, 265, 271, 273, 275, 283, 285, 293, 295, 303, 310, 312, 314, 322, 330, 332, 334, 342, 344, 352, 356, 358, 360, 367, 375 or 377; including variants thereof) in the host The method includes (ii) introducing into cells, for example, a polynucleotide in a vector; and / or integrated into a host cell chromosome and / or operably ligated to a promoter; (ii) culturing host cells (e.g., CHO or Pichia or Pichia pastris) under conditions favorable to the expression of the polynucleotide; and (iii) optionally isolating an antigen-binding protein (e.g., an antibody or antigen-binding fragment) or chain from the host cells and / or the culture medium in which the host cells were grown.When producing antigen-binding proteins (e.g., antibodies or antigen-binding fragments) containing more than one immunoglobulin chains, for example, an antibody containing two immunoglobulin heavy chains and two immunoglobulin light chains, the co-expression of the chains in a single host cell leads to the association of the chains, for example, within or on the cell surface, or outside the cell if such chains are secreted, forming an antigen-binding protein (e.g., an antibody or antigen-binding fragment). The methods of the present invention include methods in which only immunoglobulin heavy chains, or only immunoglobulin light chains, or both (e.g., any mature fragment and / or its variable domain as discussed herein) are expressed in cells. Such a single chain is useful, for example, as an intermediate in the expression of an antibody or antigen-binding fragment containing such a chain. For example, the present invention also includes anti-IL2Rγ antigen-binding proteins, for example, antibodies and their antigen-binding fragments, which are products of the production methods and optionally the purification methods described herein.

[0081] In embodiments of the present invention, a method for producing an anti-IL2Rγ antigen-binding protein, such as an antibody or its antigen-binding fragment, includes a method for purifying the antigen-binding protein, for example, by column chromatography, precipitation, and / or filtration. As will be discussed, the products of such methods also form part of the present invention.

[0082] Manufacturing of human antibodies The anti-IL2Rγ antibody of the present invention may be a fully human antibody. Methods for producing monoclonal antibodies, including fully human monoclonal antibodies, are known in the art. Any such known method can be used in the context of the present invention to produce a human antibody that specifically binds to human IL2Rγ.

[0083] For example, when using VELOCIMMUNE® technology or any other similar known method for generating fully human monoclonal antibodies, a high-affinity chimeric antibody against IL2Rγ having a human variable region and a mouse constant region is first isolated. As in the experimental section below, the antibody is characterized and selected for desirable features, including affinity, ligand blocking activity, selectivity, and epitope. If necessary, to generate a fully human anti-IL2Rγ antibody, the mouse constant region is replaced with a desired human constant region, e.g., wild-type or modified IgG1 or IgG4. The selected constant region may vary depending on the specific application, but the high-affinity antigen-binding and target specificity features reside in the variable region. In certain cases, fully human anti-IL2Rγ antibodies are isolated directly from antigen-positive B cells. See, for example, US 6,596,541, Regeneron Pharmaceuticals, VELOCIMMUNE®.

[0084] Anti-IL2Rγ antibody containing Fc variant According to a particular embodiment of the present invention, for example, an anti-IL2Rγ antibody is provided comprising an Fc domain containing one or more mutations that enhance or decrease antibody binding to the FcRn receptor at acidic pH compared to neutral pH. For example, the present invention provides an Fc domain containing C H 2 or C H The present invention relates to an anti-IL2Rγ antibody containing mutations in three regions, wherein the mutations increase the affinity of the Fc domain to FcRn in an acidic environment (e.g., in endosomes where the pH is in the range of approximately 5.5 to 6.0). Such mutations may result in an increase in the serum half-life of the antibody when administered to animals.

[0085] An example of such a non-limiting Fc modification is, for instance, position: 250 (for example, E or Q); 250 and 428 (for example, L or F); 252 (for example, L / Y / F / W or T), · 254 (for example, S or T), and / or 256 (for example, S / R / Q / E / D or T) Modifications in; and / or location: · 428 and / or 433 (e.g., H / L / R / S / P / Q or K), and / or ·434 (for example, H / F or Y) Modifications in; and / or location: 250 and / or 428 Modifications in; and / or location: 307 or 308 (e.g., 308F, V308F), and / or 434 Modifications in These are some examples.

[0086] In embodiments of the present invention, modifications include: Modifications of 428L (e.g., M428L) and 434S (e.g., N434S); Modifications of 428L, 259I (e.g., V259I), and 308F (e.g., V308F); Modifications of 433K (e.g., H433K) and 434 (e.g., 434Y); Modifications of 252, 254, and 256 (e.g., 252Y, 254T, and 256E); • Modifications of the 250Q and 428L (e.g., T250Q and M428L); and / or Modifications of 307 and / or 308 (e.g., 308F or 308P) Includes.

[0087] For example, the present invention: • 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T, and 256E (e.g., M252Y, S254T, and T256E); · 428L and 434S (e.g., M428L and N434S); and · 433K and 434F (e.g., H433K and N434F) An anti-IL2Rγ antibody comprising an Fc domain that includes one or more pairs or groups of mutations selected from the group consisting of

[0088] In an embodiment of the invention, the heavy chain constant domain is γ4 comprising the S228P and / or S108P mutations. See Angal et al., “A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody” , Mol Immunol. 1993 Jan; 30(1): 105-108.

[0089] ​​​​​​​​In certain embodiments of the present invention, the modified Fc domain is a variant IgG1 Fc or variant IgG4 Fc that includes substitutions in the hinge region. For example, the modified Fc for use in the context of the present invention may include a variant IgG1 Fc in which at least one amino acid of the IgG1 Fc hinge region is replaced with the corresponding amino acid from the IgG2 Fc hinge region. Alternatively, the modified Fc for use in the context of the present invention may include a variant IgG4 Fc in which at least one amino acid of the IgG4 Fc hinge region is replaced with the corresponding amino acid from the IgG2 Fc hinge region. Non-limiting exemplary modified Fc regions that can be used in the context of the present invention include, in addition to those described in U.S. Patent Application Publication No. 2014 / 0243504, which is hereby incorporated by reference in its entirety, any functionally equivalent variant of the modified Fc regions described therein. are any functionally equivalent variant of the modified Fc regions described therein.

[0092] Other modified Fc domains and Fc modifications that can be used in the context of the present invention include any modifications as described in US2014 / 0171623; US8697396; US2014 / 0134162; WO2014 / 043361, which are hereby incorporated by reference in their entirety. Methods of constructing antibodies or other antigen-binding fusion proteins that include a modified Fc domain as described herein are known in the art.

[0093] Multispecific antigen-binding protein The present invention includes anti-IL2Rγ antigen-binding proteins, such as antibodies and their antigen-binding fragments, as well as methods for using them and methods for producing such antigen-binding proteins. The terms "anti-IL2Rγ" or "anti-IL2R gamma" antigen-binding protein, such as antibodies or antigen-binding fragments, refer to at least one first antigen-binding domain that specifically binds to IL2Rγ (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P; The present invention comprises a multispecific (e.g., bispecific or biparatope) molecule comprising an antigen-binding domain from H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2) and at least one second antigen-binding domain that binds to a different antigen or an epitope in a different IL2Rγ than the first antigen-binding domain. In embodiments of the present invention, the first and second epitopes overlap. In another embodiment of the present invention, the first and second epitopes do not overlap.

[0094] Multiple specificity refers to binding to two or more different epitopes, which may be located on the same or different antigens. Examples of multiple specificity include bispecificity, tripspecificity, and quadrupspecificity.

[0095] "H4H12857P"; "H4H12858P"; "H4H12859P"; "H4H12863P"; "H4H12874P"; "H4H12871P"; "H 4H12884P”; “H4H12886P”; “H4H12889P”; “H4H12890P”; “H4H12899P”; “H4H12900P”; “H4H "12908P"; "H4H12913P2"; "H4H12922P2"; "H4H12924P2"; "H4H12926P2"; "H4H12927P2"; "H4H12934P2"; "H4H13538P"; "H4H13541P"; "H4H13544P2"; or "H4H13545P2" are all "H4H" 12857P"; "H4H12858P"; "H4H12859P"; "H4H12863P"; "H4H12874P"; "H4H12871P"; "H4H12 884P";"H4H12886P";"H4H12889P";"H4H12890P";"H4H12899P";"H4H12900P";"H4H1290 HCDR and LCDR of "8P"; "H4H12913P2"; "H4H12922P2"; "H4H12924P2"; "H4H12926P2"; "H4H12927P2"; "H4H12934P2"; "H4H13538P"; "H4H13541P"; "H4H13544P2"; or "H4H13545P2", V H and V L , or HC and LC, as well as multispecific molecules, such as antibodies or antigen-binding fragments, that include one or more antigen-binding domains that bind to different epitopes.

[0096] In embodiments of the present invention, the antigen-binding domain that specifically binds to IL2Rγ and can be included in the multispecific molecule is: (1) (i) Heavy chain variable domains (V) containing CDR-H1, CDR-H2 and CDR-H3 from an immunoglobulin heavy chain containing an amino acid sequence (or a variant thereof) selected from SEQ ID NOs: 2, 22, 42, 62, 81, 101, 119, 138, 156, 174, 190, 200, 210, 218, 238, 258, 276, 286, 296, 315, 335, 345 and 361 H) array, and (ii) Light chain variable domains (V) containing CDR-L1, CDR-L2 and CDR-L3 from an immunoglobulin light chain containing an amino acid sequence (or a variant thereof) selected from SEQ ID NOs: 10, 30, 50, 70, 89, 109, 127, 146, 164, 182, 226, 246, 266, 304, 323, 353 and 368 L )array; or (2) (i) Heavy chain variable domain (V) containing an amino acid sequence (or a variant thereof) selected from SEQ ID NOs: 2, 22, 42, 62, 81, 101, 119, 138, 156, 174, 190, 200, 210, 218, 238, 258, 276, 286, 296, 315, 335, 345 and 361 H ); and (ii) Light chain variable domain (V) containing an amino acid sequence (or a variant thereof) selected from SEQ ID NOs: 10, 30, 50, 70, 89, 109, 127, 146, 164, 182, 226, 246, 266, 304, 323, 353 and 368 L ); Furthermore One or more antigen-binding domains that bind to different epitopes Includes.

[0097] In one embodiment of the present invention, the bispecific antigen-binding fragment is a first scFv (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12 V of 889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 H and V LThe first and second scFv include a linker, e.g., a peptide linker (e.g., a GS linker, e.g., (GGGGS)) and a second scFv having binding specificity to a different epitope. For example, in embodiments of the present invention, the first and second scFv include a linker, e.g., a peptide linker (e.g., a GS linker, e.g., (GGGGS)) n (Sequence number 386) (where n is, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10)) is used to connect them.

[0098] Other bispecific antigen-binding fragments include H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2 Examples include H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 and F(ab)2, a bispecific IgG antibody containing heavy and light chain CDRs of another antibody that binds to a different epitope.

[0099] Immunoconjugate The present invention also includes another part, for example, an anti-IL2Rγ antigen-binding protein, for example, an antibody or antigen-binding fragment ("immunoconjugate") conjugated to a therapeutic part. In embodiments of the present invention, the anti-IL2Rγ antigen-binding protein, for example, an antibody or antigen-binding fragment is the It is conjugated to any further therapeutic agent described in the specification. As used herein, the term “immunoconjugate” refers to an antigen-binding protein, such as an antibody or antigen-binding fragment, chemically or biologically linked to another antigen-binding protein, drug, radioactive agent, reporter moiety, enzyme, peptide, protein, or therapeutic agent.

[0100] Administration and treatment The present invention relates to a method for treating or preventing an IL2Rγ-mediated disease or condition in a subject, comprising a therapeutically effective dose of anti-IL2Rg antigen-binding protein (H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890 The present invention provides a method comprising administering P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2) to a target.

[0101] "IL2Rγ-mediated disease or condition" is any disease condition whose symptoms are mediated by the activity of one or more cytokines IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 and / or receptors that bind to such cytokines; for example, autoimmune and / or inflammatory conditions mediated by such cytokines and / or receptors. For example, IL2Rγ-mediated diseases or conditions include graft-versus-host disease (GvHD), organ graft rejection (e.g., skin grafts, β-islet cell grafts, heart grafts, lung grafts, kidney grafts and / or liver grafts), shotgun chorioretinopathy, multiple sclerosis, uveitis, autoimmune diseases (e.g., type 1 diabetes mellitus, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, and myasthenia gravis), aplastic anemia; atopic dermatitis; asthma; and mast cell activation disorders (e.g., mast cell activation syndrome (MCAS), systemic hypertrophic cell disease (SM), or mast cell leukemia (MCL)).

[0102] The present invention also relates to antigen-binding proteins (e.g., antibodies or their antigen-binding fragments) that specifically bind to IL2Rγ, e.g., H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H A method for administering 12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 to a subject having, for example, an IL2Rγ-mediated disease or condition, comprising introducing the antigen-binding protein into the subject's body, for example, by injection.

[0103] Graft-versus-host disease (GvHD) is a condition that can occur after allogeneic transplantation. For example, in GvHD, donated bone marrow or peripheral blood stem cells may perceive the recipient's body as foreign, and the donated cells / bone marrow may attack the body. GvHD can occur, for example, after hematopoietic stem cell transplantation (HCT; for example, in subjects with acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL) and / or myelodysplastic syndrome or myeloproliferative neoplasm), blood transfusion, thym transplantation, or in patients with thymoma. Types of GvHD include steroid-refractory GvHD, acute graft-versus-host disease (aGvHD), and chronic graft-versus-host disease (cGvHD). Allogeneic transplant recipients may experience either aGvHD or cGvHD, or both, or neither. The present invention relates to a method for treating or preventing GvHD (of any type) in a subject. The present invention includes a method comprising administering a therapeutically effective dose of anti-IL2Rγ antigen-binding protein to a target.

[0104] Symptoms of aGvHD may include skin rash or reddened areas on the skin (signs of aGvHD on the skin); yellowing of the skin and / or eyes, and abnormal blood test results (signs of aGvHD on the liver); nausea, vomiting, diarrhea, or abdominal cramps (signs of aGvHD in the gastrointestinal tract, or "intestines"); and / or increased dryness / irritation of the eyes (signs of GvHD on the eyes).

[0105] Symptoms of cGvHD may include rash, raised or discolored areas, thickening or hardening of the skin (signs of cutaneous cGvHD); abdominal distension, yellowing of the skin and / or eyes, and abnormal blood test results (signs of hepatic cGvHD); dry eyes or visual changes (signs of ocular cGvHD); dry mouth, white spots in the mouth, pain or hypersensitivity to spicy foods (signs of oral cGvHD); shortness of breath or changes seen on chest X-ray (signs of pulmonary cough pulmonary cGvHD); difficulty swallowing, pain with swallowing, or weight loss (signs of gastrointestinal tract or "intestine" cGvHD); fatigue, muscle weakness, or pain (signs of neuromuscular cGvHD); and / or increased need to urinate (frequency of urination), burning or bleeding with urination, vaginal dryness / hardening, or penile dysfunction (signs of genitourinary system, bladder, or reproductive organs cGvHD).

[0106] Organ transplant rejection is the rejection of a transplanted organ by the recipient's immune system. Hyperacute rejection occurs within minutes of transplantation, acute rejection occurs within one week to three months after transplantation, and chronic rejection occurs over many years. Examples of organs that can be transplanted include solid organs such as skin, pancreas, kidney, liver, heart, and lungs. The present invention includes a method for treating or preventing organ transplant rejection (of any kind) in a subject, comprising administering a therapeutically effective dose of anti-IL2Rγ antigen-binding protein to the subject.

[0107] Scatter chorioretinopathy is a rare form of posterior uveitis, i.e., inflammation of the uvea, the part of the eye that provides most of its blood supply to the retina. Scatter chorioretinopathy can be caused by autoimmunity. Symptoms of scatter chorioretinopathy may include night blindness, color vision problems, sensitivity to bright light, seeing flashing lights, visual distortion, pain in the eye, and loss of depth perception and / or peripheral vision. The present invention provides a method for treating or preventing scatter chorioretinopathy or uveitis in a subject, comprising, for example, administering a therapeutically effective dose of anti-IL2Rγ antigen-binding protein to the subject by intraocular administration, e.g., intravitreal injection.

[0108] The present invention also provides a method for treating or preventing any autoimmune disease or condition by inhibiting IL2Rγ. Blocking the signaling of one or more cytokines of the γc family may be beneficial in patients suffering from autoimmunity due to their inhibitory effects on the secretion of inflammatory cytokines and the production of autoantibodies. Multiple sclerosis (MS) is a disease of the brain and spinal cord (central nervous system (CNS)) in which the immune system attacks the myelin sheath of nerve fibers, causing problems with communication between the brain and other parts of the body. Ultimately, the disease can cause deterioration or permanent damage to the nerves themselves. Rheumatoid arthritis (RA) is an autoimmune disease in which the body's immune system attacks the joints. This causes thickening of the tissue (synovial membrane) that lines the inside of the joints, generating inflammation that results in swelling and pain in and around the joints. Psoriasis is an autoimmune disease with a primary symptom affecting the skin. Inflammation can also affect the joints, vascular system, and eyes of people with psoriasis. Type 1 diabetes is an autoimmune disease in which the immune system attacks and destroys insulin-producing beta cells in the pancreas. The pancreas then produces little to no insulin. Systemic lupus erythematosus (SLE) is an autoimmune disease in which the body's immune system attacks its own tissues and organs. It is a systemic autoimmune disease that occurs in combination. The inflammation caused by lupus can affect many different body systems, including joints, skin, kidneys, blood cells, brain, heart and lungs. Myasthenia gravis is an autoimmune disease in which antibodies block the acetylcholine receptor at the neuromuscular junction, preventing muscle contraction. In most individuals with myasthenia gravis, this is caused by antibodies against the acetylcholine receptor itself. However, antibodies against other proteins, such as the MuSK (muscle-specific kinase) protein, can also lead to impaired transmission at the neuromuscular junction. The present invention includes a method of treating or preventing an autoimmune disorder or condition (e.g., multiple sclerosis or any other central nervous system inflammation, rheumatoid arthritis, psoriasis, type I diabetes, systemic lupus erythematosus and / or myasthenia gravis) in a subject, the method comprising administering to the subject a therapeutically effective dosage of an anti-IL2Rγ antigen-binding protein.

[0109] The effective or therapeutically effective dose of an anti-IL2Rg antigen-binding protein, such as an antibody or antigen-binding fragment, for treating or preventing an IL2Rγ-mediated disease or condition refers to an amount of antigen-binding protein sufficient to alleviate one or more signs and / or symptoms of the disease or condition in the subject being treated, whether by inducing regression or elimination of such signs and / or symptoms, or by inhibiting the progression of such signs and / or symptoms. In embodiments of the present invention, the effective or therapeutically effective dose of the anti-IL2Rg antigen-binding protein is approximately 0.05 to 50 mg per kg of body weight. The dose may vary depending on the age and size of the subject being administered to, the target disease, condition, and route of administration, etc. In certain embodiments, an initial dose may be followed by a second or more subsequent doses of antigen-binding protein in amounts that are approximately the same as, less than, or more than, the initial dose, with intervals of at least 1 to 3 days; at least 1 week; at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks.

[0110] As used herein, the term “Subject” means, for example, a mammal (e.g., rat, mouse, cat, dog, cattle, sheep, horse, goat, rabbit), preferably a human, that requires the prevention and / or treatment of an IL2Rγ-mediated disease. The subject may have an IL2Rγ-mediated disease or be predisposed to developing such a disease.

[0111] "Preventing" an IL2Rγ-mediated disease or condition, in the context of the use of the anti-IL2Rγ antigen-binding protein of the present invention, means administering it to a subject in such a way that such an occurrence does not occur before the onset of the disease or condition in the subject's body.

[0112] Combinations and pharmaceutical formulations The present invention provides not only compositions comprising an anti-IL2Rγ antigen-binding protein together with one or more components, but also methods for using such compositions and methods for preparing such compositions. Pharmaceutical formulations comprising an anti-IL2Rγ antigen-binding protein and a pharmaceutically acceptable carrier or excipient are part of the present invention.

[0113] Anti-IL2Rγ antigen-binding proteins, for example, antibodies and their antigen-binding fragments (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H To manufacture pharmaceutical formulations of 4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2), antigen-binding proteins are pharmaceutically acceptable carriers or excipients. It is mixed with a modifier. For example, Remington's Pharmaceutical Sciences and US Pharmacopeia: National Formulary, Mack Publishing Company, Easton, Pa. (1984); Hardman et al., (2001) Goodman and Gil man's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practic e of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis et al. (eds.) (1993) Pharma Ceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Mar cel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmac eutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) See Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY. In embodiments of the present invention, the pharmaceutical formulation is sterile. Such a composition is part of the present invention.

[0114] The pharmaceutical formulation of the present invention comprises an anti-IL2Rγ antigen-binding protein, as well as a pharmaceutically acceptable carrier, for example, water, a buffering agent, a preservative, and / or a surfactant.

[0115] The scope of the present invention is limited to anti-IL2Rγ antigen-binding proteins, such as antibodies or their antigen-binding fragments (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P) This includes a dried, for example, freeze-dried composition containing ;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2), or a pharmaceutical formulation thereof containing a pharmaceutically acceptable carrier but substantially lacking water.

[0116] Further embodiments of the present invention disclose anti-IL2Rγ antigen-binding proteins, such as antibodies or antigen-binding fragments thereof (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; Further therapeutic agents administered to the subject along with H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2) are as described in the Physicians' Desk Reference 2003. (Thomson Healthcare; 57) th The medication will be administered to the subjects according to the edition (Nov. 1, 2002) of the edition.

[0117] The mode of administration of anti-IL2Rγ antigen-binding proteins or compositions thereof can vary. Routes of administration include parenteral, non-parenteral, oral, rectal, transmucosal, intestinal; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, inhalation, blown, topical, cutaneous, intraocular, intravitreous, percutaneous, or intraarterial.

[0118] The present invention relates to an anti-IL2Rγ antigen-binding protein, for example, an antibody or its antigen-binding fragment ( For example, H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H1 The present invention provides a method for administering a protein or a pharmaceutical formulation thereof to a subject (2922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2), comprising introducing a protein or a pharmaceutical formulation thereof into the subject's body. For example, in embodiments of the present invention, the method comprises, for example, inserting a syringe needle into the subject's body and injecting the antigen-binding protein or a pharmaceutical formulation thereof into the subject's body, for example, the subject's eye, vein, artery, muscle tissue or subcutaneous tissue.

[0119] The present invention relates to anti-IL2Rγ antigen-binding proteins, such as antibodies or antigen-binding fragments thereof (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H4H12890P; H4H12899P; H4H12900P; H4H12908P; H4H12913P2; H4H12922P2) The present invention provides a container (for example, a plastic or glass vial having a cap or chromatography column, a hollow needle or syringe cylinder) containing the pharmaceutical formulation, which includes any of the following: H4H12924P2, H4H12926P2, H4H12927P2, H4H12934P2, H4H13538P, H4H13541P, H4H13544P2, or H4H13545P2, or a pharmaceutically acceptable carrier.

[0120] The present invention relates to the anti-IL2Rγ antigen-binding protein of the present invention, for example, an antibody or its antigen-binding fragment (e.g., H4H12857P; H4H12858P; H4H12859P; H4H12863P; H4H12874P; H4H12871P; H4H12884P; H4H12886P; H4H12889P; H This includes combinations of 4H12890P;H4H12899P;H4H12900P;H4H12908P;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2). The anti-IL2Rγ antigen-binding protein and further therapeutic agents may be present in a single composition or in separate compositions. For example, in embodiments of the present invention, the further therapeutic agent is an immunosuppressant. In embodiments of the present invention, further therapeutic agents include anti-TNFα antibodies or binding proteins (e.g., infliximab, adalimumab, etanercept, or golimumab), tacrolimus, cyclosporine, corticoids, prednisolone, methylprednisolone, antithymocyte globulin, alemtuzumab, daclizumab, in vitro photopheresis, mycophenolate mofetil, sirolimus, pentostatin, mesenchymal stem cells, inorimomab, denileukin, multispecific (e.g., bispecific) antibodies or antigen-binding fragments thereof that bind to BCMA (B cell maturation antigen) and CD3, and / or basiliximab.

[0121] Along with further therapeutic agents, anti-IL2Rγ antigen-binding proteins, such as H4H12857P;H4H12858P;H4H12859P;H4H12863P;H4H12874P;H4H12871P;H4H12884P;H4H12886P;H4H12889P;H4H12890P;H4H12899P;H4H12900P;H4H12908P ;H4H12913P2;H4H12922P2;H4H12924P2;H4H12926P2;H4H12927P2;H4H12934P2;H4H13538P;H4H13541P;H4H13544P2;or H4H13545P2 are administered to subjects requiring the aforementioned treatment or prevention of IL2Rγ-mediated diseases. A method for treating or preventing sexually transmitted diseases is part of the present invention.

[0122] The term "together" indicates that the components of the present invention, anti-IL2Rγ antigen-binding proteins, such as antibodies or their antigen-binding fragments, can be formulated in a single composition, for example, for simultaneous delivery, together with another drug such as methotrexate, or they can be formulated separately in two or more compositions (e.g., kits containing each component). Components administered together can be administered to the subject at different times than the other components are administered; for example, each administration can be given asynchronously (e.g., separately or sequentially) at intervals over a given period. Separate components administered together can also be administered sequentially but essentially simultaneously during the same administration session. Furthermore, separate components administered together can be administered to the subject via the same or different routes. [Examples]

[0123] The following examples are provided to give a complete disclosure and explanation of the methods and compositions of the present invention to those skilled in the art, and are not intended to limit the scope of what the inventors consider to be their invention.

[0124] Example 1 Identification and isolation of anti-IL2Rγ antibodies Anti-IL2Rγ antibodies were obtained by immunizing VELOCIMMUNE® mice (i.e., engineered mice containing DNA encoding the variable regions of human immunoglobulin heavy chains and kappa light chains) with an IL2Rγ protein immunogen containing the extracellular sequence (ectodomain) of IL2Rγ.

[0125] In particular, the immunogen, human IL2Rg ecto-mmh, has the following amino acid sequence: [ka] including, • Amino acids (1-240): Human IL2Rg ecto(NP_000197.1, L23-A262), and • Amino acids (241-268): Myc-Myc-hexahistidine tag (underlined) It included.

[0126] Antibody immune responses were monitored using IL2Rγ-specific immunoassays. Fully human anti-IL2Rγ antibodies were isolated and purified.

[0127] [Table 4]

[0128] [Table 5]

[0129] The amino acid sequences of the heavy and light chains of the anti-IL2Rγ antibody immunoglobulin are listed below (CDR is underlined; variable regions are in bold font).

[0130] H4H12857P Heavy chain (SEQ ID NO: 311) [ka]

[0131] Light chain (Sequence ID 313)

change

[0132] H4H12858P Heavy chain (SEQ ID NO: 331)

change

[0133] Light chain (Sequence ID 333)

change

[0134] H4H12859P Heavy chain (SEQ ID NO: 18)

change

[0135] Light chain (Sequence ID 20)

change

[0136] H4H12863P Heavy chain (SEQ ID NO: 38)

change

[0137] Light chain (Sequence number 40)

change

[0138] H4H12874P Heavy chain (SEQ ID NO: 58)

change

[0139] Light chain (Sequence number 60)

change

[0140] H4H12871P Heavy chain (SEQ ID NO: 376)

change

[0141] Light chain (Sequence number 378)

change

[0142] H4H12884P Heavy chain (SEQ ID NO: 77)

change

[0143] Light chain (Sequence ID 79)

change

[0144] H4H12886P Heavy chain (SEQ ID NO: 97)

change

[0145] Light chain (Sequence ID 99)

change

[0146] H4H12889P Heavy chain (SEQ ID NO: 357)

change

[0147] Light chain (Sequence ID 359)

change

[0148] H4H12890P Heavy chain (SEQ ID NO: 115)

change

[0149] Light chain (Sequence number 117)

change

[0150] H4H12899P Heavy chain (SEQ ID NO: 134)

change

[0151] Light chain (Sequence number 136)

change

[0152] H4H12900P Heavy chain (SEQ ID NO: 152)

change

[0153] Light chain (Sequence ID 154)

change

[0154] H4H12908P Heavy chain (SEQ ID NO: 170)

change

[0155] Light chain (Sequence number 172)

change

[0156] H4H12913P2 Heavy chain (SEQ ID NO: 186)

change

[0157] Light chain (Sequence number 188)

change

[0158] H4H12922P2 Heavy chain (SEQ ID NO: 343)

change

[0159] Light chain (Sequence number 188)

change

[0160] H4H12924P2 Heavy chain (SEQ ID NO: 198)

change

[0161] Light chain (Sequence number 188)

change

[0162] H4H12926P2 Heavy chain (SEQ ID NO: 208)

change

[0163] Light chain (Sequence number 188)

change

[0164] H4H12927P2 Heavy chain (SEQ ID NO: 216)

change

[0165] Light chain (Sequence number 188)

change

[0166] H4H12934P2 Heavy chain (SEQ ID NO: 234)

change

[0167] Light chain (Sequence ID 236)

change

[0168] H4H13538P Heavy chain (SEQ ID NO: 254)

change

[0169] Light chain (Sequence number 256) [ka]

[0170] H4H13541P Heavy chain (SEQ ID NO: 272) [ka]

[0171] Light chain (Sequence number 274) [ka]

[0172] H4H13544P2 Heavy chain (SEQ ID NO: 284) [ka]

[0173] Light chain (Sequence number 188) [ka]

[0174] H4H13545P2 Heavy chain (SEQ ID NO: 294) [ka]

[0175] Light chain (Sequence number 188) [ka] *The antibodies referred to in these examples have immunoglobulin chains having the amino acid sequence specifically described in Example 1.

[0176] Example 2 Surface plasmon resonance coupling assay Using the real-time surface plasmon resonance-based Biacore 4000 biosensor platform, the dissociation rate constant (k) for the binding of IL-2Rγ reagent to purified anti-IL2Rγ monoclonal antibody was determined. d The binding was determined. All binding studies were performed at 25°C and 37°C using two running buffers: (i) 1.9 mM NaH2PO4, 8.1 mM Na2HPO4, 2.7 mM KCl, 137 mM NaCl, 0.03% NaN3, 0.05% v / v Surfactant Tween-20, pH 7.4 (PBS-T-pH 7.4), and (ii) 8.8 mM NaH2PO4, 1.2 mM Na2HPO4, 2.7 mM KCl, 137 mM NaCl, 0.03% NaN3, 0.05% v / v Surfactant Tween-20, pH 6.0 (PBS-T-pH 6.0). Anti-IL2Rγ monoclonal antibodies expressed with human IgG4 Fc were captured using a CM5 Biacore sensor surface derivatized by amine coupling with a monoclonal mouse anti-human Fc antibody (GE, catalog #BR-1008-39). All IL2Rγ reagents were C-terminal myc-myc-hex. The extracellular domains were expressed with a histidine tag (hereinafter referred to with the -MMH suffix). Human IL2Rγ extracellular domains expressed with a C-terminal myc-myc-hexahistidine tag (hIL-2Rg-MMH; SEQ ID NO: 379) or cynomolgus monkey IL2Rγ extracellular domains expressed with a C-terminal myc-myc-hexahistidine tag (mfIL-2Rg-MMH; SEQ ID NO: 380) were prepared in PBS-T-pH 7.4 running buffer (100 nM to 11.11 nM; 3-fold serial dilutions) and injected at a flow rate of 30 μL / min for 4 minutes. Dissociation of bound IL-2Rg-MMH was performed in PBS-T-pH 7.4 or PBS-T-pH 6.0 running buffer for 6 minutes.

[0177] By fitting a real-time coupled sensor gram to a 1:1 coupled model using Scrubber 2.0c curve fitting software, the dissociation rate constant (k) in two running buffers is determined. d The following was determined. The dissociation rates of anti-hemoduvelin mAbs for binding to hIL-2RG-MMH and mfIL-2RG-MMH at 25°C and 37°C in PBS-T-pH 7.4 and PBS-T-pH 6.0 are shown in Tables 2-1 to 2-8.

[0178] [Table 6]

[0179] [Table 7]

[0180] [Table 8]

[0181] [Table 9]

[0182] [Table 10]

[0183] [Table 11]

[0184] [Table 12]

[0185] [Table 13]

[0186] Example 3 :Binding kinetics Using the Biacore 4000 instrument equipped with a real-time surface plasmon resonance biosensor, the equilibrium dissociation constant (K) for the binding of IL-2Rγ to purified anti-IL2Rγ monoclonal antibody was determined. D The values ​​were determined. All binding studies were performed at 25°C and 37°C in running buffer of 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% v / v Surfactant Tween-20, pH 7.4 (HBS-ET). The Biacore sensor surface was initially derivatized by amine coupling with monoclonal mouse anti-human Fc antibody (GE, #BR-1008-39) to capture the anti-IL2Rγ monoclonal antibody.

[0187] Binding studies were performed with the following IL-2Rγ reagents:

[0188] • Amino acid sequence: [ka] including, Amino acids (1-240): Human IL2Rg ecto(NP_000197.1, L23-A262) Amino acids (241-268): Myc-Myc-hexahistidine tag (underlined) The human IL2Rγ extracellular domain (hIL-2Rg-MMH; SEQ ID NO: 379) expressed with a C-terminal myc-myc-hexahistidine tag, including the following:

[0189] • Amino acid sequence: [ka] including, Amino acids (1-240): Cynomolgus monkey IL2Rg ecto(L23-A262 of XP_005593949.1) Amino acids (241-268): Myc-Myc-hexahistidine tag (underlined) The extracellular domain of cynomolgus monkey IL2Rγ expressed with a C-terminal myc-myc-hexahistidine tag (mfIL-2Rg-MMH; SEQ ID NO: 380)

[0190] • Amino acid sequence: [ka] including, Amino acids (1-240): Human IL2Rg ecto(NP_000197.1, L23-A262) Amino acids (241-473): Mouse IgG2a Fc tag (underlined) Including human IL2Rγ extracellular domain expressed with a C-terminal mouse IgG2a Fc tag. In (hIL-2Rg-mFc; Sequence ID 381)

[0191] • Amino acid sequence: [ka] including, Amino acids (1-131): Human IL2Rg domain 1 (L23-I153 of NP_000197.1) Amino acids (132-159): Myc-Myc-hexahistidine tag (underlined) The D1 domain of the human IL-2Rγ extracellular domain expressed with a C-terminal myc-myc-hexahistidine tag (hIL-2Rg_D1-MMH; SEQ ID NO: 382)

[0192] • Amino acid sequence: [ka] including, Amino acids (1-88): Human IL2Rg domain 2 (NP_000197.1, P154-S241) Amino acids (89-116): Myc-Myc-hexahistidine tag (underlined) The D2 domain of the human IL2Rγ extracellular domain expressed with a C-terminal myc-myc-hexahistidine tag (hIL-2Rg_D2-MMH; SEQ ID NO: 383)

[0193] • Amino acid sequence: [ka] including, Amino acids (1-241): Mouse IL2Rg ecto(NP_038591.1 W23-A263) Amino acids (242-269): Myc-Myc-hexahistidine tag (underlined) The mouse IL2Rγ extracellular domain (mIL-2Rg-MMH; SEQ ID NO: 384) expressed with a C-terminal myc-myc-hexahistidine tag, including the following:

[0194] • Amino acid sequence: [ka] including, Amino acids (1-240): Rat IL2Rg ecto(NP_543165.1 W23-A262) Amino acids (241-268): Myc-Myc-hexahistidine tag (underlined) The rat IL2Rγ extracellular domain (rIL-2Rg-MMH; SEQ ID NO: 385) expressed with a C-terminal myc-myc-hexahistidine tag, including the following:

[0195] Different concentrations of IL2Rγ reagent were prepared in HBS-ET running buffer (100 nM to 6.25 nM for hIL-2Rg-mFc; 4-fold serial dilution or 50 nM to 3.125 nM; 4-fold serial dilution) and injected onto the surface of anti-human Fc-capturing anti-IL2Rγ monoclonal antibody at a flow rate of 30 μL / min for 4 minutes. Dissociation of the monoclonal antibody-bound IL2Rγ reagent was monitored in HBS-ET running buffer for 8 to 10 minutes. Kinetic association (k) was determined by fitting the real-time sensorgram to a 1:1 binding model using Scrubber 2.0c curve fitting software. a ) and dissociation (k d The rate constant was determined. The bond dissociation equilibrium constant (K D ) and the dissociation half-life (t1 / 2)

number

[0196] Tables 3-1 to 3-14 show the kinetic parameters for the binding of various IL-2Rγ reagents to different IL-2Rγ monoclonal antibodies at 25°C and 37°C.

[0197] [Table 14]

[0198] [Table 15]

[0199] [Table 16]

[0200] [Table 17]

[0201] [Table 18]

[0202] [Table 19]

[0203] [Table 20]

[0204] [Table 21]

[0205] [Table 22]

[0206] [Table 23]

[0207] [Table 24]

[0208] [Table 25]

[0209] [Table 26]

[0210] [Table 27]

[0211] Example 4 Octet cross-competition between different anti-IL-2Rγ monoclonal antibodies The competition for binding between a series of anti-IL2Rγ monoclonal antibodies was determined using a real-time label-free biolayer interferometry assay on the Octet HTX biosensor platform (Pall ForteBio Corp.). The entire experiment was performed at 25°C in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% v / v Surfactant Tween-20, 1 mg / mL BSA, pH 7.4 (HBS-EBT) buffer using a plate shaker speed of 1000 rpm. The competition for binding between the two antibodies was determined on the human IL2Rγ extracellular domain (hIL-2Rg-MMH; SEQ ID NO: 379) expressed with a C-terminal myc-myc-hexahistidine tag. To evaluate whether the antibodies compete with each other for binding to the epitope, an anti-pentaHis antibody-coated Octet biosensor tip (Fortebio Inc, #18-5122) was used to capture approximately 0.27 nM of hIL-2Rg-MMH by immersing the tip in a well containing 10 μg / mL of hIL-2Rg-MMH for 3 minutes. The antigen-captured biosensor tip was then saturated with a first anti-IL2Rγ monoclonal antibody (hereinafter referred to as mAb-1) by immersing it in a well containing 50 μg / mL of mAb-1 for 300 seconds. The biosensor tip was then immersed in a well containing 50 μg / mL of a second anti-IL2Rγ monoclonal antibody (hereinafter referred to as mAb-2) for 240 seconds. The biosensor tip was washed in HBS-ETB buffer between each step of the experiment. The binding response was monitored in real time throughout the entire experiment, and the binding response was recorded at the end of each step. The response of mAb-2 binding to hIL-2Rg-MMH pre-complexed with mAb-1 was compared, and the competitive / non-competitive behavior of different anti-IL2Rγ monoclonal antibodies was determined as shown in Table 4-1.

[0212] [Table 28] [Table 29] [Table 30]

[0213] Example 5 Flow cytometry analysis of STAT phosphorylation in human CD4+ T cells (human PBMCs) To evaluate the in vitro characteristics of the IL2Rγ antibody of the present invention, CD4 induced by IL-2, IL-4, IL-7, IL-15, and IL-21 were used. + Their ability to block T cell activation is measured by flow cytometry (BD(trademark) Phosflow). Measured by (Say). BD(trademark) Phosflow is a phosphoprotein in cells. This technology allows for the simultaneous analysis of cellular signaling in separate subpopulations of cells by analyzing (such as STAT proteins) and cell surface markers. Using this technique, human CD4 stimuli with cytokines from the gamma c family can be analyzed. + We analyzed STAT phosphorylation in T cells.

[0214] Human peripheral blood mononuclear cells (PBMCs) were isolated from fresh whole blood (Bioreclammation IVT) by density gradient centrifugation. K2 EDTA whole blood was then subjected to X-VIVO™. Diluted 1:1 in medium (Lonza), added to a SepMate tube (StemCell) containing FicollPaquePLUS (Healthcare), and centrifuged to separate the PBMCs. The upper layer containing the PBMCs was transferred to a new tube and washed twice with DPBS (Life Technologies). The PBMCs were then separated into approximately 5.0 × 10⁶ units. 6 Resuspend in X-VIVO® 15 medium at a concentration of cells / mL and prepare 96 wells. The cells were plated (50 μL per well; approximately 250,000 cells per well), incubated at 37°C for 2 hours, and then cytokines and antibodies were added.

[0215] Starting with a final antibody concentration of 400 nM, serial dilutions of the antibody (1:5) were prepared in pre-warmed X-VIVO® 15 medium and added to cells (50 uL). IL-7(R& Final concentrations of 1 pM for D Systems, 50 pM for IL-4 (R&D Systems) and IL-21 (eBioscience), 0.5 nM for IL-15 (R&D Systems), and 10 nM for IL-2 (R&D Systems) were prepared in pre-warmed X-VIVO® 15 medium at a final volume of 200 μL / well and added to cells (100 μL).

[0216] For cytokine dose response, serial dilutions (1:5) of each cytokine, starting at 5 nM for IL-4, IL-7, and IL-21, or 50 nM for IL-2 and IL-15, are also preheated in X-VIVO® 1. The preparation was carried out in 5 culture media. First, 50 μL of X-VIVO® 15 medium was added to the cells, with a total volume of 200 μL per well, followed by serial dilutions of 100 μL of cytokines. I added the liquid.

[0217] After adding cytokines and antibodies to the cells, they were incubated at 37°C for 15 minutes to activate the PBMCs (STAT phosphorylation). The stimulation was then terminated by adding 200 μL of warm Cytofix (BD) to each well, and the cells were incubated at 37°C for 10 minutes (fixation step). The cells were then washed twice with staining buffer (BD) and kept at 4°C overnight. The following day, the cells were centrifuged and permeabilized by slowly adding 100 μL of cold Perm Buffer III (BD) to the pellet. The cells were incubated at 4°C for 30 minutes and then washed twice with staining buffer. CD4 was used to measure STAT phosphorylation. +To enable the analysis of T cell populations, human FcR binding inhibitors (eBioscience; 1 / 10), anti-CD33-PE (BD; 1 / 200), anti-CD4-PacificBlue (BD; 1 / 200), anti-CD3-PECy7 (BD; 1 / 200), and related anti-phospho-STAT-AlexaFluor647 (BD) were prepared in staining buffer: - Anti-phospho STAT3 (1 / 10): For cells stimulated with IL-21, - Anti-phospho STAT5(1 / 20): For cells stimulated with IL-2, IL-7 and IL-15, - AntiphosphoSTAT6(1 / 10): For cells stimulated with IL-4 The cells were stained using a mixture of the following:

[0218] The sample was kept in the dark for 1 hour. The cells were then centrifuged and washed twice with staining buffer. Sample data was acquired on an LSR Fortessa X-20 cell analyzer using the HTS attachment (BD). Data analysis was performed using FlowJo X software (Tree Star, OR). CD4 + T cells are intact cells, singlets, and CD33 cells. - CD3 + CD4 + We defined STAT phosphorylation as such and analyzed it within this cell population (MFI = mean fluorescence intensity).

[0219] Both H4H12889P and H4H12922P2 similarly and efficiently blocked STAT phosphorylation induced by all cytokines tested in this assay (IL-2, IL-4, IL-7, IL-15, and IL-21). However, in addition to H4H12874P, H4H12886P, and H4H12857P, the comparative antibody COMP1499 (anti-IL2Rγ antibody CP.B8, see US2002 / 0028202) partially blocked or did not block cytokine-induced STAT phosphorylation.

[0220] [Table 31]

[0221] See also Figure 1(A-E), which shows the levels of STAT phosphorylation at each concentration of the tested antibodies.

[0222] Example 6 Flow cytometry analysis of STAT3 phosphorylation in differentiated human mast cells in vitro To evaluate the in vitro characteristics of the anti-IL2Rγ antibody of the present invention, their ability to block IL-9-induced human mast cell activation was measured by flow cytometry (BD® Phosflow assay). This technique was used to investigate STAT3 phosphorylation in in vitro differentiated human mast cells.

[0223] In short, bone marrow CD133 cultured for 6 weeks in StemSpan serum-free medium supplemented with human SCF, IL-6, and IL-3. + Human mast cells were generated in vitro from progenitor cells.

[0224] Human mast cells are approximately 4.0 × 10 6 In X-VIVO® 15 medium at a concentration of cells / mL The cells were resuspended, plated into 96-well plates (50 μL of cells per well; approximately 200,000 cells per well), incubated at 37°C for 2 hours, and then cytokines and antibodies were added.

[0225] Starting with a final antibody concentration of 400 nM, serial dilutions of the antibody (1:5) were prepared in pre-warmed X-VIVO® 15 medium and added to cells (50 uL). The final concentration was 2 nM. The fixed IL-9 (R&D) concentration was prepared in pre-warmed X-VIVO (trademark) 15 medium at a final volume of 200 μL / well and added to cells (100 μL).

[0226] For cytokine dose response, serial dilutions of IL-9 (1:5) are also prepared in pre-warmed X-VIVO® 15 medium, starting at a final cytokine concentration of 100 nM. It was prepared. First, with a total volume of 200uL per well, 50uL of X-VIVO( (Trademark) 15 medium was added to the cells, followed by 100 uL of serially diluted cytokine solution.

[0227] After adding cytokines and antibodies to the cells, they were incubated at 37°C for 15 minutes to activate the mast cells (measured by STAT3 phosphorylation). Next, the stimulation was terminated by adding 200 μL of warm Cytofix (BD) to each well, and the cells were incubated at 37°C for 10 minutes (fixation step). The cells were then washed twice with staining buffer (BD) and kept at 4°C overnight. The following day, the cells were centrifuged and permeabilized by slowly adding 100 μL of cold Perm Buffer III (BD) to the pellet. The cells were incubated at 4°C for 30 minutes and then washed twice with staining buffer. The mast cells were then stained with a mixture of human FcR binding inhibitor (eBioscience; 1 / 10), anti-c-Kit-PE (BD; 1 / 100), and anti-phospho-STAT3-AlexaFluor647 (BD; 1 / 10) prepared in staining buffer.

[0228] The sample was kept in the dark for 1 hour. The cells were then centrifuged and washed twice with staining buffer. Sample data was acquired on an LSR Fortessa X-20 cell analyzer using the HTS attachment (BD). Data analysis was performed using FlowJo X software (Tree Star, OR). Mast cells were analyzed as intact cells, singlets, and c-Kit cells. + We defined STAT3 phosphorylation as such and analyzed it within this cell population (MFI = mean fluorescence intensity).

[0229] Both H4H12889P and H4H12922P2 similarly and efficiently blocked IL-9-induced STAT3 phosphorylation.

[0230] [Table 32]

[0231] See also Figure 2, which shows the level of IL-9-induced STAT phosphorylation at each concentration of the tested antibodies.

[0232] Example 7 Testing of monoclonal antibodies in in vivo models; xenograft-versus-host disease model to evaluate the blockade of IL-2R gamma antibody activity as a therapeutic measure. In relation to in vivo models, xenograft-versus-host disease (GvHD) studies were conducted to determine the effects of our anti-IL2Rγ antibodies, H4H12889P and H4H12922P2, along with the comparative IL-2Rγ antibody COMP1499. Briefly, to induce GvHD in mice, human peripheral blood mononuclear cells (huPBMCs) were treated with NOD-scid IL2Rγ null The drug was injected into (NSG) mice (Jackson Lab). Upon transplantation, human immune cells recognize the mouse host as a heterologous organism and initiate an active immune response against its tissues.

[0233] In this experiment, DPBS was applied to the posterior orbit of NSG mice (Jackson Lab). Ten million resuspended human PBMCs (ReachBio) were injected (10 million cells / 100uL; 5 groups of 10 mice each). Briefly, on the day of injection, human PBMCs were thawed in IMDM medium (Irvine Scientific) supplemented with 10% FBS (Seradigm) and incubated in this supplemented medium at 37°C for 2 hours. The cells were then washed in DPBS (Life Technologies) and resuspended at 10 million cells / 100uL for injection. 100uL of PBS was injected into the posterior orbit of the control group (10 mice). Four groups of huPBMC-transplanted NSG mice were subcutaneously injected with either 25 mg / kg of H4H12889P, H4H12922P2, COMP1499, or an isotype control antibody (REGN1945; human anti-domestic cat (Felis domesticus) Fel d1 antibody (IgG4(S108P) / kappa)) twice weekly for 6 weeks, starting 3 weeks after huPBMC injection. The experiment was terminated on day 161 after huPBMC transplantation by sacrificing the remaining mice. The experimental drug administration and treatment protocols for each mouse group are shown in Table 7-1.

[0234] [Table 33]

[0235] Throughout the experiment, mice were monitored twice a week for weight loss and death (to assess the effect of therapeutic antibodies on survival). In addition to human cell transplantation in the blood, serum mouse and human cytokine levels were assessed at different time points, as shown in Table 7-2.

[0236] [Table 34]

[0237] Throughout the experiment, mice were monitored twice a week for weight loss (Figure 3(A-F); %) of initial body weight on the day of huPBMC transplantation and death (Figure 4; to evaluate the effect of therapeutic antibodies on survival). Animals showing a weight loss of 20% of their initial body weight were euthanized.

[0238] Human cell transplantation was evaluated by collecting blood samples from mice at different time points after huPBMC injection into Microtainer tubes (BD, Cat#3659740) and examining the absolute number of human cells in the blood by flow cytometry. Briefly, 50 μL of each blood sample was incubated in ACK lysis buffer (Gibco) at room temperature for 5 minutes to lyse erythrocytes. The cells were then washed in DPBS, stained with LIVE / DEAD fixable dead stain (Invitrogen), washed in MACS buffer (Miltenyi Biotec), and then stained with human CD45. + cell, T cell, CD4 + T cells and CD8 + T cells were labeled using a mix of antibodies used for identification (anti-human CD45, anti-human CD3, anti-human CD4, and anti-human CD8 [BD] diluted 1 / 50 in brilliant staining buffer [BD], along with human and mouse Fc inhibitor antibodies [eBioscience and BD, respectively]). Finally, the samples were washed in MACS buffer, fixed in BD CytoFix (BD), and then resuspended in MACS buffer containing CountBright beads (Life Technologies) to calculate the absolute number of cells in each sample. Sample data was acquired on an LSR Fortessa X-20 cell analyzer using an HTS attachment (BD). Data analysis was performed using FlowJo X software (Tree Star, OR). Human CD45 + T cells, living cells, singlet, CD45 + Defined as such, and within this group CD4 + T cells and CD8 + T cells are each CD3 + CD4 + and CD3 + CD8+ It was further defined as follows.

[0239] [Table 35]

[0240] As an example, Figure 5 (A-D) shows the absolute number of human cells in the blood 35 days after huPBMC injection. Human CD45+ cells, T cells, CD4+ T cells, and CD8+ cells over time. The number of T cells is shown in Figure 6 (A-D).

[0241] Serum was collected from mice on different days after huPBMC injection, and serum levels of mouse and human cytokines were evaluated. Briefly, whole blood was collected in Microtainer tubes (BD, Cat#365967) and allowed to coagulate by standing at room temperature for at least 30 minutes. The coagulated blood and cells were pelleted by centrifugation at 15,000 × g for 10 minutes at 4°C. The resulting supernatant, designated as serum, was transferred to a clean plate, and cytokine concentrations in the serum were measured using two Proinflammatory (mouse and human) multiplex immunoassay kits (Meso Scale Discovery) according to the manufacturer's instructions. The plates were washed with PBS containing 0.05% (w / v) Tween-20 (Life Technologies). Electrochemiluminescence was immediately read using an MSD Spector instrument. Data analysis was performed using FlowJo X software (Tree Star, OR).

[0242] [Table 36]

[0243] [Table 37]

[0244] As an example, Figure 7(A-I) shows serum human and mouse cytokine levels 42 days after huPBMC injection. Figure 8(A-D) shows serum levels of human IFN-γ, human TNFα, mouse TNFα, and mouse IL-6 over time.

[0245] This in vivo study demonstrated the efficacy of the anti-IL2Rγ antibodies, H4H12889P and H4H12922P2, when therapeutically administered in a model of graft-versus-host disease. While COMP1499 did not, both H4H12889P and H4H12922P2 efficiently blocked the development of GvHD in mice. Mice therapeutically treated with either of these two antibodies were protected from weight loss and death, accompanied by a dramatic reduction in both mouse and human serum cytokine levels in the blood, as well as human T cell counts. See Tables 7-3, 7-4, and 7-5.

[0246] Example 8 Bioassay using NK92 / hIL7R / STAT3-Luc and Ramos.2G6.4C10 / STAT3-Luc cells The IL2Rγ family cytokines, IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21, transmit signals via the JAK-STAT (Janus kinase-signal transducer and transcriptional activator) pathway (Rochman et al., New insights into the regulation of T cells by gamma(c)family cytokines. Nat Rev). Immunol. 2009 Jul;9(7):480-490). To evaluate the inhibition of cytokine signaling by anti-IL2Rγ antibodies, a bioassay was developed using NK-92 cells (human natural killer cell line, ATCC) that stably express a luciferase reporter (STAT3-Luc; SABiosciences, #CLS-6028L). NK-92 cells endogenously expressed ligand-selective receptors that mediate the signaling of IL2Rγ as well as IL-2, IL-9, IL-15, and IL-21. To also evaluate the regulation of IL-7 signaling, NK-92 cells were transduced using a lentivirus containing human IL-7R, and stably expressing cells were selected and maintained in G418. The resulting cell line will hereafter be referred to as NK-92 / hIL7R / STAT3-Luc. To investigate the regulation of IL-4-mediated signaling, Ramos.2G6.4C10 (human B lymphocyte cell line, ATCC) cells, which endogenously express IL2Rγ and IL-4R receptors, were transduced with a STAT3-luc reporter. The resulting cell line was named Ramos.2G6.4C10 / STAT3-Luc.

[0247] The anti-IL2γ antibody of the present invention was tested for inhibition of human IL-2 (hIL-2), human IL-7 (hIL-7), human IL-9 (hIL-9), human IL-15 (hIL-15), or human IL-21 (hIL-21) signaling by plating 20,000 NK-92 / hIL7R / STAT3-Luc cells / well in growth medium (prepared according to the instructions for use by ATCC, but without IL-2) in 96-well plates and incubating overnight at 37°C in 5% CO2. The following day, the anti-IL2Rγ antibody or isotype control was serially diluted in assay buffer from 500 to 0.008 nM (in addition, samples containing the buffer alone without the test molecule) and added to the cells, and incubated for 30 minutes. After incubation, ligands were added to cells at the following final concentrations: 30 pM hIL-2, 50 pM hIL-7, 20 pM hIL-9, 60 pM or 100 pM hIL-15, or 5 pM or 3 pM hIL-21. Dose-dependent activation was determined using serial dilutions of 10 nM to 0.2 pM ligands added to cells (in addition, samples containing buffer alone without ligands). After incubation at 37°C in 5% CO2 for 5 hours, OneGlo® reagent (Promega, #E6031) and a Victor® X multi-label plate reader (Perkin Elme) were used. Luciferase activity was measured using (r).

[0248] To test the anti-IL2γ antibody of the present invention in inhibiting human IL-4 (hIL-4) signaling, Ramos.2G6.4C10 / STAT3-Luc cells were plated in growth medium (prepared according to instructions for use by ATCC) at a density of 100,000 cells / well in a 96-well plate. Anti-IL2Rγ antibody or isotype control was serially diluted in assay buffer from 500 to 0.008 nM (plus samples containing the buffer alone without the test molecule) and added to the cells, and incubated for 20 minutes. After incubation, hIL-4 was added to the cells at a final concentration of 250 pM or 200 pM. Dose-dependent activation was determined using serial dilutions of 10 nM to 0.2 pM hIL-4 added to the cells (plus samples containing the buffer alone without the ligand). After overnight incubation at 37°C in 5% CO2, the samples were processed using OneGlo® reagent (Promega, #E6031) and a Victor® X multi-label plate reader (Perkin E). Luciferase activity was measured using LMER.

[0249] Nonlinear regression (4 parameters) using Prism 5 software (GraphPad) - Using logistics) to analyze results and EC 50 and IC 50 The value was obtained. The following equation:

number

[0250] In this equation, "RLU ベースライン "RLU" is the luminescence value from cells treated with a fixed amount of ligand without the use of antibodies. 阻害 "RLU" is the minimum luminescence value from cells treated with a dose response of a specific antibody at a specific ligand concentration. バックグラウンド " represents the luminescence value from cells treated without any ligands or antibodies.

[0251] [Table 38]

[0252] [Table 39]

[0253] The 23 anti-IL2γ antibodies of the present invention were tested using bioassays for their ability to inhibit cytokine-mediated signaling of the IL2Rγ family. As shown in Table 8-1, 19 of the 23 anti-IL2γ antibodies inhibited IL2Rγ activation to varying degrees, and as shown in Table 8-2, 4 of the 23 anti-IL2γ antibodies did not show ligand-mediated inhibition of IL2Rγ activation.

[0254] Example 9 Cell-to-cell binding analysis by flow cytometry using NK-92, Jurkat, NIH / 3T3, MC / 9, and HEK293 cells. To evaluate the binding of anti-IL2Rγ antibodies to human and mouse IL-2Rγ expressed on cells, flow cytometry analysis was performed using cell lines that endogenously express IL-2Rγ: NK-92 (human natural killer cell line), Jurkat (human T lymphocyte cell line), and MC / 9 (mouse mast cell line). NIH / 3T3 (mouse fibroblast) and HEK293 (human fetal kidney) cell lines were included as negative controls.

[0255] For flow cytometry analysis, cells were pre-incubated with 100 μg / ml mouse IgG at room temperature (RT) for 15 minutes to block antibody binding to the Fc receptor. Jurkat, NIH / 3T3, and HEK293 cells were incubated in PBS (calcium and magnesium-free) containing 1% FBS, or NK-92 and MC / 9 cells were incubated in growth medium (prepared according to instructions for use by ATCC) at a rate of 0.5–1 × 10⁶. 6Each cell type in the cell / well was treated with the anti-IL2Rγ antibody and isotype control antibody of the present invention at 10 μg / ml for 30–45 minutes at RT. The cells were washed and incubated with allophycocyanin (APC)-conjugated anti-human antibody (Jackson ImmunoResearch, #109-136-170) on ice for 30 minutes. The cells were washed, fixed using BD CytoFix® (BD biosciences, #554655), and IQue® (Intellicyt®) Flow C Analysis was performed on a cytometer or Accuri Flow cytometer (BD). Unstained and secondary antibody-only controls were also included for all cell lines. The results were analyzed using t(registered trademark) (IntelliCyt(registered trademark)) software to determine the geometric mean (MFI) of fluorescence for viable cells. The binding ratio was calculated by normalizing the MFI of the test sample using the MFI of the unstained sample.

[0256] As shown in Table 9-1, 19 of the 23 anti-IL2Rγ antibodies of the present invention tested at 10 μg / ml demonstrated binding to Jurkat and NK-92 cells at binding ratios of 1-19 and 1-94, respectively. The anti-IL2Rγ antibodies demonstrated binding to NIH / 3T3 and MC / 9 cells at binding ratios of 1-13 and 1. Human isotype control antibody, REGN1945, and secondary single control conditions exhibited binding ratios of 1-13 for all cell lines tested.

[0257] As shown in Table 9-2, four of the 23 anti-IL2Rγ antibodies of the present invention tested at 10 μg / ml demonstrated binding to NK-92 cells at binding ratios of 1–37 and to HEK293 cells at binding ratios of 1–3. Human isotype control antibody, REGN1945, and secondary single control conditions exhibited binding ratios of 1–2 to NK-92 and HEK293 cells, respectively.

[0258] [Table 40]

[0259] [Table 41]

[0260] Example 10 In vivo immunosuppression experiment to evaluate the effect of the anti-IL2Rγ antibody H4H12889P on immune cell populations in the blood. Experimental procedure. Velocigene® (VG) background mice (C57BL / 6NTac (75%) / 129S6SvEvTac (25%)) from Regeneron Velocigene® breeding colonies genetically modified to replace the endogenous IL2RG ectodomain with the corresponding human sequence were administered subcutaneously with or without isotype control (REGN1945) or H4H12889P at doses of 10 mg / kg or 25 mg / kg twice weekly for 3 weeks (total of 6 doses).

[0261] [Table 42]

[0262] Analysis of immune cell populations in blood over time using flow cytometry. Total immune cells, B cells, T cells, NK cells, and neutrophil counts in peripheral blood were analyzed via flow cytometry at various time points (weekly) to evaluate the effect of H4H12889P on the absolute numbers of these cell types. Briefly, at each time point, blood samples from mice were collected in Microtainer tubes containing K2EDTA [BD #365974], and 30–75 μL of each blood sample was incubated in erythrocyte lysis buffer [Sigma #R7757] at room temperature for 5 minutes to lyse erythrocytes. A second round of lysis was performed if necessary. The cells were then washed in DPBS [Gibco #14190-144] and LIVE / DEAD® Fixable N240P was diluted 1:500 in DPBS. Stain with ear-IR Dead Cell Stain [Invitrogen #L34962] for 20 minutes, wash again in DPBS, then MACS buffer [autoMACS Running Buffer; Miltenyi Biotec, #130] The cells were blocked with purified anti-mouse CD16 / CD32 (Fc Shield) [Tonbo Biosciences, #70-0161-M001] diluted 1:50 in [-091-221]. Subsequently, the cells were stained for cell surface markers and BD Adding a mixture of fluorescently labeled antibodies (listed in Table 2) diluted in horizon brilliant staining buffer [BD #566349] results in CD45 + Cells, T cells, B cells, NK cells, and neutrophils were identified. Finally, samples were washed in MACS buffer, fixed in BD CytoFix [BD #554655] diluted 1:4 in DPBS, and then acquired after washing and resuspension in MACS buffer. Sample data was acquired on a FACSymphony A5 analyzer using an HTS attachment [BD]. A constant volume of each sample was flowed. Data analysis was performed using FlowJo v10 software [Tree Star, OR]. CD45 + Immune cells are called singlets, live cells, and CD45 cells. + Defined as; within this population, T cells are CD3 + As such, B cells CD3 - CD19 + As such, NK cells CD3 - CD19 - NKp46 + For example, neutrophils F4 / 80 - Ly6G + It was further defined as follows: The number of cells per μL of blood for each sample was calculated using the absolute number of each cell type passed through the analyzer, the volume of sample passed through, and the volume of originally stained blood.

[0263] [Table 43]

[0264] Analysis of serum therapeutic antibody levels over time using antigen capture ELISA. Serum levels of IL2Rγ antibody or isotype control antibody were measured weekly using the Human Total IgG Platinum ELISA kit. Serial dilutions of each antibody were prepared in a 0.5% solution of BSA in PBS, and standard curves were generated from H4H12889P and REGN1945 at concentrations of 1.56–100 ng / mL. Absorbance at 450 nm was measured on a SpectraMax M5 plate reader [Molecular Devices]. Data analysis was performed using Prism 8.1.2 [GraphPad].

[0265] Summary of results and conclusions. Treatment with H4H12889P (10 mg / kg and 25 mg / kg) improved total CD45 levels in the blood. + This resulted in a significant reduction in the numbers of immune cells (Figure 9(A)), NK cells (Figure 9(B)), T cells (Figure 9(C)), and B cells (Figure 9(D)), but there was no effect on neutrophil count (Figure 9(E)). After the completion of the 3-week treatment period, serum concentrations of H4H12889P decreased over time. This decrease in H4H12889P concentration was attributed to total CD45 + This was accompanied by a continuous increase in the number of immune cells (Figure 9(A)), NK cells (Figure 9(B)), T cells (Figure 9(C)), and B cells (Figure 9(D)). By the end of the study, all of these populations had recovered to levels similar to those observed before treatment and in mice that were untreated or treated with REGN1945 (isotype control).

[0266] Example 11 : An in vivo skin graft rejection model to evaluate the blockade of the activity of the IL2Rγ antibody H4H12889P. Experimental procedure. BALB / cJ mice obtained from The Jackson Laboratory (Bar Harbor, ME) were used as skin graft donors, and MHC mismatched Velocigene® (VG) background mice (C57BL / 6NTac (75%) / 129S6SvEvTac (25%)) from Regeneron Velocigene® breeding colonies, genetically modified to replace the endogenous IL2RG ectodomain with the corresponding human sequence, were used as recipients. Skin grafts were obtained from the tails of the donor mice. The skin was delaminated using forceps and punched using a 10 mm diameter biopsy punch.

[0267] VG mice (humanized for IL2Rγ) used as graft recipients were administered subcutaneously with either an isotype control (REGN1945) or H4H12889P at a dose of 25 mg / kg twice weekly, starting 3 weeks prior to transplantation and continuing until rejection, or not. Shaved recipients were anesthetized with isoflurane via nose cone and administered an analgesic (buprenorphine - sustained-release) (ZooPharm). Povidone-iodine and alcohol were applied to the shaved dorsal compartment. A graft bed was created midway between the dorsal and ventral sides of the mouse by pinching the skin with forceps and then excising the skin using a sterile 10 mm diameter biopsy skin punch. The graft was then placed on the graft bed, covered with an adhesive bandage, and secured to the skin with two sterile surgical staples. Sterilization techniques were performed throughout the entire procedure. Five days later, the bandages and staples were removed, followed by monitoring.

[0268] [Table 44]

[0269] The experimental design is shown in Figure 10.

[0270] Monitoring of skin graft rejection. Skin graft monitoring included the following criteria: (1) Skin grafts that did not properly angiogenically form were considered technical failures and excluded from analysis. These grafts showed scabbing and shrinkage within a few hours of bandage removal. (2) Scabbing and shrinkage of the graft at later time points were used as indicators of graft rejection. Complete rejection was recorded as the first day when 100% of the graft tissue had necrotized (Figure 12). Onset of rejection was recorded as the first day when signs of rejection (i.e., redness) were present (Figure 11). Significance was determined by the log-rank (Mantel-Cox) test with Bonferroni correction (corrected p-value 0.005, K=9).

[0271] Detection of donor-specific antibodies by flow cytometry. Blood was collected 56 days after transplantation. Donor-specific antibody formation was evaluated by pulling (Figure 13).

[0272] CT26.WT (ATCC(registered trademark) CRL-2638(trademark)) cells, 80% concentration Cells were cultured in tissue culture flasks until fluent. The cells were washed with 1× DPBS, incubated with TrypLE Express reagent (Gibco) at room temperature for 5 minutes, and dissociated by washing the flask with complete RPMI 1640 medium. Next, the cells were centrifuged (500g, 10 minutes) and resuspended at 5 million cells / ml at room temperature for 15 minutes in 1× DPBS with a 1:50 dilution of 4ug / ml Fc block (Tonbo). The suspension was plated in 384-well V-bottom plates at 250,000 cells / well (50uL).

[0273] 50 µl serially diluted serum samples from transplanted mice and non-transplanted wild-type VG mice (C57BL / 6NTac (75%) / 129S6SvEvTac (25%)) as well as from wild-type BALB / cJ mice obtained from The Jackson Laboratory were added to each well and incubated at 37°C for 45 minutes. After two washes with MACS buffer (500 g, 4 mins), 50 µl of LIVE / DEAD® Fixable Blue diluted 1:500 in 1 × DPBS was added to a total volume of 50 µl / well. Cells were resuspended in Dead Cell Stain Kit (Invitrogen) and incubated at room temperature for 15 minutes. After centrifugation at 500g for 4 minutes, the supernatant was discarded, and the cells were resuspended in 25µl of Fc Block (Tonbo) and incubated at 4°C for 15 minutes. Next, 25µl of 2× antibody cocktail (Table 11-2) was added and incubated at 4°C for 25 minutes. After centrifugation (500g, 4 minutes), cells were washed in MACS buffer by adding 100µl of MACS® buffer to each well. 100µl of Cytofix® Fixation Buffer diluted 1:4 in 1× DPBS was added. Cells were fixed by resuspending them in fer(BD) and incubated at 4°C for 15 minutes. After centrifugation and discarding the fixative, the samples were resuspended in MACS buffer. Cells were acquired using a BD Fortessa X-20. Acquired events were analyzed using FlowJo(BD). MFI was derived from doublet-recognized (FSC-H, FSC-A) and live / dead dye-negative cells. The plotted results were median fluorescence intensity values ​​at a 1 / 512 dilution of the sample serum.

[0274] [Table 45]

[0275] Summary of results and conclusions. In the skin graft model (BALB / cJ to VG mice), H4H12889P (anti-IL2Rγ Ab) treatment delayed the onset of skin graft rejection and improved overall skin graft survival. H4H12889P treatment also prevented the generation of donor-specific antibodies in this graft model.

Claims

1. An isolated antibody or antigen-binding fragment that specifically binds to interleukin-2 receptor gamma (IL2Rγ), wherein the antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) containing three heavy chain complementarity-determining regions (CDR-H1, CDR-H2, and CDR-H3) and a light chain variable region (LCVR) containing three light chain complementarity-determining regions (CDR-L1, CDR-L2, and CDR-L3), wherein CDR-H1 comprises the amino acid sequence of SEQ ID NO: 337, CDR-H2 comprises the amino acid sequence of SEQ ID NO: 339, CDR-H3 comprises the amino acid sequence of SEQ ID NO: 341, CDR-L1 comprises the amino acid sequence of SEQ ID NO: 72, CDR-L2 comprises the amino acid sequence of SEQ ID NO: 54, and CDR-L3 comprises the amino acid sequence of SEQ ID NO:

184.

2. An antibody, as described in claim 1, or an antigen-binding fragment thereof.

3. The antibody or its antigen-binding fragment according to claim 2, wherein the antibody is a human antibody.

4. The antibody or its antigen-binding fragment according to claim 1, which is an antigen-binding fragment of an antibody.

5. The antibody or antigen-binding fragment according to any one of claims 1 to 4, wherein HCVR comprises the amino acid sequence of SEQ ID NO: 335, and / or LCVR comprises the amino acid sequence of SEQ ID NO:

182.

6. The antibody or antigen-binding fragment according to any one of claims 1 to 4, wherein HCVR comprises the amino acid sequence of SEQ ID NO: 335, and LCVR comprises the amino acid sequence of SEQ ID NO:

182.

7. An antibody or antigen-binding fragment according to any one of claims 1 to 6, comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 343 and / or a light chain containing the amino acid sequence of SEQ ID NO:

188.

8. A heavy chain containing the amino acid sequence of SEQ ID NO: 343 and the amino acid sequence of SEQ ID NO: 188 An antibody or antigen-binding fragment according to any one of claims 1 to 6, comprising a light chain.

9. The antibody or antigen-binding fragment according to any one of claims 1 to 4, wherein HCVR comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 335, and / or LCVR comprises an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO:

182.

10. An antibody or antigen-binding fragment according to any one of claims 1 to 4, comprising a heavy chain containing an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO: 343 and / or a light chain containing an amino acid sequence having at least 90% sequence identity with respect to the amino acid sequence of SEQ ID NO:

188.

11. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, which is multispecific.

12. The complex comprises an antibody according to any one of claims 1 to 11 or an antigenic fragment thereof, which is bound to an IL2Rγ polypeptide or an antigenic fragment thereof.

13. A method for producing an antibody or antigen-binding fragment according to any one of claims 1 to 11, (a) introducing one or more polynucleotides encoding the HCVR and LCVR of the antibody or its antigen-binding fragment into a host cell; and (b) Culturing host cells under conditions favorable for polynucleotide expression; The method, including the method described above.

14. (c) The method according to claim 13, further comprising isolating an antibody or an antigen-binding fragment from host cells and / or a culture medium in which the host cells were grown.

15. The method according to claim 13 or 14, wherein the host cell is a Chinese hamster ovary cell.

16. An antibody or an antigen-binding fragment thereof, which is a product of the method according to any one of claims 13 to 15.

17. A polynucleotide comprising an antibody or an antigen-binding fragment according to any one of claims 1 to 11.

18. A vector comprising the polynucleotide described in claim 17.

19. A host cell comprising an antibody or antigen-binding fragment thereof, polynucleotide, or vector as described in any one of claims 1 to 11 or 16 to 18.

20. A composition or kit comprising an antibody or antigen-binding fragment according to any one of claims 1 to 11 and 16.

21. The composition or kit according to claim 20, further comprising a further therapeutic agent.

22. A pharmaceutical preparation comprising an antibody or antigen-binding fragment according to any one of claims 1 to 11 and 16, and a pharmaceutically acceptable carrier or excipient.

23. The pharmaceutical formulation according to claim 22, further comprising a further therapeutic agent.

24. The composition or kit according to claim 21 or the formulation according to claim 23, wherein the further therapeutic agent is an anti-inflammatory agent.

25. The composition or kit according to claim 21 or the formulation according to claim 23, wherein the further therapeutic agent is one or more members selected from the group consisting of infliximab, adalimumab, etanercept, golimumab, prednisolone, methylprednisolone, antithymocyte globulin, alemtuzumab, daclizumab, tacrolimus, cyclosporine, mycophenolate mofetil, sirolimus, pentostatin, mesenchymal stem cells, inorimomab, denileukin, and basiliximab.

26. The composition or kit according to claim 21 or the formulation according to claim 23, wherein the further therapeutic agent is one or more members selected from the group consisting of an anti-TNFα antibody or binding protein and corticoids.

27. A container or injection device comprising an antibody or antigen-binding fragment thereof, composition or formulation according to any one of claims 1 to 11 and 20 to 26.

28. A pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of claims 1 to 11 and 16, for use in a method of treating or preventing an IL2Rγ-mediated disease or condition in a subject requiring such treatment, wherein the IL2Rγ-mediated disease or condition is graft-versus-host disease or an autoimmune disease.

29. The pharmaceutical composition according to claim 28, wherein the autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, or type 1 diabetes.

30. The pharmaceutical composition according to claim 28 or 29, wherein the antibody or its antigen-binding fragment is administered subcutaneously, intravenously, or intramuscularly to the body of a target by injection.

31. A pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of claims 1 to 11 and 16, for use in a method of treating or preventing an IL2Rγ-mediated disease or condition in a subject requiring such treatment, wherein the IL2Rγ-mediated disease or condition is β-islet cell graft rejection, skin graft rejection, heart graft rejection, lung graft rejection, kidney graft rejection, or liver graft rejection.

32. The pharmaceutical composition according to claim 31, wherein the antibody or its antigen-binding fragment is administered subcutaneously, intravenously, or intramuscularly to the body of a target by injection.

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