Anti-CD122 antibodies and uses thereof

Abstract

Provided herein are antibody molecules that specifically bind to CD122 and its antigen-binding portion, as well as related compositions, nucleic acid molecules, vectors, and host cells. Medical uses of such antibody molecules are also disclosed herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 279,762, filed November 16, 2021, and U.S. Provisional Patent Application No. 63 / 174,772, filed April 14, 2021, each of which is incorporated by reference in its entirety herein for all purposes.

[0002] Description of electronically submitted text files The contents of the text file submitted electronically herewith are incorporated herein by reference in their entirety: Copy of Sequence Listing in Computer Readable Format (Filename: VLRS_001_02WO_SeqList_ST25.txt, Data Recorded Date: April 13, 2022, File Size: Approximately 87,964 bytes).

[0003] The present disclosure relates to therapeutic antibody molecules and their medical uses. [Background technology]

[0004] CD122 is a cell surface receptor that is a member of the immunoglobulin superfamily and is expressed primarily on natural killer (NK) and T cells. CD122 has been proposed as a target for a variety of conditions mediated by any of these immune cell types, including type 1 diabetes (T1D), celiac disease, leukemia, vitiligo, etc. There is a need for therapeutic agents that target such immune-mediated diseases. In particular, vitiligo has no systemic treatment options and no disease-modifying medical treatments approved by the U.S. Food and Drug Administration. Summary of the Invention

[0005] Provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a heavy chain variable (VH) region, and a light chain variable (VL) region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0006] Provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:9.

[0007] In some embodiments, the antibody or antigen-binding portion is humanized or chimeric.

[0008] In some embodiments, the VH region, the VL region, or both the VH and VL regions comprise one or more human framework region amino acid sequences. In some embodiments, the VH region, the VL region, or both the VH and VL regions comprise a human variable region framework scaffold amino acid sequence into which the CDR amino acid sequences are inserted. In some embodiments, the VH region comprises an IGHV3-23 human germline scaffold amino acid sequence into which the HCDR1, HCDR2, and HCDR3 amino acid sequences are inserted. In some embodiments, the VL region comprises an IGKV1-33 human germline scaffold amino acid sequence into which the LCDR1, LCDR2, and LCDR3 amino acid sequences are inserted.

[0009] In some embodiments, the anti-CD122 antibody comprises an immunoglobulin constant region. In some embodiments, the immunoglobulin constant region is IgG, IgE, IgM, IgD, IgA, or IgY. In some embodiments, the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2. In some embodiments, the immunoglobulin constant region is immunologically inert. In some embodiments, the immunoglobulin constant region is a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG1 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A, or a wild-type human IgG2 constant region, numbering according to the EU index in Kabat. In some embodiments, the immunoglobulin constant region comprises any one of SEQ ID NOs: 32-38.

[0010] In some embodiments, the antibody or antigen binding portion is a Fab, Fab', F(ab')2, Fv, scFv, maxibody, minibody, diabody, triabody, tetrabody, or bis-scFv. In some embodiments, the antibody is monoclonal. In some embodiments, the antibody is a tetrameric antibody, a tetravalent antibody, or a multispecific antibody. In some embodiments, the antibody is a bispecific antibody that specifically binds to a first antigen and a second antigen, wherein the first antigen is CD122 and the second antigen is not CD122.

[0011] Provided herein is an immunoconjugate comprising an antibody or antigen-binding portion disclosed herein that is linked to a therapeutic agent. In some embodiments, the therapeutic agent is a cytotoxin, a radioisotope, a chemotherapeutic agent, an immunomodulatory agent, a cytostatic enzyme, a cytolytic enzyme, a therapeutic nucleic acid, an antiangiogenic agent, an antiproliferative agent, or a proapoptotic agent.

[0012] Provided herein are pharmaceutical compositions comprising the antibodies, antigen-binding portions, or immunoconjugates disclosed herein and a pharma- ceutically acceptable carrier, diluent, or excipient.

[0013] Provided herein are nucleic acid molecules encoding (a) the amino acid sequence of the VH region, (b) the amino acid sequence of the VL region, or (c) both the amino acid sequences of the VH and VL regions of an antibody or antigen-binding portion disclosed herein. Provided herein are expression vectors comprising the nucleic acid molecules disclosed herein. Provided herein are recombinant host cells comprising the nucleic acid molecules or expression vectors disclosed herein.

[0014] Provided herein is a method for producing an anti-CD122 antibody or antigen-binding portion thereof, comprising culturing a recombinant host cell comprising an expression vector disclosed herein under conditions in which the nucleic acid molecule is expressed, thereby producing the antibody or antigen-binding portion, and isolating the antibody or antigen-binding portion from the host cell or culture medium.

[0015] Provided herein are methods of suppressing an immune response in a subject, comprising administering to the subject a therapeutically effective amount of an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein. In some embodiments, the immune response is mediated by CD122.

[0016] Provided herein is a method for treating or preventing disease in a subject, comprising administering to said subject an effective amount of the antibody, antigen-binding portion, immunoconjugate or pharmaceutical composition disclosed herein.In some embodiments, the disease is inflammatory disease or autoimmune disease.In some embodiments, the disease is vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis or rheumatoid arthritis.

[0017] Provided herein is a method for inhibiting IL-15-induced T cell migration from the skin, comprising contacting the skin with a therapeutically effective amount of an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein.

[0018] Provided herein is an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein for use as a medicament. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 shows a diagram for the in vivo pharmacological analysis of anti-CD122 IgG. [Diagram 2] A-D show data from the specificity analysis of Villmab-1 (MIKβ1) anti-CD122 IgG. Proteomic specificity profiling of Villmab-1 using Retrogenix technology. (A) ZS control expression. (B) Villmab-1 probe. (C) Rituximab probe. (D) No primary antibody. [Figure 3A]Data from flow cytometry analysis of Villmab-1 (MIKβ1) anti-CD122 IgG in target transfected cells are shown. Binding of Villmab-1. [Figure 3B] Data from flow cytometry analysis of Villmab-1 (MIKβ1) anti-CD122 IgG in target transfected cells are shown, without primary antibody. [Figure 3C] Data from flow cytometry analysis of Villmab-1 (MIKβ1) anti-CD122 IgG in target transfected cells are shown. Rituximab binding. [Figure 4] Data from anti-CD122 IgG Alphascreen epitope competition are shown. New clones were screened in the Alphascreen assay to investigate competition with the Villmab-1 binding epitope on human CD122. [Diagram 5] 1 shows a bar graph depicting the polyreactivity scores for novel anti-CD122 IgG. The ability of the antibody to non-specifically bind to DNA and insulin was investigated. [Figure 6A] Shown are data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. 06F11. [Figure 6B] 07C07 shows data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. [Figure 6C] Shown are data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. 07D06. [Figure 6D] 07E09. Data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. [Figure 6E] 07D07 shows data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. [Figure 6F] Shown are data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. 06D12. [Figure 6G] Data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells are shown. Isotype control IgG. [Figure 7A] 06F11. Data from analysis of novel anti-CD122 IgG and Villmab-1 in the M07e cell IL-15 proliferation assay. [Figure 7B] 07C07. Data from analysis of novel anti-CD122 IgG and Villmab-1 in the M07e cell IL-15 proliferation assay. [Figure 7C] 07D06. Data from analysis of novel anti-CD122 IgG and Villmab-1 in the M07e cell IL-15 proliferation assay. [Figure 7D] 07E09. Data from analysis of novel anti-CD122 IgG and Villmab-1 in the M07e cell IL-15 proliferation assay. [Figure 7E] 07D07 shows data from analysis of novel anti-CD122 IgG and Villmab-1 in the M07e cell IL-15 proliferation assay. [Figure 7F] 06D12. Data from analysis of novel anti-CD122 IgG and Villmab-1 in an M07e cell IL-15 proliferation assay. [Figure 8A] Data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model are shown. Human CD8+ T cells before analysis. [Figure 8B] Data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model are shown. Human NK cells before analysis. [Figure 8C]Shown are data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model. Human CD8+ T cells after 1 week. [Figure 8D] Shown are data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model. Human NK cells after 1 week. [Figure 8E] Shown are data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model. Human CD8+ T cells after 3 weeks. [Figure 8F] Shown are data from in vivo analysis of novel anti-CD122 IgG and Villmab-1 in the hIL-15 NSG mouse model. Human NK cells after 3 weeks. [Figure 9A] Shown are data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. 06F11. [Figure 9B] 07C07 shows data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells. [Figure 9C] Data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells are shown. Isotype control IgG. [Figure 9D] Data from flow cytometry analysis of novel anti-CD122 IgG and Villmab-1 in target transfected cells are shown. Rituximab. [Figure 10A] Data from the specificity analysis of Villmab-1 (MIKβ1) anti-CD122 novel clones in Fab and IgG formats are shown. Using BIACORE® technology, the specificity profiling of Fab against human neudecin protein was measured as Rmax (maximum specific binding response value). [Figure 10B]Figure 1 shows data from the specificity analysis of Villmab-1 (MIKβ1) anti-CD122 novel clone in Fab and IgG format. Using BIACORE® technology, the specificity profiling of Fab against CILP2 protein was measured as Rmax (maximum specific binding response value). [Figure 10C] Specificity profiling of IgG against human BCAM protein was measured as OD450nm using ELISA. [Figure 11A] 1 shows data from sequence analysis of novel clone variable domain sequences of VillMab-1 (MIKβ1) anti-CD122. VH sequences. Sequences are as follows: VillMAB-1: SEQ ID NO:22, MAB05: SEQ ID NO:1, MAB06: SEQ ID NO:1, MAB14: SEQ ID NO:52, MAB15: SEQ ID NO:52, MAB17: SEQ ID NO:53, MAB18: SEQ ID NO:53. CDRs are bolded and underlined. Residues that differ from the VillMab-1 sequence are highlighted in grey boxes. Unique residues found only in clones from this analysis that do not bind BCAM, CILP2, or neudecin are boxed in black. [Figure 11B] 1 shows data from sequence analysis of novel clone variable domain sequences of VillMab-1 (MIKβ1) anti-CD122. VL sequence. Sequences are as follows: VillMAB-1: SEQ ID NO:28, MAB05: SEQ ID NO:9, MAB06: SEQ ID NO:17, MAB14: SEQ ID NO:9, MAB15: SEQ ID NO:17, MAB17: SEQ ID NO:9, MAB18: SEQ ID NO:17. CDRs are bolded and underlined. Residues that differ from the VillMab-1 sequence are highlighted in grey boxes. Unique residues found only in clones from this analysis that do not bind BCAM, CILP2, or neudecin are boxed in black. [Figure 12A] 1 shows data from analysis of a novel anti-CD122 IgG and Villmab-1 in a primary NK cell IL-15 proliferation assay. MAB05. [Figure 12B]1 shows data from analysis of a novel anti-CD122 IgG and Villmab-1 in a primary NK cell IL-15 proliferation assay. MAB06. [Figure 13] AB show data from an analysis of the effect of MAB05 and MAB06 on IL15-induced accumulation of T cells migrating from skin biopsies in a human skin biopsy culture assay. A shows the effect on the number of CD8+ T cells. B shows the effect on the number of CD4+ T cells. [Figure 14A] 1 shows data from an analysis of MAB05 and MAB06 concentration-dependent antagonism of IL15-induced accumulation of T cells migrating from skin biopsies in a human skin biopsy culture assay. Effects on CD8+ T cells and associated IC50s are shown. [Figure 14B] 1 shows data from an analysis of MAB05 and MAB06 concentration-dependent antagonism of IL15-induced accumulation of T cells migrating from skin biopsies in a human skin biopsy culture assay. Effects on CD4+ T cells and associated IC50s are shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Provided herein are anti-CD122 antibodies, and therapeutic uses of such antibodies. The antibodies disclosed herein are antagonistic, well expressed, biophysically stable, highly soluble, and have maximal identity to the preferred human germline.

[0021] CD122 (also known as IL2RB, IL-2Rβ, IL15RB, P70-75, interleukin 2 receptor subunit β, and IMD63) is a type I transmembrane glycoprotein and a member of the Ig superfamily. CD122 is a shared subunit of the interleukin-15 (IL-15) and interleukin-2 (IL-2) receptors. CD122 is expressed by NK cells and a subset of T cells. IL-15 signaling has been implicated in the pathogenesis of human vitiligo. Targeting CD122 or blocking IL-15 signaling appears to be beneficial in improving symptoms in mouse models of other immune-mediated diseases, such as diabetes, psoriasis, multiple sclerosis, and alopecia areata, as well as rheumatoid arthritis and celiac disease. Developing effective antagonistic anti-CD122 antibodies would be valuable in the treatment of immune-mediated diseases.

[0022] U.S. Patent No. 5,585,089, the entirety of which is incorporated herein by reference, describes the preparation of an antagonistic murine anti-CD122 IgG molecule, called "MIKβ1", as well as humanized forms of MIKβ1. These humanized forms of MIKβ1 were produced using classical humanization techniques, i.e., by grafting mouse CDRs defined by Kabat onto human heavy and light chain framework sequences (with some of the human framework residues optionally backmutated to correspondingly located MIKβ1 mouse residues). A partially humanized version of MIKβ1 (see Table 20) did not show efficacy in phase IIa clinical trials for T-cell large granular lymphocytic (T-LGL) leukemia and human T-cell lymphotropic virus 1 (HTLV-1)-associated myelopathy / tropical spastic paraparesis (HAM / TSP). This antibody has a number of disadvantages, including the fact that it is partially humanized, has off-target binding that may affect pharmacokinetics and biodistribution, and uses an IgG1 isotype that leads to the risk of unwanted antibody-dependent cellular cytotoxicity / antibody-dependent cellular phagocytosis in non-disease mediating cells. These characteristics make this antibody a suboptimal candidate for further testing as a targeted therapeutic in human immune-mediated diseases.

[0023] In contrast, the anti-CD122 antibodies provided herein exhibit advantages described herein that make them useful as therapeutic agents for human immune-mediated diseases and disorders.

[0024] antibody Provided herein are antibodies that specifically bind to CD122, and antigen-binding portions thereof. The anti-CD122 antibodies provided herein have several advantages over the murine anti-CD122 antibody MIKβ1, and its humanized versions, disclosed in US 5,585,089. The anti-CD122 antibodies provided herein have been selected to have improved efficacy in blocking IL-15 signaling by CD122. Importantly, these antibodies have dramatically improved specificity of CD122 binding compared to MIKβ1 by eliminating off-target binding to the human receptors BCAM (AU, CD239, LU, MSK19, basal adhesion molecule (Lutheran blood group), neudecin (also known as NENF), and CILP2 (also known as cartilage intermediate layer protein 2).

[0025] The antibodies and antigen binding portions disclosed herein specifically bind human CD122. In some embodiments, the antibodies and antigen binding portions may cross-react with CD122 from species other than human, such as Macaca fascicularis CD122 and / or Macaca mulatta CD122. In some embodiments, the antibodies may be specific for only human CD122 and may not exhibit non-human cross-reactivity. Exemplary human, cynomolgus, and rhesus CD122 amino acid sequences are shown in Table 16.

[0026] The term "antibody" refers broadly to an immunoglobulin (Ig) molecule, which generally comprises four polypeptide chains, two heavy (H) chains and two light (L) chains, or any functional fragment, mutant, variant, or derivative thereof that retains the essential target binding characteristics of an Ig molecule. Such mutant, variant, or derivative antibody formats are known in the art.

[0027] In full-length antibodies, each heavy chain comprises a heavy chain variable region (abbreviated herein as "VH region") and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as "VL region") and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FRs). Each VH and VL domain is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The definition of CDR used in this application is that of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, 5 th ed. Bethesda, MD: Public Health Service, National Institutes of Health (1991)).

[0028] The term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain. The "Fc region" may be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain may vary, the human IgG heavy chain Fc region is usually defined as extending from an amino acid residue at position Cys226, or from Pro230, to its carboxyl terminus. The numbering of residues in the Fc region is according to the EU index in Kabat. The Fc region of an immunoglobulin gene contains two constant domains, CH2 and CH3. The Fc region can exist in dimeric or monomeric form. The Fc region binds to various cellular receptors, such as Fc receptors, and other immune molecules, such as complement proteins.

[0029] Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY) and class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2) or subclass. IgG, IgD, and IgE antibodies generally contain two identical heavy chains and two identical light chains and two antigen-combining domains, each composed of a VH and a VL. IgA antibodies are generally composed of two monomers, each composed of two heavy chains and two light chains (as for IgG, IgD, and IgE antibodies), thus an IgA molecule has four antigen-binding domains, each again composed of a VH and a VL. Certain IgA antibodies are monomeric in that they are composed of two heavy chains and two light chains. Secreted IgM antibodies are generally composed of five monomers, each composed of two heavy chains and two light chains (as for IgG and IgE antibodies). Thus, an IgM molecule has 10 antigen-binding domains, each again composed of a VH and a VL. The cell surface form of IgM has a two heavy chain / two light chain structure, similar to IgG, IgD, and IgE antibodies.

[0030] As used herein, the term "antigen-binding portion" or "antigen-binding fragment" of an antibody (or "antibody portion" or "antibody fragment") means one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., CD122). It has been shown that the antigen-binding function of an antibody can be performed by a portion or a fragment of a full-length antibody. Examples of binding moieties encompassed by the term "antigen-binding portion" of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (v) a dAb (domain antibody) fragment comprising a single variable domain (Ward et al., (1989) Nature 341:544-546; WO 90 / 05144 A1, each of which is incorporated herein by reference in its entirety); and (vi) an isolated complementarity determining region (CDR). The present disclosure also encompasses Fab' fragments. Fab' fragments are formed by reduction of F(ab')2 fragments. Fab' is derived from F(ab')2, therefore Fab' may contain a small portion of Fc. Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be recombinantly linked by a synthetic linker that allows for the creation of a single protein chain (single-chain Fv (scFv)) in which the pair of VL and VH domains forms a monovalent molecule. See, e.g., Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883. Such single-chain antibodies are also intended to be encompassed by the term "antigen-binding portion" of an antibody. In some embodiments, scFv molecules can be incorporated into fusion proteins. In some embodiments, single-chain camelid antibodies are provided herein. In some embodiments, shark heavy chain antibodies (V-NARs) are provided herein.See English et al. (2020) Antibody Therapeutics, 3(1):1-9. Examples of antigen-binding moieties are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790 pp.). In some embodiments, single domain antibodies are provided herein. Generally, as used herein, the term "antibody" encompasses "antibody portions." Antibody portions generally retain the antigen-binding properties of full-length antibodies.

[0031] The antibodies and antibody portions provided herein may be in a multispecific (e.g., bispecific or trispecific) format. Such multispecific molecules specifically bind to two or more different molecular targets or epitopes. In some embodiments, the antibody or antigen-binding portion is a bispecific molecule that specifically binds to a first antigen and a second antigen, where the first antigen is CD122 and the second antigen is not CD122. In some embodiments, the antibody or antigen-binding portion is a diabody. Diabodies are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain, but the domains are forced to pair with complementary regions on another chain, creating two antigen-binding sites, by using a linker that is too short to allow pairing between the two domains on the same chain (see, e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak et al. (1994) Structure 2:1121-1123). In some embodiments, the antibody or antigen-binding portion is a triabody, tetrabody, bis-scFv, or tandem-scFv. In some embodiments, the antibody or antigen-binding portion is a dual affinity retargeting protein.

[0032] In some embodiments, the anti-CD122 antigen binding portion disclosed herein is a Fab, Fab', F(ab')2, Fv, scFv, maxibody, minibody, diabody, triabody, tetrabody, or bis-scFv.

[0033] As used herein, the terms "immunological binding" and "immunological binding properties" refer to the type of non-covalent interactions that occur between an immunoglobulin molecule (e.g., an antibody or an antigen-binding portion thereof) and an antigen for which the immunoglobulin is specific. The strength or affinity of an immunological binding interaction is determined by the dissociation constant (K d ) and smaller K d represents a greater affinity. The immunological binding properties of a selected polypeptide can be quantified using methods well known in the art. One such method involves measuring the rates of antigen-binding site / antigen complex formation and dissociation, which depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that affect the rate equally in both directions. Thus, the "on-rate constant" (K on ) and "off rate constant" (K off Both the K and the KD can be determined by calculating the concentration and the actual rates of association and dissociation (see Malmqvist, Nature 361:186-187 (1993)). off / K on The ratio of d (See Davies et al. (1990) Annual Rev Biochem 59:439-473). The antibodies or antigen-binding portions provided herein have an equilibrium binding constant (K d An antibody is said to specifically bind to CD122 when its K is ≦10 μM, preferably ≦10 nM, more preferably ≦10 nM, and most preferably ≦100 pM to about 1 pM. dOne method of measuring is by using surface plasmon resonance (SPR), usually a biosensor system such as a Biacore® system.

[0034] In some embodiments, the anti-CD122 antibodies or antigen-binding portions provided herein are monovalent or bivalent and comprise a single chain or two chains. Functionally, the binding affinity of the antibody or antigen-binding portion is greater than or equal to 10 -5 M~10 -12 For example, the binding affinity of an antibody or antigen-binding portion may be in the range of 10 -6 M~10 -12 M, 10 -7 M~10 -12 M, 10 -8 M~10 -12 M, 10 -9 M~10 -12 M, 10 -5 M~10 -11 M, 10 -6 M~10 -11 M, 10 -7 M~10 -11 M, 10 -8 M~10 -11 M, 10 -9 M~10 -11 M, 10 -10 M~10 -11 M, 10 -5 M~10 -10 M, 10 -6 M~10 -10 M, 10 -7 M~10 -10 M, 10 -8 M~10 -10 M, 10 -9 M~10 -10 M, 10 -5 M~10 -9 M, 10 -6 M~10 -9 M, 10 -7 M~10 -9 M, 10 -8 M~10 -9 M, 10 -5 M~10 -8 M, 10 -6 M~10 -8 M, 10 -7 M~10-8 M, 10 -5 M~10 -7 M, 10 -6 M~10 -7 M, or 10 -5 M~10 -6 It's M.

[0035] Provided herein are anti-CD122 antibodies or antigen-binding portions thereof, which cross-compete with antibody MAB05 or MAB06, or with an antibody comprising one or more amino acid sequences of antibody MAB05 or MAB06, for binding to CD122 (see Tables 18 and 19).

[0036] The terms "cross-compete", "cross-compete", "cross-block", "cross-blocked" and "cross-blocking" are used interchangeably herein to refer to the ability of an antibody or antigen-binding portion thereof to interfere with direct binding to a target CD122 (e.g., human CD122) or indirect binding by allosteric modulation of the anti-CD122 antibody of the present disclosure. The degree to which an antibody or portion thereof can interfere with the binding of another antibody or portion thereof to a target, and therefore whether the antibody or portion thereof can be said to be capable of cross-blocking or cross-competing, can be measured using a competitive binding assay. One example of a binding competition assay is homogeneous time-resolved fluorometry (HTRF). One particularly suitable quantitative cross-competition assay uses a FACS or Alphascreen-based approach to measure the competition between a labeled (e.g., His-tagged, biotinylated, or radiolabeled) antibody or portion thereof and another antibody or portion thereof in terms of binding to a target. Generally, a cross-competing antibody or portion thereof is one that binds to a target in a cross-competition assay such that, for example, in the assay and in the presence of a second antibody or portion thereof, the reported displacement of an immunoglobulin single variable domain or polypeptide according to the invention is up to 100% (e.g., in a FACS-based competition assay) of the theoretical maximum displacement (e.g., displacement by a cold (e.g., unlabeled) antibody or fragment thereof that needs to be cross-blocked) by a potential cross-blocking antibody or fragment thereof present in a given amount. In some embodiments, the cross-competing antibody or portion thereof has a reported displacement of between 10% and 100%, or between 50% and 100%.

[0037] Provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, which cross-competes for binding to CD122 with an antibody comprising a VH region and a VL region, and wherein (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0038] An anti-CD122 antibody or antigen-binding portion thereof, which cross-competes with an antibody or antigen-binding portion thereof comprising a set of CDRs disclosed herein for binding to CD122, and (a) specifically binds to (i) human CD122, and (ii) cynomolgus and / or rhesus monkey CD122, and (b) binds to human CD122, such as primary NK cells, when stimulated with human IL-15. + Provided herein are antibodies or antigen-binding portions thereof that (c) antagonize cell proliferation with an EC50 of less than 14 nM, (d) bind to rhesus CD122 with a KD of less than 10 nM, (e) bind to a functionally identical epitope on cynomolgus and / or rhesus CD122, and human CD122, and / or (f) exhibit reduced or no binding to BCAM compared to an anti-CD122 antibody comprising the variable domain sequence of antibody MIKβ1. In some embodiments, the KD of the antibody or antigen-binding portion is greater than or equal to 10 nM. d Values ​​can be measured by BIACORE® analysis. In some embodiments, the EC50 value of an antibody or antigen-binding portion can be measured by flow cytometry staining of cells expressing CD122 (e.g., CHO cells, HEK cells, M07e cells, NK cells, T cells).

[0039] In some embodiments, the anti-CD122 antibodies or antigen binding portions provided herein have reduced immunogenicity. In some embodiments, the antibodies or antigen binding portions exhibit reduced immunogenicity compared to an anti-CD122 antibody that comprises an HCDR1 of SYGVH (SEQ ID NO: 24), an HCDR2 of VIWSGGSTDYNAAFIS (SEQ ID NO: 5), an HCDR3 of AGDYNYDGFAY (SEQ ID NO: 27), an LCDR1 of SGSSSVSFMY (SEQ ID NO: 30), an LCDR2 of DTSNLAS (SEQ ID NO: 13), and an LCDR3 of QQWSTYPLT (SEQ ID NO: 15). In some examples, the immunogenicity risk of an antibody or antigen binding portion can be determined in silico by identifying the location of T cell epitopes in the antibody or portion (e.g., in the variable region or portion of the antibody).

[0040] For example, T cell epitopes in antibody or antigen binding sites can be identified using iTope™. iTope™ can be used to analyze VL and VH region sequences of peptides that bind with promiscuous high affinity to human MHC class II. Promiscuous high affinity MHC class II binding peptides are believed to correlate with the presence of T cell epitopes, which are high-risk indicators for clinical immunogenicity of drug proteins. iTope™ software predicts favorable interactions between peptide amino acid side chains and specific binding pockets (particularly pocket positions p1, p4, p6, p7, and p9) in the open end binding groove of 34 human MHC class II alleles. These alleles represent the most common HLA-DR alleles found worldwide, without weighting towards those most commonly found in any particular ethnic group. Twenty of the alleles contain the "open" p1 configuration, and 14 contain the "closed" configuration, with glycine at position 83 replaced by valine. Location of key binding residues is achieved by in silico generation of nonamer peptides that overlap with eight amino acids spanning the test protein sequence. This process successfully distinguishes, with great precision, between peptides that bind to MHC class II molecules and those that do not.

[0041] T cell epitopes in an antibody or antigen binding site can be identified by analyzing the VL and VH region sequences using TCED™ (T Cell Epitope Database™), which searches other protein sequences for matches to T cell epitopes previously identified by in vitro human T cell epitope mapping analysis. TCED™ is used to search any test sequence against a large (>10,000 peptides) database of peptides derived from unrelated protein and antibody sequences.

[0042] In some embodiments, an anti-CD122 antibody or antigen-binding portion may exhibit low immunogenicity because the antibody or portion has a small number of one or more of the following peptides in its sequence: high affinity foreign ("HAF" - high immunogenicity risk), low affinity foreign ("LAF" - low immunogenicity risk), and / or TCED+ (epitopes previously identified in the TCED™ database).

[0043] In some embodiments, an anti-CD122 antibody or antigen binding portion may have high germline epitope (GE) content in its sequence. In some examples, an anti-CD122 antibody or antigen binding portion has 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 (or more than 20) germline epitopes in its sequence (e.g., in the VL and / or VH region sequence). A germline epitope can be defined as a human germline peptide sequence with high MHC class II binding affinity. A nonamer peptide of a germline epitope is likely not immunogenic due to T cell tolerance, as demonstrated by previous studies using a wide range of germline peptides. Importantly, such germline v-domain epitopes (further supported by similar sequences in human antibody constant regions) compete for MHC class II occupancy at the membrane of antigen-presenting cells, reducing the risk of sufficient foreign peptide presentation to achieve the "activation threshold" required for T cell stimulation. High GE content can therefore be a beneficial quality in the clinical development of antibody therapeutics, resulting in low immunogenicity. In some examples, anti-CD122 antibodies or antigen-binding portions contain human germline peptide sequences (e.g., germline epitopes) with high MHC class II binding affinity in LCDR2.

[0044] In certain embodiments, an anti-CD122 antibody or antigen binding portion may have a reduced number of HAF, LAF, and / or TCED+ epitopes found in the frameworks of both the heavy and light chain variable regions compared to an anti-CD122 antibody comprising the variable domain sequence of antibody MIKβ1. In some embodiments, the HAF, LAF, and / or TCED+ epitopes are absent in the VL and / or VH region sequences of the anti-CD122 antibody or antigen binding portion.

[0045] In some embodiments, the anti-CD122 antibody or antigen-binding portion thereof does not comprise one or more of the MIKβ1 mouse / humanized antibody amino acid sequences shown in Table 20. In some embodiments, the anti-CD122 antibody or antigen-binding portion thereof does not comprise an HCDR1 comprising SEQ ID NO:24, an HCDR3 comprising SEQ ID NO:27, and / or an LCDR1 comprising SEQ ID NO:30. Table 1 shows the amino acid sequences of the MIKβ1 mouse anti-CD122 antibody variable regions with the CDRs highlighted as defined herein ("Kabat" scheme). The term "MIKβ1-IgG1 (humanized)" refers to an anti-CD122 antibody comprising the variable heavy chain region sequence labeled "CD122-VH1" and the variable light chain region sequence labeled "CD122-VL1" in Table 2, and a human IgG1 constant region.

[0046] The antibodies disclosed herein are anti-CD122 antagonist antibodies. As used herein, "antagonist" or "anti-CD122 antagonist antibody" (also interchangeably referred to as "anti-CD122 antibody") refers to an antibody capable of binding to CD122 and inhibiting CD122 biological activity and / or downstream pathway(s) mediated by CD122 signaling. Anti-CD122 antagonist antibodies encompass antibodies capable of blocking, antagonizing, inhibiting, or reducing (including significantly) CD122 biological activity, including downstream pathways mediated by CD122 signaling, e.g., receptor binding to CD122, and / or eliciting a cellular response to CD122. For purposes of this disclosure, the term "anti-CD122 antagonist antibody" will be expressly understood to encompass all terms, titles, and functional states and characteristics in which CD122 itself, and CD122 biological activity (including but not limited to the ability to inhibit activation of T cell anti-tumor cell activity), or the consequence of that activity or biological activity, is substantially abolished, reduced, or neutralized to a significant degree.

[0047] In some embodiments, the antibody molecule or antigen-binding portion thereof specifically binds to CD122 and does not bind (or does not specifically bind) to the membrane protein BCAM. In some embodiments, BCAM is a human protein. In some embodiments, BCAM is a rhesus protein. In some embodiments, the human BCAM protein comprises or consists of the amino acid sequence of SEQ ID NO:21, or an amino acid sequence that is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO:21. In one embodiment, the antibody molecule or antigen-binding portion thereof does not bind to BCAM. In some embodiments, the antibody molecule or antigen-binding portion thereof exhibits reduced binding to BCAM compared to binding to the membrane receptors as shown by antibody MIKβ1 or IgG1-MIKβ1 (humanized). In some cases, binding of the antibody or antigen-binding portion thereof to BCAM can be measured by ELISA or by flow cytometric analysis.

[0048] Further provided herein are anti-CD122 antibodies, or antigen-binding portions thereof, comprising one or more amino acid sequences of antibodies MAB06 or MAB05. The combinations of VH regions, VL regions, and CDR sequences forming these antibodies are shown in Tables 18 and 19. In some embodiments, the VH region sequence and / or the VL region sequence comprises a signal sequence (also known as a signal peptide) at the amino terminus.

[0049] Provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a heavy chain variable (VH) region, and a light chain variable (VL) region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0050] In some embodiments, there is provided an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody comprises a VH region comprising HCDR1, HCDR2, and HCDR3, and a VL region comprising LCDR1, LCDR2, and LCDR3, wherein (a) HCDR1 comprises the amino acid sequence GFTFSSY-X1-MS, where X1 is L, or any other amino acid (SEQ ID NO:39); and (b) HCDR2 comprises the amino acid sequence X1-A-X2-ISGGG-X3-X4-X5-YY-X6-DSVKG, where X1 is V, or a conservative substitution of V, X2 is T or N, X3 is A or S, X4 is E or N, X5 is T or K, and X6 is P or V. ] (SEQ ID NO: 40), and (c) HCDR3 comprises the amino acid sequence X1-X2-X3-X4-X5-DY, wherein X1 is Q or any other amino acid (e.g., T, L, M, or N), X2 is L or any other amino acid (e.g., G, K, M, Q, S, or V), X3 is Y or a conservative substitution of Y (e.g., H), X4 is Y or any other amino acid (e.g., A, D, F, G, M, E, I, K, S, or W), and X5 is F or any other amino acid (e.g., A, D, E, I, K, M, S, or W). ] (SEQ ID NO: 160); (d) LCDR1 comprises the amino acid sequence RASQSI-X1-X2-X3-X4-X5, where X1 is S or a conservative substitution of S, X2 is S or a conservative substitution of S, X3 is Y or a conservative substitution of Y, X4 is L or a conservative substitution of L, and X5 is N or T, or a conservative substitution of N or T. ] (SEQ ID NO: 161); (e) LCDR2 comprises the amino acid sequence X1-A-X2-SL-X3-X4, where X1 is A or T, or a conservative substitution of A or T, X2 is S or a conservative substitution of S, X3 is Q, or any other amino acid, and X4 is S, or any other amino acid. ] (sequence number 162), (f) LCDR3 comprises the amino acid sequence QQ-X1-YS-X2-P-X3-T, wherein X1 is S or any other amino acid, X2 is T or any other amino acid, and X3 is W or any other amino acid.] (SEQ ID NO: 163), or an antigen-binding portion thereof, is disclosed herein.

[0051] Provided herein is an anti-CD122 antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9.

[0052] Provided herein is an anti-CD122 antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region and a VL region, and the amino acid sequence of the VH region comprises SEQ ID NO:1, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:1.

[0053] Provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region and a VL region, and the amino acid sequence of the VL region comprises (a) an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:17 or SEQ ID NO:9, or (b) an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:17 or SEQ ID NO:9.

[0054] Also provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises an amino acid sequence of SEQ ID NO:1 or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:1, and the amino acid sequence of the VL region comprises an amino acid sequence of SEQ ID NO:17 or SEQ ID NO:9, or (b) the amino acid sequence of the VH region comprises an amino acid sequence of SEQ ID NO:1 or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:1, and the amino acid sequence of the VL region comprises an amino acid sequence of SEQ ID NO:9 or SEQ ID NO:17.

[0055] Provided herein is an anti-CD122 antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:17, and there are one, two, or three conservative amino acid substitutions in the VH region sequence, the VL region sequence, or both the VH region and VL region sequences, or (b) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:9, and there are one, two, or three conservative amino acid substitutions in the VH region sequence, the VL region sequence, or both the VH region and VL region sequences. In some embodiments, the conservative amino acid substitutions are made only in the FR sequences and not in the CDR sequences of the antibody or antigen-binding portion.

[0056] In some embodiments, the anti-CD122 antibodies or antigen-binding portions provided herein are monoclonal. The term "monoclonal antibody" (Mab) refers to an antibody or antigen-binding portion thereof derived from a single copy or clone, including, for example, any eukaryotic, prokaryotic, or phage clone, and does not refer to the method by which the antibody or antigen-binding portion thereof is produced. Preferably, monoclonal antibodies are present in a homogenous or substantially homogenous population.

[0057] In some embodiments, the antibodies or antigen-binding portions provided herein can be isolated.

[0058] In some embodiments, the anti-CD122 antibodies or antigen-binding portions provided herein are chimeric. The term "chimeric" is intended to mean an antibody molecule, or antigen-binding portion thereof, in which the variable domain sequences are derived from one species and at least one constant region sequence is derived from another species. For example, one or all of the variable domains of the light chain(s) and / or one or all of the variable domains of the heavy chain(s) of a mouse antibody (e.g., a mouse monoclonal antibody) can be linked to a human constant region, for example, but not limited to, an IgG1 or IgG4 human constant region, respectively. Examples of chimeric antibodies and suitable techniques for producing such antibodies are described in U.S. Patent Nos. 4,816,567, 4,975,369, and 4,816,397, each of which is incorporated by reference herein in its entirety. In some embodiments, an anti-CD122 antibody or antigen-binding portion provided herein comprises (a) an amino acid sequence of a VH region comprising SEQ ID NO:1, an amino acid sequence of a VL region comprising SEQ ID NO:17 and a human constant region, or (b) an amino acid sequence of a VH region comprising SEQ ID NO:1, an amino acid sequence of a VL region comprising SEQ ID NO:9 and a human constant region.

[0059] In some embodiments, the anti-CD122 antibodies or antigen-binding portions provided herein are humanized. The term "humanized" is intended to mean an antibody or antigen-binding portion thereof that has been engineered to include one or more human framework regions in the variable domains, along with non-human (e.g., mouse, rat, or hamster) CDRs of the heavy and / or light chains. In some embodiments, a humanized antibody comprises sequences that are fully human except for the CDRs. An anti-CD122 antibody molecule or antigen-binding portion thereof can include one or more human variable region framework scaffolds into which the CDRs are inserted. In some embodiments, the VH region, VL region, or both the VH region and the VL region of an anti-CD122 antibody or antigen-binding portion provided herein comprises one or more amino acid sequences of human framework regions. In some embodiments, a humanized antibody comprises sequences that are fully human except for the CDRs that are the CDRs of antibody MAB06 or MAB05. Examples of humanized antibodies, and techniques suitable for their production, can be found in Hwang et al., Methods 36:35, 2005; Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033, 1989; Jones et al., Nature, 321:522-25, 1986; Riechmann et al., Nature, 332:323-27, 1988; Verhoeyen et al., Science, 239:1534-36, 1988; Orlandi et al., al., Proc. Natl. Acad. Sci. USA, 86:3833-37, 1989, US5,225,539, US5,530,101, US5,585,089, US5,693,761, US5,693,762, US6,180,370, and WO90 / 07861, each of which is incorporated herein by reference in its entirety. When selecting FRs to flank CDRs, e.g., when humanizing or optimizing an antibody, FRs derived from antibodies containing CDR sequences in the same canonical class are preferred.

[0060] In some embodiments, the anti-CD122 antibodies or antigen-binding fragments provided herein do not necessarily have the maximal number of human germline substitutions at the corresponding murine CDR or other (such as framework) amino acid positions. As detailed in the Experimental Section below, a "maximally humanized" antibody molecule is not necessarily "maximally optimized" in terms of anti-CD122 binding properties and / or other desirable characteristics.

[0061] The present disclosure encompasses modifications to the amino acid sequence of an antibody molecule or antigen-binding portion thereof as defined herein. For example, the present disclosure includes antibody molecules, and corresponding antigen-binding portions thereof, that include functionally equivalent variable regions and CDRs that do not significantly affect the properties, as well as variants with improved or decreased activity and / or affinity. For example, the amino acid sequence can be mutated to obtain an antibody with a desired binding affinity for CD122. Amino- and / or carboxyl-terminal fusions, ranging in length from one residue to polypeptides containing 100 or more residues, as well as insertions including intrasequence insertions of single or multiple amino acid residues are contemplated. Examples of terminal insertions include antibody molecules with an N-terminal methionyl residue or fused to an epitope tag. Other insertion variants of antibody molecules include the fusion to the N- or C-terminus of the antibody of an enzyme or a polypeptide that increases the half-life of the antibody in the blood circulation.

[0062] In some embodiments, the anti-CD122 antibodies or antigen binding portions provided herein can include glycosylated and non-glycosylated polypeptides, as well as polypeptides that have undergone other post-translational modifications, such as glycosylation with different sugar groups, acetylation, and phosphorylation. The antibodies or antigen binding portions can be mutated to alter such post-translational modifications, for example, by adding, removing, or replacing one or more amino acid residues to create or remove a glycosylation site.

[0063] In some embodiments, the anti-CD122 antibodies or antigen-binding portions provided herein can be modified, for example, by amino acid substitution to remove potential proteolytic sites in the antibody or portion.

[0064] Also provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region, a VL region, and an entire human framework region sequence, and wherein (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0065] Also provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region, a VL region, and one or more human framework region sequences, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0066] In some embodiments, an anti-CD122 antibody, or antigen binding portion thereof, can comprise an IGHV3-23 human germline scaffold into which the corresponding HCDR sequences have been inserted. An anti-CD122 antibody, or antigen binding portion thereof, can comprise a VH region comprising an IGHV3-23 human germline scaffold amino acid sequence into which the corresponding set of HCDR1, HCDR2, and HCDR3 amino acid sequences have been inserted.

[0067] In some embodiments, an anti-CD122 antibody, or antigen binding portion thereof, can comprise an IGKV1-33 human germline scaffold into which a corresponding LCDR sequence has been inserted. An anti-CD122 antibody, or antigen binding portion thereof, can comprise a VL region comprising an IGKV1-33 human germline scaffold amino acid sequence into which a corresponding set of LCDR1, LCDR2, and LCDR3 amino acid sequences has been inserted.

[0068] In some embodiments, an anti-CD122 antibody or antigen binding portion thereof can comprise an IGHV3-23 human germline scaffold into which a corresponding HCDR sequence has been inserted, and an IGKV1-33 human germline scaffold into which a corresponding LCDR sequence has been inserted. An anti-CD122 antibody or antigen binding portion thereof can comprise a VH region comprising an IGHV3-23 human germline scaffold amino acid sequence into which a corresponding set of HCDR1, HCDR2, and HCDR3 amino acid sequences has been inserted, and a VL region comprising an IGKV1-33 human germline scaffold amino acid sequence into which a corresponding set of LCDR1, LCDR2, and LCDR3 amino acid sequences has been inserted. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 amino acid sequences may be the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 amino acid sequences of any one of the clones in Tables 18 or 19 (wherein all six CDR sequences are derived from the same clone).

[0069] In some embodiments, the anti-CD122 antibody or antigen-binding portion thereof comprises an immunoglobulin constant region. In some embodiments, the immunoglobulin constant region is IgG, IgE, IgM, IgD, IgA, or IgY. In some embodiments, the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2. In some embodiments, the immunoglobulin constant region is immunologically inert. In some embodiments, the immunoglobulin constant region comprises one or more mutations that reduce or prevent FcγR binding, antibody-dependent cellular cytotoxicity, and / or complement-dependent cytotoxicity. In some embodiments, the constant region of the immunoglobulin is a wild-type human IgG1 constant region, a wild-type human IgG2 constant region, a wild-type human IgG4 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, G237A, and P331S, or a human IgG4 constant region comprising the amino acid substitution S228P, where the numbering is according to the EU index in Kabat. In some embodiments, the positions of amino acid residues in the constant region of an immunoglobulin molecule are numbered according to the EU index in Kabat (Ward et al., 1995 Therap. Immunol. 2:77-94).

[0070] In some embodiments, the anti-CD122 antibody, or antigen-binding portion thereof, can comprise an immunoglobulin light chain constant region that is a kappa light chain constant region or a lambda light chain constant region.

[0071] In some embodiments, the anti-CD122 antibody can comprise an immunoglobulin constant region comprising any one of the amino acid sequences in Table 15. The Fc region sequences in Table 15 begin with the CH1 domain. In some embodiments, the anti-CD122 antibody can comprise an immunoglobulin constant region comprising the amino acid sequence of a human IgG4, a human IgG4(S228P), a human IgG2, a human IgG1, or a human IgG1 effector null Fc region. For example, a human IgG4(S228P) Fc region comprises the following substitution compared to a wild-type human IgG4 Fc region: S228P. For example, a human IgG1 effector null Fc region comprises the following substitutions compared to a wild-type human IgG1 Fc region: L234A, L235A, and G237A. In some embodiments, the immunoglobulin constant region can comprise an RDELT (SEQ ID NO: 39) motif, or a REEM (SEQ ID NO: 40) motif (underlined in Table 15). The REEM (SEQ ID NO: 40) allotype is found in a smaller proportion of the human population than the RDELT (SEQ ID NO: 39) allotype. In some embodiments, the anti-CD122 antibody can comprise an immunoglobulin constant region comprising any one of SEQ ID NOs: 32-38. In some embodiments, the anti-CD122 antibody can comprise the six CDR amino acid sequences of any one of the clones in Tables 18 or 19 and any one of the Fc region amino acid sequences in Table 15. In some embodiments, the anti-CD122 antibody can comprise an immunoglobulin heavy chain constant region comprising any one of the Fc region amino acid sequences in Table 15, and an immunoglobulin light chain constant region that is a kappa light chain constant region or a lambda light chain constant region.

[0072] In some embodiments, provided herein is an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody comprises a VH region, a VL region, and a heavy chain constant region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO: 3, an HCDR2 comprising SEQ ID NO: 5, and an HCDR3 comprising SEQ ID NO: 7, the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO: 18, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO: 15, and the heavy chain constant region comprises any one of SEQ ID NOs: 32 to 38, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO: 3, an HCDR2 comprising SEQ ID NO: 5, and an HCDR3 comprising SEQ ID NO: 7, the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO: 11, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO: 15, and the heavy chain constant region comprises any one of SEQ ID NOs: 32 to 38.

[0073] In some embodiments, there is provided an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody comprises a VH region, a VL region, and a heavy chain constant region, wherein (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region, a wild-type human IgG1 constant region, or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A. or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO: 3, an HCDR2 comprising SEQ ID NO: 5, and an HCDR3 comprising SEQ ID NO: 7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO: 11, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO: 15, and the heavy chain constant region comprises any one of SEQ ID NOs: 32 to 38, and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region, a wild-type human IgG1 constant region, or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A.

[0074] In some embodiments, provided herein is an anti-CD122 antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO: 1, the amino acid sequence of the VL region comprises or consists of SEQ ID NO: 17, and the heavy chain constant region comprises any one of SEQ ID NOs: 32-38, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO: 1, the amino acid sequence of the VL region comprises or consists of SEQ ID NO: 9, and the heavy chain constant region comprises any one of SEQ ID NOs: 32-38.

[0075] In some embodiments, the anti-CD122 antibody may be immune effector null. In some embodiments, the anti-CD122 antibody or antigen-binding portion thereof does not induce immune effector function, and optionally inhibits immune effector function. In some embodiments, the anti-CD122 antibody may lack measurable binding to human FcγRI, FcγRIIa, FcγRIIIa, and FcγRIIIb receptors, but may maintain binding to human FcγRIIb receptors, and may optionally maintain binding to human FcRn receptors. FcγRI, FcγRIIa, FcγRIIIa, and FcγRIIIb are examples of activating receptors. FcγRIIb is an example of an inhibitory receptor. FcRn is an example of a recycling receptor. In some embodiments, the binding affinity of the anti-CD122 antibody or antigen-binding portion thereof to human Fc receptors can be measured by BIACORE® analysis. In some embodiments, homogeneous time-resolved fluorometry (HTRF) can be used to investigate the binding of anti-CD122 antibodies to human Fc receptors. In one example of HTRF, human IgG1 (wild type) is labeled as a complete Fcγ receptor, and then antibodies containing recombinant Fc fragments are used in a titration competition. In some embodiments, CD122 positive cells can be mixed with human white blood cells and anti-CD122 antibodies, and cell killing by CDC, ADCC, and / or ADCP can be measured. In some embodiments, anti-CD122 antibodies that include the amino acid sequence of the Fc region of human IgG1 (see Table 15) are effector null. In some embodiments, anti-CD122 antibodies that include the amino acid sequence of the Fc region of human IgG1 (see Table 15) are not effector null.

[0076] Further provided herein is an immunoconjugate comprising an anti-CD122 antibody, or an antigen-binding portion thereof, linked to a therapeutic agent. In some embodiments, the therapeutic agent is a cytotoxin, a radioisotope, a chemotherapeutic agent, an immunomodulatory agent, a cytostatic enzyme, a cytolytic enzyme, a therapeutic nucleic acid, an antiangiogenic agent, an antiproliferative agent, or a proapoptotic agent.

[0077] Examples of suitable therapeutic agents include, but are not limited to, immunomodulatory agents, cytotoxins, radioisotopes, chemotherapeutic agents, antiangiogenic agents, antiproliferative agents, proapoptotic agents, and cytostatic and cytolytic enzymes (e.g., RNAses). Additional therapeutic agents include therapeutic nucleic acids, such as genes encoding immunomodulatory agents, antiangiogenic agents, antiproliferative agents, or proapoptotic agents. The terms for these agents are not mutually exclusive, and thus a therapeutic agent may be described using one or more of the above terms.

[0078] Examples of suitable therapeutic agents for use in immunoconjugates include, but are not limited to, JAK kinase inhibitors, taxanes, maytansines, CC-1065 and duocarmycins, calicheamicins and other enediynes, and auristatins. Other examples include antifolates, vinca alkaloids, and anthracyclines. Plant toxins, other bioactive proteins, enzymes (i.e., ADEPT), radioisotopes, photosensitizers can also be used in immunoconjugates. In addition, conjugates can be made using secondary carriers, such as liposomes or polymers, as cytotoxic agents. Suitable cytotoxins include agents that inhibit or prevent the function of cells and / or cause the destruction of cells. Exemplary cytotoxins include antibiotics, inhibitors of tubulin polymerization, alkylating agents that bind to and destroy DNA, and agents that disrupt protein synthesis or the function of essential cellular proteins, such as protein kinases, phosphatases, topoisomerases, enzymes, and cyclins.

[0079] Exemplary cytotoxins include doxorubicin, daunorubicin, idarubicin, aclarubicin, zorubicin, mitoxantrone, epirubicin, carubicin, nogalamycin, menogaril, pitarubicin, valrubicin, cytarabine, gemcitabine, trifluridine, ancitabine, enocitabine, azacytidine, doxifluridine, pentostatin, broxuhdine, capecitabine, cladofubin, decitabine, floxuhdine, fludarabine, gougerotin, puromycin, tegafur, thiazofuhn, adhamycin, in), cisplatin, carboplatin, cyclophosphamide, dacarbazine, vinblastine, vincristine, mitoxantrone, bleomycin, mechlorethamine, prednisone, procarbazine, methotrexate, fluorouracil, etoposide, taxol, taxol analogs, platins such as cis-platin and carboplatin, mitomycin, thiotepa, taxanes, vincristine, daunorubicin, epirubicin, actinomycin, anthramycin, azaserine, bleomycin, tamoxifen, idarubicin, dolastatins / auristatins, hemiasterlin, esperamicin, and maytansinoids.

[0080] Suitable immunomodulatory agents include anti-hormonal agents that block hormone action on tumors, and immunosuppressants that suppress cytokine production, downregulate self-antigen expression, or mask MHC antigens.

[0081] Pharmaceutical Compositions The anti-CD122 antibodies and antigen-binding portions (also referred to herein as "active compounds") provided herein can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically include an anti-CD122 antibody or antigen-binding portion (or an immunoconjugate comprising said antibody or portion) and a pharma- ceutically acceptable carrier, diluent, or excipient. Such materials should be non-toxic and should not interfere with the efficacy of the anti-CD122 antibody or antigen-binding fragment thereof. The precise nature of the carrier or other material will depend on the route of administration, which may be injection, bolus, infusion, or any other suitable route discussed below.

[0082] As used herein, the term "pharmaceutical acceptable" refers to a molecular entity or composition that does not generally produce allergic or other serious adverse reactions when administered using routes well known in the art. A molecular entity or composition that is approved by a federal or state government regulatory agency in the United States or is listed in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals, and more specifically in humans, is considered to be "pharmaceutical acceptable." As used herein, the term "pharmaceutical acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonicity agents, absorption delaying agents, and the like, that are compatible in pharmaceutical administration. Suitable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, a standard reference in this field, which is incorporated herein by reference. Some examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles, such as fixed oils, can also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, its use in the composition is contemplated. Supplementary active ingredients can also be incorporated into the composition. Pharmaceutically acceptable carriers, diluents, or excipients can be compounds or combinations of compounds that do not cause secondary reactions, for example, facilitate the administration of anti-CD122 antibody or its antigen-binding portion, increase its life span and / or effectiveness in the body, or increase its solubility in solution.

[0083] Provided herein is a pharmaceutical composition comprising: (i) an anti-CD122 antibody, or antigen-binding portion thereof, comprising a VH region and a VL region, wherein (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15; and (ii) a pharma- ceutically acceptable carrier, diluent, or excipient; or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15.

[0084] Provided herein is a pharmaceutical composition comprising: (i) an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9; and (ii) a pharma- ceutically acceptable carrier, diluent, or excipient.

[0085] The pharmaceutical compositions disclosed herein may be formulated to suit the intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous applications may include the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents, antibacterial agents such as benzyl alcohol or methylparabens, antioxidants such as ascorbic acid or sodium bisulfite, chelating agents such as ethylenediaminetetraacetic acid (EDTA), buffers such as acetates, citrates, or phosphates, and tonicity adjusting agents such as sodium chloride or dextrose. The pH can be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide. Parenteral preparations can be enclosed in glass or plastic ampoules, disposable syringes, or multiple dose vials.

[0086] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL® (BASF, Parsippany, NJ), or phosphate buffered saline (PBS). In all cases, the composition must be sterile and fluid for easy syringability. The composition must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it is preferable to include isotonic agents, for example, sugars, polyalcohols (such as mannitol, sorbitol, or sodium chloride) in the composition. Absorption of the injectable compositions can be delayed by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0087] Sterile injectable solution can be prepared by incorporating the active compound in the required amount in a suitable solvent with one or a combination of the above-listed components as required, and then sterilizing by filtration.Generally, dispersion is prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and other components required from the above-listed components.In the case of sterile powder for preparing sterile injectable solution, the preparation method is vacuum drying and freeze-drying, which produces a powder of the active ingredient plus any additional desired components from its previously sterile-filtered solution.

[0088] Oral compositions usually include an inert diluent or an edible carrier. The inert diluent or edible carrier can be enclosed in a gelatin capsule or compressed into a tablet. For the purpose of oral therapeutic administration, the active compound can be incorporated with an excipient and used in the form of a tablet, a troche, or a capsule. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, in which the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents and / or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primojel®, or corn starch; a lubricant such as magnesium stearate, a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin, or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0089] For administration by inhalation, the compounds can be delivered in the form of an aerosol spray from pressured container or insufflator which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

[0090] Systemic administration can also be by transmucosal or transdermal methods.For transmucosal or transdermal administration, a penetrant suitable for the barrier to be permeated is used in the formulation.Such penetrants are generally known in the art, and include, for example, for transmucosal administration, surfactants, bile salts, and fusidic acid derivatives.Transmucosal administration can be achieved by using nasal sprays or suppositories.For transdermal administration, the active compound is formulated into ointments, salves, gels, or creams generally known in the art.

[0091] The medications can also be prepared in the form of suppositories (eg, with conventional suppository bases such as cocoa butter or other glycerides) or retention enemas for rectal delivery.

[0092] In some embodiments, the active compound is prepared with a carrier that prevents the compound from being rapidly removed from the body, for example, a controlled release formulation, such as implants and microencapsulated delivery systems.Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used.The method of preparing such formulations will be clear to those skilled in the art.Materials can also be commercially obtained.Liposomal suspensions can also be used as pharmaceutically acceptable carriers.

[0093] It is particularly advantageous to prepare oral or parenteral compositions in dosage unit form, which is easy to administer and has a uniform dose.The term "unit dosage form" used herein refers to a physically separate unit suitable for a unit dosage for treating a subject, each unit containing a predetermined amount of active compound calculated to produce a desired therapeutic effect in association with the required pharmaceutical carrier.The specification of the unit dosage form of the present invention is determined and directly depends on the inherent characteristics of the active compound and the specific therapeutic effect to be achieved, as well as the inherent limitations of the technical field of preparing such active compound for treatment in individuals.

[0094] In some embodiments, the anti-CD122 antibody or antigen-binding portion thereof can be provided in lyophilized form for reconstitution prior to administration. For example, lyophilized antibody molecules can be reconstituted in sterile water and mixed with saline prior to administration to an individual.

[0095] The pharmaceutical compositions provided herein can be included in a container, pack, or dispenser together with instructions for administration.

[0096] Nucleic acid molecules, vectors, host cells, and methods for producing antibodies Provided herein are nucleic acid molecules (e.g., isolated nucleic acid molecules) that encode the amino acid sequence of an anti-CD122 antibody or anti-CD122 antigen binding portion described herein (or the amino acid sequence of (i) the VH region, (ii) the VL region, or (iii) both the VH and VL regions of the antibody or antigen binding portion). Further provided herein are nucleic acid molecules (e.g., isolated nucleic acid molecules) that encode (i) the heavy chain, (ii) the light chain, or (iii) both the heavy and light chains of an anti-CD122 antibody or anti-CD122 antigen binding portion described herein. In some embodiments, the nucleic acid molecule encoding the VH region, VL region, heavy chain, or light chain comprises a signal sequence. In some embodiments, the nucleic acid molecule encoding the VH region, VL region, heavy chain, or light chain does not comprise a signal sequence.

[0097] In some embodiments, the nucleic acid molecule encodes amino acid sequences of the VH and VL regions of an anti-CD122 antibody or antigen binding portion thereof, wherein (a) the amino acid sequence of the VH region comprises HCDR1 comprising SEQ ID NO:3, HCDR2 comprising SEQ ID NO:5, and HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises LCDR1 comprising SEQ ID NO:18, LCDR2 comprising SEQ ID NO:13, and LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises HCDR1 comprising SEQ ID NO:3, HCDR2 comprising SEQ ID NO:5, and HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises LCDR1 comprising SEQ ID NO:11, LCDR2 comprising SEQ ID NO:13, and LCDR3 comprising SEQ ID NO:15. In some embodiments, the nucleic acid molecule further encodes the amino acid sequences of human framework regions.

[0098] In some embodiments, the nucleic acid molecule encodes amino acid sequences of the VH region and the VL region of an anti-CD122 antibody, or antigen-binding portion thereof, wherein (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9.

[0099] Also provided herein is an expression vector comprising the nucleic acid molecule described herein. In certain vectors, the nucleic acid molecule is operably linked to one or more regulatory sequences suitable for expression of the nucleic acid segment in a host cell. In some cases, the expression vector comprises a sequence that mediates replication and includes one or more selectable markers. As used herein, "vector" refers to a construct capable of delivering, and preferably expressing, one or more gene(s) or sequence(s) of interest in a host cell. Examples of vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmids, cosmids or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, DNA or RNA expression vectors encapsulated in liposomes, and certain eukaryotic cells such as producer cells.

[0100] Provided herein is a recombinant host cell comprising an expression vector or nucleic acid molecule disclosed herein. A "host cell" includes an individual cell, cell line, or cell culture that can be or has been a recipient for a vector(s) for incorporating a polynucleotide insert. A host cell includes the progeny of a single host cell. The progeny may not necessarily be completely identical (in morphology or genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. An expression vector can be transfected into a host cell by standard techniques. Non-limiting examples include electroporation, calcium phosphate precipitation, DEAE-dextran transfection, and the like. In some embodiments, a recombinant host cell comprises a single vector or a single nucleic acid molecule encoding both the VH and VL regions of an anti-CD122 antibody or antigen-binding portion thereof. In some embodiments, the recombinant host cell comprises (i) a first vector or a first nucleic acid molecule encoding a VH region of an anti-CD122 antibody, or antigen-binding portion thereof, and (ii) a second vector or a second nucleic acid molecule encoding a VL region of an anti-CD122 antibody, or antigen-binding portion thereof.

[0101] The antibody molecules of the present invention, or antigen-binding portions thereof, can be produced using techniques well known in the art, such as recombinant techniques, phage display techniques, synthetic techniques, computational techniques, or a combination of such techniques or other techniques readily known in the art.

[0102] Further provided herein is a method of producing an anti-CD122 antibody or antigen-binding portion thereof, comprising culturing a recombinant host cell comprising an expression vector described herein under conditions in which the nucleic acid segment is expressed to produce the anti-CD122 antibody or antigen-binding portion thereof. The antibody or antigen-binding portion thereof can then be isolated from the host cell or culture medium. The anti-CD122 antibody or antigen-binding portion thereof can be produced by any of a variety of methods known to those of skill in the art. In certain embodiments, the anti-CD122 antibody and antigen-binding portion thereof can be produced recombinantly. For example, a nucleic acid sequence encoding one or more of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, or SEQ ID NO:18, or a portion thereof, can be introduced into a bacterial cell (e.g., E. coli, B. subtilis), or a eukaryotic cell (e.g., a yeast cell such as S. cerevisiae, or a mammalian cell such as a CHO cell line, various Cos cell lines, HeLa cells, HEK293 cells, various myeloma cell lines, or transformed B cells or hybridomas), or an in vitro translation system, and the translated polypeptide can be isolated. In some embodiments, the antibody light and heavy chain proteins are produced in the cell with signal sequences that are removed upon production of the mature anti-CD122 antibody or antigen-binding portion thereof.

[0103] One of skill in the art can determine whether an antibody or antigen-binding portion comprising a given polypeptide sequence binds to the CD122 protein without undue experimentation using standard methodologies, e.g., Western blot, ELISA, etc.

[0104] 1. A method for producing an antibody, or an antigen-binding portion thereof, that specifically binds to human CD122, and optionally also to cynomolgus and / or rhesus CD122, comprising: (1) grafting anti-CD122 CDRs derived from a non-human source onto a human v-domain framework to produce a humanized anti-CD122 antibody molecule, or an antigen-binding portion thereof; (2) generating a library of clones of humanized anti-CD122 antibody molecules, or antigen-binding portions thereof, that contain one or more mutations in the CDRs; (3) screening the library for binding to human CD122, and optionally also to cynomolgus and / or rhesus CD122; (4) selecting clones from screening step (3) that have binding specificity for human CD122, and optionally also for cynomolgus and / or rhesus CD122, but have reduced or no binding to human BCAM, human CILP2, or human neudecin; The above method is provided herein, comprising: (5) producing, from the clones selected in step (4), antibody molecules, or antigen-binding portions thereof, that specifically bind to human CD122, and optionally also to cynomolgus and / or rhesus monkey CD122.

[0105] The method may include the further step of generating additional clones based on the clones selected in step (4), e.g., based on further exploratory mutagenesis at specific positions in the CDRs of the clones selected in step (4), to improve humanization and / or minimize the content of human T-cell epitopes and / or improve manufacturing characteristics in the antibody molecules or antigen-binding portions thereof produced in step (5).

[0106] Use of antibodies Provided herein are methods and uses of the anti-CD122 antibodies, anti-CD122 antigen-binding portions, immunoconjugates, and pharmaceutical compositions described herein to provide a therapeutic benefit to a subject with an immune-mediated disease or disorder.

[0107] Provided herein is a method of suppressing an immune response in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein. Provided herein is a method for suppressing an immune response in a subject (e.g., an immune response mediated by CD122-positive cells), comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15. Provided herein are methods of suppressing an immune response in a subject, comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO: 9. In some embodiments, the immune response is mediated by CD122.

[0108] Provided herein is a method for inhibiting IL-15-induced T cell migration from the skin (e.g., human skin), comprising contacting the skin with a therapeutically effective amount of an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein. Provided herein is a method for inhibiting IL-15-induced T cell migration out of the skin (e.g., human skin), comprising contacting the skin with a therapeutically effective amount of an anti-CD122 antibody, or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15. Provided herein is a method of inhibiting IL-15-induced migration of T cells out of the skin (e.g., human skin), comprising contacting the skin with a therapeutically effective amount of an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9. In some embodiments, the T cells are CD8+ T cells. In some embodiments, the T cells are CD4+ T cells. In some embodiments, the skin is the skin of a subject having a disease or disorder associated with overexpression of CD122 or expression of CD122 in cells that do not normally express CD122.

[0109] The anti-CD122 antibodies or antigen-binding portions thereof described herein can be used in methods of treatment of the human or animal body, including prophylactic or preventative treatments (e.g., treatments to reduce the risk of, delay the onset of, or reduce the severity of a condition in a subject before the condition begins in the subject). Methods of treatment can include administering an anti-CD122 antibody or antigen-binding portion to a subject in need thereof. Provided herein are methods of treating or preventing a disease in a subject, comprising administering to the subject a therapeutically effective amount of an antibody, antigen-binding portion, immunoconjugate, or pharmaceutical composition disclosed herein.

[0110] Provided herein is a method for treating or preventing a disease in a subject, comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody, or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15. Provided herein is a method for treating or preventing a disease in a subject, comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody, or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9.

[0111] Provided herein is a method of alleviating, treating, or reducing the severity of a symptom of a disease in a subject, comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody, or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15. Provided herein is a method for alleviating, treating, or reducing the severity of a symptom of a disease in a subject, comprising administering to the subject a therapeutically effective amount of an anti-CD122 antibody, or antigen-binding portion thereof, wherein the antibody or antigen-binding portion comprises a VH region and a VL region, and (a) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:17, or (b) the amino acid sequence of the VH region comprises or consists of SEQ ID NO:1 and the amino acid sequence of the VL region comprises or consists of SEQ ID NO:9.

[0112] In some embodiments, the disease or disorder is associated with overexpression of CD122 or expression of CD122 in cells that do not normally express CD122. In some embodiments, the disease or disorder is mediated by CD122.

[0113] In some embodiments, the disease is inflammatory disease or autoimmune disease.In some embodiments, the disease is vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis, or rheumatoid arthritis.

[0114] In some embodiments, the VH region, the VL region, or both the VH and VL regions of an anti-CD122 antibody or antigen-binding portion used in the methods provided herein comprise one or more amino acid sequences of human framework regions.

[0115] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a pharmaceutical agent, e.g., an anti-CD122 antibody or antigen-binding portion thereof, sufficient to reduce or alleviate the severity and / or duration of a disease, e.g., vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis, or rheumatoid arthritis, or one or more symptoms thereof, prevent the progression of a disease, cause regression of a disease, prevent the recurrence, onset, onset, or progression of one or more symptoms associated with a disease, detect a disease, or enhance or improve the prophylactic or therapeutic effect(s) of another related therapy (e.g., a prophylactic or therapeutic agent) for a CD122-mediated disease.

[0116] The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of what is being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the composition, the method of administration, the administration schedule, and other factors known to the physician. Prescription of treatment, such as determining dosage, is within the responsibility of the general practitioner or other physician and may depend on the severity of the symptoms and / or progression of the disease being treated. Appropriate dosages of antibody molecules are well known in the art (Ledermann JA et al., 1991, Int. J. Cancer 47:659-664; Bagshawe KD et al., 1991, Antibody, Immunoconjugates and Radiopharmaceuticals 4:915-922). Specific dosages that may be indicated herein or in the Physician's Desk Reference (2003) as appropriate for the type of drug being administered may be used. A therapeutically effective amount, or suitable dosage, of an antibody molecule can be determined by comparing in vitro activity and in vivo activity in animal models. Methods for extrapolating effective dosages in mice and other test animals to humans are known. The exact dosage will depend on a number of factors, including whether the antibody is prophylactic or therapeutic, the size and location of the area to be treated, the precise nature of the antibody (e.g., whole antibody, fragment), and the nature of any detectable label or other molecule attached to the antibody.

[0117] Typical antibody doses will range from 100 μg to 1 g for systemic administration and 1 μg to 1 mg for intradermal injection. An initial larger dose may be administered, followed by one or more smaller doses. In some embodiments, the antibody is a whole antibody, e.g., IgG1 or IgG4 isotype. This is a single treatment dose for adult subjects, and can be adjusted accordingly for children and infants, as well as other antibody formats proportional to molecular weight. Treatments can be repeated at daily, twice-weekly, weekly, or monthly intervals, at the discretion of the physician. A subject's treatment schedule can depend on the pharmacokinetic and pharmacodynamic properties of the antibody composition, the route of administration, and the nature of the condition being treated.

[0118] Treatment can be cyclical, with the period between administrations being about 2 weeks or more, e.g., about 3 weeks or more, about 4 weeks or more, about once a month or more, about 5 weeks or more, or about 6 weeks or more. For example, treatment can be every 2-4 weeks, or every 4-8 weeks. Treatment can be pre-operative and / or post-operative, and / or can be administered or applied directly to the anatomical site of the surgical or invasive procedure. Suitable formulations and routes of administration are described above.

[0119] In some embodiments, the anti-CD122 antibodies and antigen-binding portions described herein can be administered as a subcutaneous injection, for example, using an auto-injector for long-term prophylaxis / treatment.

[0120] In some embodiments, the therapeutic effect of an anti-CD122 antibody or antigen-binding portion thereof may last for several half-lives, depending on the dose. For example, the therapeutic effect of a single dose of an anti-CD122 antibody or antigen-binding portion thereof may last for 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, or 6 months or more in a subject.

[0121] In some embodiments, a subject can be treated with an anti-CD122 antibody or anti-CD122 antigen binding portion, immunoconjugate or pharmaceutical composition described herein and an additional therapeutic agent or therapy used to treat a CD122-mediated disease or disorder, or a symptom or complication of a CD122-mediated disease or disorder. The anti-CD122 antibody or anti-CD122 antigen binding portion and the additional therapeutic agent or therapy can be administered simultaneously or sequentially.

[0122] In some embodiments, the subject is a human, a non-human primate, a pig, a horse, a cow, a dog, a cat, a guinea pig, a mouse, or a rat. In some embodiments, the subject is an adult human. In some embodiments, the subject is a pediatric human.

[0123] Further provided herein is an anti-CD122 antibody or anti-CD122 antigen-binding portion, immunoconjugate or pharmaceutical composition described herein for use in the treatment of a disease or disorder.

[0124] Provided herein is an anti-CD122 antibody or anti-CD122 antigen-binding portion, immunoconjugate or pharmaceutical composition described herein for use as a medicament.

[0125] definition Unless otherwise stated, the terms used herein have the definitions commonly used in the art. Some terms are defined below, and additional definitions can be found in the remainder of the detailed description.

[0126] The term "a" or "an" can mean one or more of the element, i.e., a plural referent. Thus, the terms "a," "an," "one or more," and "at least one" are used interchangeably herein. In addition, reference to an "element" by the indefinite article "a" or "an" does not exclude the possibility that more than one of the agents is present, unless the context clearly requires that one, and only one, of the agents is present.

[0127] Throughout this specification, the term "about" is used to indicate that a value includes the inherent variation of error for the device or method used to determine the value, or the variation that exists between samples measured. Unless otherwise specified or clear from the context, the term "about" means within 10% above or below the reported numerical value (except when such number is greater than 100% or less than 0% of the possible values). When used in conjunction with a range of values ​​or a series of values, the term "about" applies to both endpoints of the range of values ​​recited in the series, or each of them, unless otherwise indicated. As used in this application, the terms "about" and "approximately" are used as equivalents.

[0128] As used herein, the term "sequence identity" refers to the degree to which two optimally aligned polynucleotide or polypeptide sequences are invariant over an alignment window of residues, e.g., nucleotides or amino acids. The "fraction of identity" in reference to an aligned segment of a test sequence and a reference sequence is the number of identical residues shared by the two aligned sequences divided by the total number of residues in the reference sequence segment, i.e., the entire reference sequence or a more finely defined portion of the reference sequence. The "percent identity" is the fraction of identity multiplied by 100. Percent identity can be calculated using the alignment program Clustal Omega, available at ebi.ac.uk / Tools / msa / clustalo, with default parameters. See Sievers et al., "Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega" (2011 October 11) Molecular systems biology 7:539. For the purposes of calculating identity for sequences, extensions such as tags are not included.

[0129] As used herein, the term "HCDR" refers to heavy chain complementarity determining region. As used herein, the term "LCDR" refers to light chain complementarity determining region.

[0130] As used herein, the term "conservative substitution" refers to the replacement of an amino acid with another amino acid that does not significantly and adversely change its functional activity. A preferred example of a "conservative substitution" is the replacement of an amino acid with another amino acid having a value of > 0 in the following BLOSUM62 substitution matrix (see Henikoff & Henikoff, 1992, PNAS 89:10915-10919). TIFF2024513921000002.tif110164

[0131] By "antibody drug conjugate" and "immunoconjugate" is meant an antibody molecule or antigen-binding portion thereof, including antibody derivatives that bind to CD122 and are conjugated to a cytotoxic, cytostatic, and / or therapeutic agent.

[0132] The term "isolated molecule" (wherein the molecule is, for example, a polypeptide, polynucleotide, or antibody) refers to a molecule that, depending on its origin or source, is (1) not associated with naturally associated components with which it is naturally associated, (2) substantially free of other molecules of the same species, (3) expressed by cells of a different species, or (4) not occurring in nature. Thus, a molecule that is chemically synthesized or expressed in a cellular system different from the cells from which it is naturally derived would be "isolated" from its naturally associated components. A molecule can be rendered substantially free of naturally associated components by isolation using purification techniques well known in the art. The purity or homogeneity of a molecule can be analyzed by a number of means well known in the art. For example, the purity of a polypeptide sample can be assayed using polyacrylamide gel electrophoresis and staining the gel using techniques well known in the art to visualize the polypeptide. For certain purposes, higher resolution can be achieved using HPLC or other means well known in the art for purification.

[0133] The term "epitope" refers to a portion of a molecule capable of being recognized and bound by an antibody molecule or its antigen-binding portion at one or more of the antigen-binding regions of the antibody molecule. An epitope can be composed of a defined region of primary, secondary, or tertiary protein structure, and includes a combination of secondary structural units or structural domains of a target recognized by the antigen-binding region of an antibody or its antigen-binding portion. An epitope can also consist of a defined grouping of chemically active surface molecules, such as amino acids or sugar side chains, and can have specific three-dimensional structural characteristics, as well as specific charge characteristics. As used herein, the term "antigenic epitope" is defined as a portion of a polypeptide to which an antibody molecule can specifically bind, as defined by any method known in the art, for example, by conventional immunoassays, antibody competitive binding assays, or by x-ray crystallography or related structural measurements (e.g., nuclear magnetic resonance spectroscopy).

[0134] The term "potency" is a measure of biological activity and is expressed as IC 50 , E.C. 50 or as the effective concentration of an antibody or antibody-drug conjugate against the antigen CD122 that inhibits 50% of the activity as measured in the CD122 activity assay described herein.

[0135] As used herein with respect to the biological activity of the antibodies disclosed herein, the terms "inhibit" or "neutralize" refer to the ability of the antibody to substantially antagonize, inhibit, prevent, limit, slow, hinder, eliminate, stop, reduce, or reverse the progression or severity of what is being inhibited, including, but not limited to, the biological activity or binding interaction of the antibody molecule to CD122.

[0136] Any concentration range, percentage range, ratio range, or integer range herein should be understood to include any integer value within the stated range, and, where appropriate, fractions thereof (e.g., tenths and hundredths of integers), unless otherwise stated. The use of alternatives (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the terms "include" and "comprise" are used synonymously.

[0137] The headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0138] All documents or portions of documents cited herein, including but not limited to patents, patent applications, articles, books and papers, are expressly incorporated herein by reference in their entirety for all purposes. In the event that one or more of the incorporated documents or portions of documents defines a term in conflict with the definition of the term in this application, the definition set forth in this application shall prevail. However, the mention of any references, articles, publications, patents, patent publications, and patent applications cited herein is not, and should not be taken as, any form of acknowledgment or suggestion that they constitute valid prior art or form part of the general knowledge in any country in the world.

[0139] Any of the aspects and embodiments described in this specification may be combined with any other aspect or embodiment disclosed in the summary, drawings, and / or detailed description, including the specific non-limiting examples / embodiments of the disclosure below.

[0140] Numbered embodiments Without limiting the scope of the appended claims, the present disclosure describes the following numbered embodiments of the present disclosure.

[0141] 1. An anti-CD122 antibody, or antigen-binding portion thereof, comprising a heavy chain variable (VH) region and a light chain variable (VL) region; (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15, or (b) The antibody or antigen-binding portion thereof, wherein the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO: 3, an HCDR2 comprising SEQ ID NO: 5, and an HCDR3 comprising SEQ ID NO: 7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO: 11, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO: 15.

[0142] 2. (a) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:17, or (b) The antibody or antigen-binding portion of embodiment 1, wherein the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:9.

[0143] 3. The antibody or antigen-binding portion of embodiment 1 or 2, wherein said antibody or antigen-binding portion is humanized or chimeric.

[0144] 4. The antibody or antigen-binding portion of any one of embodiments 1 to 3, wherein the VH region, the VL region, or both the VH and VL regions comprise the amino acid sequence of one or more human framework regions.

[0145] 5. The antibody or antigen-binding portion of any one of embodiments 1 to 4, wherein the VH region, the VL region, or both the VH and VL regions comprise a human variable region framework scaffold amino acid sequence into which CDR amino acid sequences are inserted.

[0146] 6. The antibody or antigen-binding portion of any one of embodiments 1 and 3-5, wherein the VH region comprises an IGHV3-23 human germline scaffold amino acid sequence into which the HCDR1, HCDR2, and HCDR3 amino acid sequences are inserted.

[0147] 7. The antibody or antigen-binding portion of any one of embodiments 1 and 3-6, wherein the VL region comprises an IGKV1-33 human germline scaffold amino acid sequence into which the LCDR1, LCDR2, and LCDR3 amino acid sequences are inserted.

[0148] 8. The antibody or antigen-binding portion of any one of embodiments 1 to 7, wherein the antibody comprises an immunoglobulin constant region.

[0149] 9. The antibody or antigen-binding portion of embodiment 8, wherein the immunoglobulin constant region is IgG, IgE, IgM, IgD, IgA, or IgY.

[0150] 10. The antibody or antigen-binding portion of embodiment 9, wherein the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2.

[0151] 11. The antibody or antigen-binding portion of embodiment 8, wherein the immunoglobulin constant region is immunologically inert.

[0152] 12. The antibody or antigen-binding portion of embodiment 8, wherein said immunoglobulin constant region is a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG1 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A, or a wild-type human IgG2 constant region, wherein numbering is according to the EU index in Kabat.

[0153] 13. The antibody or antigen-binding portion of embodiment 8, wherein the immunoglobulin constant region comprises any one of SEQ ID NOs: 32-38.

[0154] 14. The antibody or antigen-binding portion of any one of embodiments 1 to 13, wherein the antibody or antigen-binding portion is a Fab, Fab', F(ab')2, Fv, scFv, maxibody, minibody, diabody, triabody, tetrabody, or bis-scFv.

[0155] 15. The antibody or antigen-binding portion of any one of embodiments 1 to 14, wherein the antibody is monoclonal.

[0156] 16. The antibody or antigen-binding portion of any one of embodiments 1 to 15, wherein the antibody is a tetrameric antibody, a tetravalent antibody, or a multispecific antibody.

[0157] 17. The antibody or antigen-binding portion of any one of embodiments 1 to 16, wherein the antibody is a bispecific antibody that specifically binds to a first antigen and a second antigen, wherein the first antigen is CD122 and the second antigen is not CD122.

[0158] 18. An immunoconjugate comprising the antibody or antigen-binding portion according to any one of embodiments 1 to 17, conjugated to a therapeutic agent.

[0159] 19. The immunoconjugate of embodiment 18, wherein the therapeutic agent is a cytotoxin, a radioisotope, a chemotherapeutic agent, an immunomodulatory agent, a cytostatic enzyme, a cytolytic enzyme, a therapeutic nucleic acid, an anti-angiogenic agent, an anti-proliferative agent, or a pro-apoptotic agent.

[0160] 20. A pharmaceutical composition comprising an antibody or antigen-binding portion according to any one of embodiments 1 to 17, or an immunoconjugate according to embodiment 18 or 19, and a pharma- ceutically acceptable carrier, diluent or excipient.

[0161] 21. The antibody or antibody binding portion according to any one of embodiments 1 to 17 (a) the amino acid sequence of the VH region; (b) the amino acid sequence of the VL region; or (c) both the amino acid sequences of the VH and VL regions; A nucleic acid molecule encoding

[0162] 22. An expression vector comprising the nucleic acid molecule of embodiment 21.

[0163] 23. A recombinant host cell comprising a nucleic acid molecule according to embodiment 21, or an expression vector according to embodiment 22.

[0164] 24. A method for producing an anti-CD122 antibody or antigen-binding portion thereof, comprising culturing a recombinant host cell comprising the expression vector of embodiment 22 under conditions in which the nucleic acid molecule is expressed, thereby producing the antibody or antigen-binding portion thereof, and isolating the antibody or antigen-binding portion thereof from the host cell or culture medium.

[0165] 25. A method for suppressing an immune response in a subject, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion of any one of embodiments 1 to 17, an immunoconjugate of embodiment 18 or 19, or a pharmaceutical composition of embodiment 20.

[0166] 26. The method of embodiment 25, wherein the immune response is mediated by CD122.

[0167] 27. A method for treating or preventing a disease in a subject, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion of any one of embodiments 1 to 17, an immunoconjugate of embodiment 18 or 19, or a pharmaceutical composition of embodiment 20.

[0168] 28. The method of embodiment 27, wherein the disease is an inflammatory disease or an autoimmune disease.

[0169] 29. The method of embodiment 27, wherein the disease is vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis, or rheumatoid arthritis.

[0170] 30. A method for inhibiting IL-15-induced T cell migration from the skin, comprising contacting the skin with a therapeutically effective amount of an antibody or antigen-binding portion of any one of embodiments 1 to 17, an immunoconjugate of embodiment 18 or 19, or a pharmaceutical composition of embodiment 20.

[0171] 31. An antibody or antigen-binding portion according to any one of embodiments 1 to 17, an immunoconjugate according to embodiment 18 or 19, or a pharmaceutical composition according to embodiment 20, for use as a medicament.

[0172] The present disclosure will be further clarified by the following examples, which are intended to be merely illustrative of the present disclosure and are not intended to limit the present disclosure in any manner. EXAMPLES

[0173] Example 1. Generation of an optimized anti-CD122 therapeutic antibody Introduction In this example, we successfully generate a panel of antagonistic, optimized anti-CD122 antibodies that are well expressed, biophysically stable, highly soluble, and have maximal source identity to the preferred human germline.

[0174] Materials and Methods Testing antibody v-domain specificity: Human receptor array analysis Human cell membrane receptor proteome arrays were performed at Retrogenix Ltd. Primary screening: 5 μg / mL of IgG1-MIKβ1 (humanized, also called VillMab-1) antibody was screened for binding to fixed HEK293 cells / slide (14 slide sets, n=2 slides per slide set) expressing 4975 individual human plasma membrane proteins. All transfection efficiencies exceeded a minimum threshold. Antibody binding was detected using AF647 fluorescent secondary anti-human IgG1 antibody. Primary hits (overlapping spots) were identified by analyzing fluorescence (AF647 and ZsGreen1) in ImageQuant. Vectors encoding all hits were sequenced to confirm their exact identity.

[0175] Confirmation / specificity screen: Vectors encoding all hits, plus control vectors encoding MS4A1 (CD20) and EGFR, were spotted in duplicate on new slides and used to reverse transfect human HEK293 cells as described above. All transfection efficiencies exceeded the minimum threshold. Identical fixed slides were treated (n=2 slides per treatment) with 5 μg / mL of each test antibody, 5 μg / mL of negative control antibody, 1 μg / mL of rituximab biosimilar (positive control), isotype IgG1 (Ab00102 human IgG1 anti-fluorescein), or no test molecule (secondary only, negative control). Slides were analyzed as described above.

[0176] Flow cytometry confirmation screen: Human HEK293 cells were transfected with expression vectors encoding ZsGreen1 alone or ZsGreen1 and CD122, BCAM. Live transfectants were incubated with 1 and 5 mg / mL of each of the test and isotype control antibodies. Cells were washed and incubated with the same AF647 anti-human IgG Fc detection antibody used in the cell microarray screen. Cells were washed again and analyzed by flow cytometry using an Accuri flow cytometer (BD). Dead cells were excluded using 7AAD live / dead dye, and ZsGreen1 positive cells (i.e., transfected cells) were selected for analysis.

[0177] CD122 library generation and selection CD122 Fab repertoires were constructed by mass oligo synthesis and PCR. The amplified Fab repertoires were then cloned into phagemid vectors by restriction ligation, transformed into E. Coli TG-1 cells, and phage repertoires were essentially rescued as previously detailed (Finlay et al., 2011, Methods Mol Biol 681:383-401). Phage selection was performed by coating streptavidin magnetic microbeads with biotinylated CD122 target protein (either human or cynomolgus monkey) and washing the beads three times with PBS and resuspending them in PBS (pH 7.4 + 5% skim milk protein). The beads were coated with 100 nM of target protein in the first round of selection, followed by three successive rounds with decreasing concentrations of antigen. In each round, phages were reinfected into TG1 cells after elution with trypsin.

[0178] Fab and IgG expression and purification Mammalian codon-optimized synthetic genes encoding the heavy and light chain variable domains of the lead panel anti-CD122 antibody, plus MIKβ1 variants, were cloned into mammalian expression vectors containing effector function null human IgG1 ("IgG1-3M": human IgG1 containing L234A, L235A, G237A mutations in the lower hinge that abolish normal immunoglobulin ADCC, ADCP, and CDC functions), and human Cκ domain, respectively. Co-transfection of heavy and light chain containing vectors in the mammalian expression system was followed by Protein A-based IgG purification, quantification, and QC by denaturing and non-denaturing SDS-PAGE.

[0179] Direct binding ELISA for Fab and IgG Binding and cross-reactivity of the lead panel to recombinant proteins was first assessed by binding ELISA. Human CD122 human Fc tag recombinant proteins and cynomolgus and / or rhesus CD122 human Fc tag recombinant proteins were coated onto the surface of MaxiSorp™ flat-bottom 96-well plates at 1 μg / mL. Purified Fab or IgG samples were titrated in two-fold serial dilutions starting from 500 nM to 0.98 nM and allowed to bind to the coated antigen. Fab was detected with mouse anti-c-myc antibody followed by donkey anti-mouse IgG conjugated to horseradish peroxidase. IgG was detected with mouse anti-human IgG conjugated to horseradish peroxidase. Binding signals were visualized with 3,3'5,5'-tetramethylbenzidine substrate solution (TMB) and absorbance was measured at 450 nm.

[0180] BIACORE™ Analysis of Fab Affinity to Human and Rhesus CD122 The affinity (KD) of purified IgG was measured by SPR with antigen in solution on a BIACORE® 3000 (GE). Mouse anti-human antibody (CH1 specific) was immobilized to a CM5 sensor chip at 2000 RU in acetate buffer at pH 4.5 using amine coupling according to the Wizard instructions for two channels. One channel was used for background signal correction. Standard running buffer HBS-EP (pH 7.4) was used. Regeneration was performed with a single injection of 10 μL of 10 mM glycine at pH 1.5 at 20 μL / min. IgG samples were injected at 50 nM at 30 μL / min for 2 min followed by a 60 s dissociation rate. Monomeric antigen (human CD122 His-tag, or cynomolgus and / or rhesus CD122 His-tag) was injected at 30 μL / min for 2 min in 2-fold serial dilutions from 100 nM down to 3.1 nM, followed by a dissociation rate of 300 s. The resulting sensorgrams were analyzed using BIACORE® 3000 evaluation software. KD was calculated by simultaneously fitting the association and dissociation steps to a 1:1 Langmuir binding model.

[0181] IgG flow cytometry Purified IgG was tested by FACs for binding to human and rhesus CD122 expressed in CHO-K1 stable cell lines and CHO-K1 wild type cells. IgG samples were titrated in 3-fold serial dilutions starting from 500 nM to 0.98 nM. IgG binding was detected using mouse anti-human IgG conjugated with FITC. Results were analyzed by examining the mean fluorescence intensity (MFI) of 10,000 cells per sample on the BL-1 channel detector of a flow cytometer (Attune™ NxT Acoustic Focusing Cytometer, Invitrogen / ThermoFisher Scientific).

[0182] M07e cell line assay M07e cells were obtained from the DSMZ-German collection of microorganisms and cell culture media and maintained in RPMI supplemented with 10% FBS, 10 ng / mL GM-CSF (Peprotech), and L-glutamine (Corning) according to guidelines provided by the distributor. On day 1, cells were washed in RPMI and cultured at 2.5 × 10 5 The cells were resuspended at a density of 5.0 × 10 cells / mL in a final volume of 200 μL. 4 Cells were cultured in wells of 96-well flat-bottom plates in the presence of 50 ng / mL recombinant human IL15 (rhIL15) (R&D), or rhIL15 plus antibody for 72 h at 37°C. After 72 h, cells were incubated with 20 μL of WST-1 cell proliferation reagent (Miltenyi) for 3 h at 37°C. Cell proliferation was quantified using a scanning multiwell photometer, and absorbance measured at 450 nM was corrected for the number of viable cells.

[0183] Human NK cell line assay Human peripheral blood mononuclear cells (PBMCs) were isolated from donor whole blood using density gradient centrifugation over Ficoll Histopaque, and NK cells were enriched from isolated PBMCs using a Miltenyi Biotec (Bergisch Gladbach, Germany) human NK cell isolation kit according to the manufacturer's instructions. NK cells were stained with the CellTrace™ CFSE cell proliferation kit according to the manufacturer's instructions and resuspended in RPMI supplemented with 10% FBS and penicillin-streptomycin (Gibco). A total of 10 5NK cells were cultured in wells of a 96-well round-bottom plate in the presence of 20 ng / mL recombinant human IL15 (rhIL15) (R&D), or rhIL15 plus antibody for 120 h at 37° C. After 120 h, NK cells were washed and stained with anti-human CD3 (UCHT1), CD56 (5.1H11), and CD16 (3G8) (1:20 dilution, Biolegend) and analyzed for CFSE dilution on a BD LSR II flow cytometer (BD Biosciences) and FlowJo (Tree Star Inc.).

[0184] NSG-IL15 mouse model Humanized mice were generated by transplanting human hematopoietic stem cells (HSCs) into NOD-squid gamma mice expressing human IL15 (NSG-Tg). Six to eight week old NSG-Tg mice were first irradiated with 200 cGy and then transplanted with 1000 ng / mL of umbilical cord blood-derived IL15. 5 CD34+ HSCs were injected into the mice. HSC-engrafted NSG-Tg mice were screened at 12 and 16 weeks to measure baseline engraftment in blood. NSG-Tg mice with more than 20% human CD45+ cells with more than 2% CD56+ were selected for antibody treatment. Mice were treated with intraperitoneal (ip) injections twice a week (Monday / Thursday schedule) for 3 weeks. The amount of human immune cells in the blood was quantified using flow cytometry at 1 and 3 weeks after the start of treatment, and then at 1, 3, and 5 weeks after treatment. At 5 weeks after treatment, mice were euthanized and the amount of human immune cells in the spleen and blood was measured by flow cytometry. Cells from all tissues were stained with anti-human CD45, CD3, CD4, CD8, CD7, CD56, CD16, Mik-b2, and Mik-b3 (1:20 dilution, Biolegend) and analyzed on a BD LSR II flow cytometer (BD Biosciences) and FlowJo (Tree Star Inc.).

[0185] Results and Discussion Pharmacological modeling: Assessing the feasibility of IL15Rβ antagonist antibodies for treating diseases in the skin In silico modeling was performed to define pharmacological parameters and properties that may confer therapeutic success for anti-CD122 IgG1 antibodies designed to maximize efficacy in specific tissues such as the skin. These analyses were based on established values ​​for CD122 target biology (Table 1) and known potential drug properties and dosing parameters for bivalent IgG antibodies (Table 2).

[0186] The parameters outlined above were then analyzed to sample the impact on drug potency and distribution as outlined in Figure 1. The bottom-up estimation approach was validated using reported PK data from t-cell leukemia patients previously treated with the humanized anti-CD122 IgG1 antibody "MIKβ1." These analyses provided a set of insights regarding IV and / or SC administration and antibody CD122 binding affinity in weight-average human subjects.

[0187] At an apparent drug affinity of 1.10 nM KD, sustained inhibition (>90%) of IL15Rβ in the skin was predicted to be achievable with 700 mg IV Q4W, or 100 mg SC Q1W (Table 3, Table 4).

[0188] 2. Functional affinity above 10 nM KD allows for lower dosing requirements for IV and SC, with KD values ​​closer to 1 nM allowing 99% target occupancy in the skin at the highest doses. This pattern is qualitatively similar for the IV and SC routes of administration (Table 3, Table 4).

[0189] 3. At an apparent drug binding affinity of 10 nM, 100 mg of SC Q1W is sufficient to maintain 90% receptor occupancy in the skin, but 100 mg of SC Q1W is not sufficient to maintain >95% receptor occupancy. Higher affinity would be required to achieve >95% receptor occupancy with the 100 mg SC Q1W dosing schedule (Table 4).

[0190] 4. Eliminating systemic target sinking has minimal impact on the dose required to achieve high (>90%) target occupancy in the skin. Simulations of IV Q4W dosing in the presence and absence of systemic CD122 at known concentrations suggest that the total dose must overcome the systemic target burden to have optimal activity in the skin and drug distribution effects (Table 5).

[0191] Collectively, these analyses suggest that an optimal anti-CD122 antagonistic therapeutic antibody would have a functional affinity for CD122 above 10 nM, allowing dosing at 700 mg IV Q4W or 100 mg SC Q1W. Increased affinity would allow greater target coverage at lower doses.

[0192] Antibody binding specificity analysis In early clinical trials, the humanized anti-CD122 IgG1 antibody "MIKβ1" has been reported to show evidence of accelerated clearance. Accelerated clearance is a risk factor that may prevent the achievement of the ideal drug properties outlined above due to the possibility that target-mediated drug distribution (TMDD) effects may negatively impact efficacy. We hypothesized that MIKβ1 may not be able to bind exclusively to CD122, but may bind to unidentified and unpredictable human proteins. To investigate this possibility, we screened for off-target binding specificity in humanized IgG1-MIKβ1 (VillMab-1) using an in vitro technique (Retrogenix, Ltd.) based on the use of high-density arrays of cells expressing over 5500 unique human membrane receptors and membrane-tethered secreted proteins. Our receptor array binding screen identified that VillMab-1 showed strong binding to membrane-expressed CD122, but also had potential off-target binding specificity to BCAM (also known as AU, CD239, LU, MSK19, basal adhesion molecule (Lutheran blood group)), a widely expressed membrane adhesion protein that may cause reduced PK and enhance the "sinking" effect of therapeutic doses of anti-CD122 antibodies.

[0193] To confirm this off-target binding event, plasmids encoding BCAM and control proteins were subjected to DNA sequencing. These analyses confirmed that the encoded proteins were indeed of the correct sequence. Plasmid samples for the control and potential target receptors were then rearranged onto new chips for replicate analyses in duplicate. By scanning the chips for ZS green, which is simultaneously encoded in all expression plasmids as an internal control marker, it was confirmed that efficient expression was induced from all rearranged plasmids. This analysis showed clearly detectable ZS expression at all positions where the plasmids were spotted (Figure 2A). Furthermore, identically spotted slides were then used to reprobe cells transfected with VillMab-1 (Figure 2B), rituximab (IgG1 positive control, Figure 2C), and a chip where no primary antibody probe was applied (Figure 2D). These analyses showed that VillMab-1 again showed measurable binding above background (in both chips) in BCAM-transfected cells (Figure 2B). Rituximab showed binding to CD20 as expected, with no observable binding to any other proteins (Figure 2C). In chips not probed with primary antibodies (Figure 2D), only the expected control proteins showed any signal. This clear state of the control chips confirmed that the VillMab-1 binding signals at CD122 and BCAM were specific. To further confirm these off-target binding results, the sequence-verified plasmids were again transfected into HEK-293 cells and binding was examined by flow cytometry (Figure 3). In this experiment, VillMab-1 showed clear binding to both CD122 and BCAM transfected cells, but no background binding to ZS-transfected cells ("ZS only", Figure 3A).Control experiments using the same cells but without primary antibody (Figure 3B) showed that the BCAM signal was antibody associated, and staining with rituximab IgG1 anti-CD20 (Figure 3C) showed binding signal only in CD20-transfected cells, demonstrating that the signal at BCAM is specifically mediated by the binding domain of VillMab-1.

[0194] Because off-target binding has been shown to have a negative impact on the PK, biodistribution, and toxicity profile of IgG, therapeutic antibodies should ideally have excellent specificity for the desired target. To address this issue in VillMab-1, experimental modulation of the VillMab-1 binding interface was performed as follows.

[0195] Mutagenesis and paratope modulation of VillMab-1 To bias our recombination efforts towards a final lead therapeutic IgG compound with optimal drug-like properties, we chose to explore mutation-derived variants of the VillMab-1 antibody. Sequence analysis of the VillMab-1 v-domain showed that the original humanization process used scaffolds related to the human germline frameworks IGHV3-23 and IGKV1-33, known to have good solubility and drug development quality and frequently used in the expressed human antibody repertoire (Table 6). Despite this use of well-known scaffolds, both variable domain frameworks contained a significant number of deviations from the germline sequence. In addition, the CDR sequences also contained many residues that differed from the human germline (Table 6).

[0196] The VillMab-1 v-domain sequence was combined with a Fab phage display format and separate mutagenesis library cassettes were generated for the VH and VL domains by oligo synthesis and assembly. Each mutagenesis cassette encoded the VillMab-1 residue, the human germline residue, or the homologous amino acid at each position underlined in Table 6. Separate Fab libraries were generated by combining the mutagenesis cassette for VL with VillMab-1 VH or VillMab-1 VL with the mutagenesis cassette for VH. Each final Fab library was ligated into a phage display vector and transformed into E. coli by electroporation for 10 min. 7 More than 100 independent clones were generated. The quality of library construction was verified by sequencing 96 clones per library. The sequencing data showed that the mutated positions efficiently sampled the designed diversity. The library was rescued with helper phage M13 and selections were performed on biotinylated human and cynomolgus and / or rhesus CD122-Fc proteins in multiple separate branches. After one round of selection, a third combinatorial library was generated using combinations of preselected mutated VH and VL to simultaneously sample the selected variability in both V domains.

[0197] Post-selection plasma periprep screening and DNA sequencing revealed the presence of 64 unique human and rhesus CD122 binding Fab clones that showed strong binding to human and rhesus CD122 in ELISA and more than 50% inhibition of VillMab-1 binding to human and rhesus CD122 in Alphascreen assays. From these unique library-derived leads, the top 15 clones were identified based on assay signal strength, mutability relative to human germline, and absence of major developmental defects / chemical degradation motifs (Table 7). This analysis also identified a set of unique sequences in each CDR (Table 8). These unique CDR sequence profiles were used to design an additional 15 clones (MAB01-MAB15) with potential CDR combinations not found among the top 15 library-derived clones as outlined in (Table 9). A total of 30 of these unique clones were expressed in human IgG1 format in CHO transient cultures, purified by Protein A, and confirmed to be >95% monomeric by size exclusion chromatography.

[0198] Specificity and potency characteristics of lead IgG Next, the purified IgGs described above were tested for competition for the VillMab-1 binding epitope on human CD122-Fc in Alphascreen format. This analysis showed that 24 of the 30 clones exhibited a concentration-dependent reduction in VillMab-1 binding to CD122, proving that they retained the functional epitope of the parent antibody (Figure 4). These 24 clones were then investigated in a polyreactivity assay to confirm that the final lead clones from the first recombination did not have DNA or insulin binding profiles strongly associated with short PK (Figure 5). This analysis showed that, although all 24 clones showed significantly lower signals than the positive control bococizumab, a subset produced particularly low signals that were equal to or improved over the negative control antibodies bevacizumab, ustekinumab, and pembrolizumab (Figure 5). These findings allowed six key leads to be prioritized for further analysis.

[0199] Concentration-dependent binding of six prioritized lead clones from the library to human and rhesus CD122 at the cell surface was analyzed by flow cytometry (Figure 6). Clones 06F11 (Figure 6A), 07C07 (Figure 6B), 07D06 (Figure 6C), 07E09 (Figure 6D), 07D07 (Figure 6E), and 06D12 (Figure 6F) each exhibited CD122-specific binding profiles with binding curves very similar to those observed for VillMab-1, while the isotype control IgG1 showed no binding to any cell type (Figure 6G).

[0200] Analysis of lead IgG in the CD122-IL-15 blocking assay In the M07e cell-based CD122 / IL-15 blocking reporter assay, clones 06F11 (Figure 7A), 07C07 (Figure 7B), 07D06 (Figure 7C), 07E09 (Figure 7D), 07D07 (Figure 7E), and 06D12 (Figure 7F) showed concentration-dependent antagonism of CD122. The IC50 for VillMAB-1 was 9.792 μg / mL. The IC50 for 6F11 and 7C07 was 14.8 μg / mL and 20.8 μg / mL, respectively, the lowest among the lead clones. The IC50 for 06D12, 07D06, 07D07, and 07E09 was 38.610 μg / mL, 27.820 μg / mL, 34.170 μg / mL, and 23.610 μg / mL, respectively. This analysis highlighted 06F11 and 07C07 as ideal candidates for further evaluation.

[0201] Analysis of lead Fabs in CD122-bound BIACORE® for 1:1 binding affinity To characterize the 1:1 affinity values ​​for clones 06F11 and 07C07, as well as a variant of 06F11 with a corrected mutation in FW1 (06F11-V), these and the positive control Villmab-1 clone were cloned, expressed and purified in human Fab format (i.e., monovalent and lacking both hinge and Fc regions). Binding of the fully purified Fab proteins to both human CD122 (Table 10) and rhesus CD122 (Table 11) was investigated. These analyses showed that clones FAB06F11-V, FAB06F11, and FAB07C07 showed overall moderately reduced KD values ​​for both human and rhesus CD122 compared to VillFab-1, and importantly, also showed increased on (ka) and off (kd) rates.

[0202] Analysis of lead IgG in the hIL-15 NSG mouse model Using a humanized hIL-15 NSG mouse model, we compared the ability of VillMAB-1, 7C07, and 6F11-v (all in an IgG1-3M effector null format) to inhibit engraftment of human NK and CD8 T cells supported by CD122 / IL15 signaling in vivo. After establishing complete engraftment with the cells, mice were treated with vehicle, low dose (1 mg / kg), or high dose (10 mg / kg) antibody.

[0203] Prior to antibody treatment, the numbers of human CD8+ T cells (Figure 8A) and NK (Figure 8B) cells in the blood were similar between the groups of mice. After one week of treatment, only mice treated with 10 mg / kg VillMAB-1 showed a statistically significant decrease in the number of CD8+ T cells compared to isotype-treated mice (Figure 8C). A decrease in CD8+ T cells was observed in all antibody-treated groups when compared to isotype-treated mice. The number of NK cells in the blood was similar in all groups (Figure 8D).

[0204] After 3 weeks of treatment, the number of CD8+ T cells was reduced in the blood of all antibody-treated groups compared to isotype-treated mice (Figure 8E). These changes were not statistically significant. The number of NK cells was reduced in all groups of antibody-treated mice, and this reduction was statistically significant in all groups except for mice treated with 1 mg / kg 07C07 (Figure 8F). High and low dose VillMAB-1 treatment reduced the number of NK cells more than 06F11-V or 07C07 treatment at either dose.

[0205] Importantly, these findings confirmed that chronic blockade of CD122-IL15 signaling with clones Villmab-1, 06F11-V, or 07C07 was capable of depleting both NK and CD8+ t cell populations in the absence of ADCC or ADCP effector function.

[0206] Analysis of antibody binding specificity of clones 06F11-V and 07C07 As clones 06F11-V and 07C07 have shown above to have a high level of humanization in both the framework regions and CDRs of the v-domain, as well as effective blocking of CD122 in vitro and in vivo, they were screened again for specificity analysis in the Retrogenix proteomics platform. Unexpectedly, this analysis showed that the BCAM binding observed for Villmab-1 was completely abolished and was not observed for either 06F11-V or 07C07 IgG. However, in addition, two new interactions were observed that were not observed for Villmab-1 (Figure 9). In flow cytometry analysis of binding to cells transfected with plasmids driving the expression of the target and control, in addition to CD122 binding, clone 06F11-V was found to bind both neudecin (neurotropin) and CILP2 (cartilage structural protein), while 07C07 was found to bind only CILP2 (Figures 9A, 9B). In contrast, "secondary antibody only" (Figure 9C) and rituximab primary antibody (Figure 9D) control experiments showed either no binding above background or binding only to CD20 transfected cells, respectively. These findings confirmed that the off-target binding observed for 06F11-V and 07C07 IgG was genuine and specific.

[0207] Optimization of clone 06F11-V Clone 06F11-V was selected for further optimization to maximize beneficial properties and minimize off-target binding. This optimization was performed by experimentally analyzing combinations of mutations back to mouse sequences in CDR1, and / or 2, and / or 3 of both the heavy and light chain sequences of 06F-11V. This process generated 18 new clones, each with one of six VH sequences combined with one of three VL sequences (Table 12). These 18 new variants, Villmab-1 and 06F11V, were cloned in human monovalent Fab fragment format, expressed in CHO cells, and purified to monomeric state by Protein A column followed by SEC.

[0208] Next, the binding affinity of the purified Fabs to human and rhesus CD122 was investigated by BIACORE® (Table 13). This analysis showed that a series of clones showed improved affinity to CD122 over 06F11-V and even over VillMab-1. From this cohort, clones MAB05, MAB06, MAB14, MAB15, MAB17, and MAB18 were prioritized for further analysis and were further expressed and purified in human IgG1-3M format for potency and specificity analysis. When the Fab versions of all prioritized clones were tested by BIACORE® binding on human neudecin and CILP2 proteins (Figure 10A, Figure 10B), only clone 06F11-V was found to show measurable binding to either protein, suggesting that the new lead clones eliminated these two off-target binding risks. In ELISA analysis of IgG against all clones in human BCAM protein, all clones retained off-target binding except MAB05 and MAB06, and showed perfect specificity for CD122 (Figure 10C, Table 14).

[0209] To investigate how such closely related antibody sequences could have such radical differences in specificity profiles, we performed sequence alignments of both VH and VL domains of VillMab-1 to clones 05, 06, 14, 15, 17, and 18 (Figures 11A, 11B). Of note, the only changes between the unique VillMab-1 sequence to (fully CD122-specific) clones MAB05 and MAB06 were found to be three (highly homologous) mutations in and near CDR1 of the VH domain. This finding illustrates the promiscuous and unpredictable nature of antibody binding.

[0210] Confirmation of biological potency of clones MAB05, MAB06, MAB14, MAB15, MAB17, and MAB18 was performed in an IL-15 stimulated M07e assay (Table 14). This analysis showed that not only was the affinity of clones MAB05 and MAB06 improved over VillMab-1, but their potency in blocking IL-15 signaling was also improved by about 2-fold (Table 14). The final functionally relevant potency of clones MAB05 and MAB06 was then confirmed in an assay measuring the proliferation of human primary NK cells under IL-15 stimulation (Figures 12A, 12B). This assay reproduced the findings in the M07e assay, showing that MAB05 and MAB06 are indeed improved over VillMab-1.

[0211] IgG1-3M BIACORE® Analysis of Binding to Human and Mouse Fc Receptors To characterize the affinity of clone 06F11-V IgG1-3M and the positive control IgG, the binding of the fully purified protein to both human and mouse FcγR and FcRn was investigated (Table 17). These analyses showed that while all positive controls showed the expected strong interactions with both human and mouse receptors, clone 06F11-V IgG1-3M showed very low or no measurable affinity to either human or mouse FcγR and FcRn at pH 7.4. Importantly, however, 06F11-V IgG1-3M retained full affinity for human and mouse FcRn at pH 6.0. These findings confirmed what was observed in vivo in the NSG / IL-15 mouse model, that blocking CD122 signaling in the absence of IgG effector function is sufficient to deplete CD122+ cells.

[0212] Example 2. Function of anti-CD122 therapeutic antibodies in human cutaneous T cell crawl-out assays A human skin biopsy culture assay was used to investigate the ability of MAB05 and MAB06 to inhibit CD122 / IL-15 signaling in skin-resident T cells.

[0213] Human skin biopsies (4 mm diameter × 2 mm thickness) were collected from surgical specimens (subcutaneous adipose tissue excision) using a 4 mm Integra disposable biopsy punch (Integra). Skin biopsies were incubated in Antibiotic-Antimycotic (Gibco) diluted in PBS for 30 min at 4°C, followed by 3 rinses with PBS. Three skin biopsies were placed into one well of a 24-well plate (Corning) and briefly dried to promote adhesion of the biopsies to the well surface. Biopsies were then cultured in 2 mL of Iscove's modified medium (Sigma) containing 20% ​​heat-inactivated fetal bovine serum, penicillin and streptomycin (Corning), and 3.5 μL / L β-mercaptoethanol (Sigma) and incubated at 37°C for 21 days. Cultures were fed three times a week by aspirating 1 mL of medium from each well and replacing with 1 mL of fresh medium. For cultures treated with IL-15 or anti-CD122 antibodies, 20 ng / mL recombinant human IL15 (rhIL15) (R&D) and anti-CD122 antibodies (MAB05 or MAB06) were added from the initiation of culture until T cell harvest on day 21. After 21 days, culture medium was collected from wells and spun down at 330×g for 10 min in 5 mL polystyrene round-bottom tubes. Supernatants were aspirated and remaining T cells were washed and stained with anti-human CD3, CD4, and CD8 (1:20 dilution, Biolegend) and quantified using a BD LSR II flow cytometer (BD Biosciences) and FlowJo (Tree Star Inc.).

[0214] In this assay, MAB05 and MAB06 inhibit IL15-induced accumulation of CD8+ T cells migrating from skin biopsies. When biopsies are cultured with IL-15, more CD8+ T cells accumulate than when biopsies are cultured without IL-15: 11,101±6011 vs. 438.3±66.05 (mean±SD) (FIG. 13A). When biopsies are cultured with IL-15 and MAB05, fewer CD8+ T cells accumulate compared to cultures with IL-15 alone: ​​2096±1100 vs. 11,101±6011 (mean±SD) (FIG. 13A). Similarly, when biopsies are cultured with IL-15 and MAB06, fewer CD8+ T cells accumulate compared to cultures with IL-15 alone: ​​2436±501.6 vs. 11,101±6011 (mean±SD) (FIG. 13A).

[0215] MAB05 and MAB06 also inhibit IL15-induced accumulation of CD4+ T cells migrating from skin biopsies. When biopsies are cultured with IL-15, more CD4+ T cells accumulate than when biopsies are cultured without IL-15: 40,523±15,391 vs. 1261±473.6 (mean±SD) (FIG. 13B). When biopsies are cultured with IL-15 and MAB05, fewer CD4+ T cells accumulate compared to cultures with IL-15 alone: ​​3471±1627 vs. 40,523±15,391 (mean±SD) (FIG. 13B). Similarly, fewer CD4+ T cells accumulated when biopsies were cultured with IL-15 and MAB06 compared with cultures with IL-15 alone: ​​4308±2111 vs. 40,523±15,391 (mean±SD) (FIG. 13B).

[0216] In skin biopsy culture assays, MAB05 demonstrated concentration-dependent antagonism of IL-15-induced accumulation of skin-resident CD8+ T cells with an IC50 of 1.9 μg / mL (FIG. 14A). MAB06 demonstrated comparable concentration-dependent antagonism of CD8+ T cell accumulation with an IC50 of 1.8 μg / mL (FIG. 14A). Furthermore, MAB05 and MAB06 demonstrated comparable concentration-dependent antagonism of IL-15-induced CD4+ T cell accumulation with IC50s of 2.1 μg / mL and 1.8 μg / mL, respectively (FIG. 14B).

[0217] Example 3. Pharmacokinetic / pharmacodynamic (PK / PD) study of anti-CD122 therapeutic antibodies in cynomolgus monkeys Cynomolgus monkeys were administered a single intravenous injection of anti-CD122 antibodies (MAB05 or MAB06) at dose levels ranging from 1 to 20 mg / kg. Blood samples were collected before and at various time points ranging from 1 hour to 16 days after dosing. Pharmacokinetic parameters were measured following quantification of anti-CD122 antibody plasma concentrations by ELISA. Blood samples were analyzed for CD122 receptor occupancy on NK cells using flow cytometry, and total T cells, helper T cells, cytotoxic T cells, and NK cells were quantified.

[0218] Following single intravenous administration at dose levels ranging from 1 mg / kg to 20 mg / kg, MAB05 and MAB06 showed no clinical abnormalities and showed linear pharmacokinetics in cynomolgus monkeys. A single dose of 1 mg / kg of MAB05 and MAB06 was sufficient to maintain >90% CD122 receptor occupancy on NK cells throughout the sampling period (16 days post-dose). Administration of single doses of 1 mg / kg, 5 mg / kg, or 20 mg / kg of MAB05 and MAB06 induced a decrease in circulating NK cell numbers. Circulating NK cells reached a nadir approximately 7 days after dosing and showed a stable recovery after dosing of 1 mg / kg or 5 mg / kg throughout the remaining sampling period (16 days post-dose). Modulation of the number of circulating NK cells is considered a marker of functional activity and is important in demonstrating that anti-CD122 antibodies are effective in vivo (see Waldmann et al. (2020) J Exp Med 217:e20191062), and can therefore be advantageously used to define optimal dosing in patients. No effect was observed with helper T cells or cytotoxic T cells.

[0219] Example 4. Preformulation development and stability studies of anti-CD122 therapeutic antibodies A panel of stability studies was used to evaluate the manufacturability of anti-CD122 antibody. Ten formulations of anti-CD122 antibody in combinations of buffers and excipients were investigated for stability and aggregation potential at a concentration of 5 mg / mL. The antibody was investigated under conditions of low pH stress, heat stress, freeze-thaw conditions, and forced oxidation. The antibody was also investigated for self-association and viscosity at concentrations >100 mg / mL.

[0220] MAB05 and MAB06 showed high thermal stability, high resistance to low pH (pH 3.0), low aggregation potential, low oxidative and amidolytic sensitivity, and excellent freeze / thaw stability in a buffer solution (pH 6.0) with the composition of 25 mM L-histidine, 9% (w / v) sucrose, and 0.02% (w / v) polysorbate 80. MAB05 and MAB06 were solubilized in this same buffer solution at concentrations ranging from 90 to 120 mg / mL. The viscosity of MAB05 and MAB06 at a concentration of 115 mg / mL was low (approximately 4 to 6 centipoise). This indicates that it is possible to realize a clinical formulation for subcutaneous delivery of anti-CD122 antibodies. [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7-1]

Table 7-2

Table 8-1

Table 8-2

Table 8-3

Table 8-4

Table 9

Table 10

Table 11

Table 12-1

Table 12-2

Table 13

Table 14

Table 15-1

Table 15-2

Table 16

Table 17

Table 18

Table 19

Table 20

Claims

1. An anti-CD122 antibody, or antigen-binding portion thereof, comprising a heavy chain variable (VH) region and a light chain variable (VL) region; (a) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:18, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15; or (b) the amino acid sequence of the VH region comprises an HCDR1 comprising SEQ ID NO:3, an HCDR2 comprising SEQ ID NO:5, and an HCDR3 comprising SEQ ID NO:7, and the amino acid sequence of the VL region comprises an LCDR1 comprising SEQ ID NO:11, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:15; The antibody or antigen-binding portion thereof.

2. (a) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:17; or (b) the amino acid sequence of the VH region comprises SEQ ID NO:1 and the amino acid sequence of the VL region comprises SEQ ID NO:9; The antibody or antigen-binding portion of claim 1.

3. The antibody or antigen-binding portion of claim 1 , wherein the antibody or antigen-binding portion is humanized or chimeric.

4. 2. The antibody or antigen-binding portion of claim 1, wherein the VH region, the VL region, or both the VH and VL regions comprise one or more amino acid sequences of human framework regions.

5. 2. The antibody or antigen-binding portion of claim 1, wherein the VH region, the VL region, or both the VH and VL regions comprise a human variable region framework scaffold amino acid sequence into which CDR amino acid sequences are inserted.

6. 2. The antibody or antigen-binding portion of claim 1, wherein the VH region comprises an IGHV3-23 human germline scaffold amino acid sequence into which the HCDR1, HCDR2, and HCDR3 amino acid sequences are inserted.

7. 2. The antibody or antigen-binding portion of claim 1, wherein the VL region comprises an IGKV1-33 human germline scaffold amino acid sequence into which the LCDR1, LCDR2, and LCDR3 amino acid sequences are inserted.

8. The antibody or antigen-binding portion of claim 1 , wherein the antibody comprises an immunoglobulin constant region.

9. 9. The antibody or antigen-binding portion of claim 8, wherein the immunoglobulin constant region is IgG, IgE, IgM, IgD, IgA, or IgY.

10. 10. The antibody or antigen-binding portion of claim 9, wherein the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2.

11. The antibody or antigen-binding portion of claim 8, wherein the constant region of the immunoglobulin is immunologically inert.

12. 9. The antibody or antigen-binding portion of claim 8, wherein the immunoglobulin constant region is a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG1 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, and G237A, or a wild-type human IgG2 constant region, numbering according to the EU index in Kabat.

13. The antibody or antigen-binding portion of claim 8, wherein the immunoglobulin constant region comprises any one of SEQ ID NOs: 32 to 38.

14. The antibody or antigen-binding portion is selected from the group consisting of Fab, Fab', F(ab') ２ 2. The antibody or antigen-binding portion of claim 1, which is an Fv, scFv, maxibody, minibody, diabody, triabody, tetrabody, or bis-scFv.

15. The antibody or antigen-binding portion of claim 1 , wherein the antibody is monoclonal.

16. The antibody or antigen-binding portion of claim 1 , wherein the antibody is a tetrameric antibody, a tetravalent antibody, or a multispecific antibody.

17. The antibody or antigen-binding portion of claim 1, wherein the antibody is a bispecific antibody that specifically binds to a first antigen and a second antigen, the first antigen being CD122 and the second antigen being other than CD122.

18. An immunoconjugate comprising the antibody or antigen-binding portion of any one of claims 1 to 17 conjugated to a therapeutic agent.

19. 20. The immunoconjugate of claim 18, wherein the therapeutic agent is a cytotoxin, a radioisotope, a chemotherapeutic agent, an immunomodulatory agent, a cytostatic enzyme, a cytolytic enzyme, a therapeutic nucleic acid, an anti-angiogenic agent, an anti-proliferative agent, or a pro-apoptotic agent.

20. A pharmaceutical composition comprising the antibody or antigen-binding portion of any one of claims 1 to 17 and a pharma- ceutically acceptable carrier, diluent or excipient.

21. A pharmaceutical composition comprising the immunoconjugate of claim 18 or 19 and a pharma- ceutically acceptable carrier, diluent or excipient.

22. An antibody or antigen-binding portion according to any one of claims 1 to 17, (a) the amino acid sequence of the VH region; (b) the amino acid sequence of the VL region; or (c) both the amino acid sequences of the VH and VL regions; A nucleic acid molecule encoding

23. 23. An expression vector comprising the nucleic acid molecule of claim 22.

24. 23. A recombinant host cell comprising the nucleic acid molecule of claim 22.

25. A recombinant host cell comprising the expression vector described in claim 23.

26. 1. A method for producing an anti-CD122 antibody or antigen-binding portion thereof, comprising: producing said antibody or antigen binding portion by culturing a recombinant host cell comprising the expression vector of claim 23 under conditions in which said nucleic acid molecule is expressed; isolating the antibody or antigen-binding portion from the host cell or culture medium; The method comprising:

27. A pharmaceutical composition comprising an antibody or antigen-binding portion of any one of claims 1 to 17 for suppressing an immune response in a subject.

28. The pharmaceutical composition of claim 27, wherein the immune response is mediated by CD122.

29. A pharmaceutical composition comprising an antibody or antigen-binding portion of any one of claims 1 to 17 for the treatment or prevention of a disease in a subject.

30. 30. The pharmaceutical composition of claim 29, wherein the disease is an inflammatory disease or an autoimmune disease.

31. 30. The pharmaceutical composition of claim 29, wherein the disease is vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis, or rheumatoid arthritis.

32. 20. A pharmaceutical composition comprising the antibody or antigen-binding portion of any one of claims 1 to 17 for inhibiting IL-15-induced T cell migration from the skin, said pharmaceutical composition being used by contacting said skin.

33. A pharmaceutical composition comprising the immune complex described in claim 18 or 19 for suppressing an immune response in a subject.

34. The pharmaceutical composition described in claim 33, wherein the immune response is mediated by CD122.

35. A pharmaceutical composition comprising the immunoconjugate of claim 18 or 19 for the treatment or prevention of a disease in a subject.

36. The pharmaceutical composition of claim 35, wherein the disease is an inflammatory disease or an autoimmune disease.

37. The pharmaceutical composition of claim 35, wherein the disease is vitiligo, celiac disease, type 1 diabetes, multiple sclerosis, graft-versus-host disease, systemic lupus erythematosus, psoriasis, atopic dermatitis, alopecia areata, ulcerative colitis, or rheumatoid arthritis.

38. A pharmaceutical composition comprising the immune complex described in claim 18 or 19 for inhibiting IL-15-induced T cell migration from the skin, said pharmaceutical composition being used by contacting the skin.

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