How to use anti-IL-2 antibodies

Abstract

Therapeutic uses of modified anti-IL-2 antibodies, including dosages and administration regimens, are described herein. The modified antibodies may confer modified receptor binding specificity to the IL-2-anti-IL2 antibody complex and inhibit IL-2 binding to CD25. The modified anti-IL-2 antibodies may promote the proliferation of a subset of effector immune cells and reduce undesirable effects caused by IL-2. Therapeutic uses of modified anti-IL-2 antibodies include treating cancer, for example, cancers manifesting as solid tumors and their metastases.

Description

[Technical Field]

[0001] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The Sequence Listing in XML format, created on November 10, 2022, is named P-621284-USP_10NOV22.XML and is 97.7 kilobytes in size.

[0002] The present disclosure relates generally to the field of antibodies. In one embodiment, the present disclosure describes the generation and use of modified anti-IL-2 antibodies that may confer modified receptor binding specificity for IL-2. In certain embodiments, the present disclosure describes methods of treating cancer, including cancers that manifest as solid tumors. [Background technology]

[0003] Interleukin-2 (IL-2) is a 15.4 kDa type I cytokine with a four-helical bundle structure. Since its discovery over 30 years ago, the importance of IL-2 in regulating the immune system has been demonstrated many times. IL-2 is primarily produced and secreted by antigen-activated CD4+ T cells. To a lesser extent, IL-2 is also produced by CD8+ T cells, natural killer (NK) cells, dendritic cells, and mast cells.

[0004] IL-2 signaling has two opposing effects. IL-2 can enhance immune responses by activating effector cells and inducing their proliferation. Alternatively, IL-2 can attenuate immune responses by activating and expanding CD4+ regulatory T (Treg) cells. To promote these functions, IL-2 mediates its effects by binding to two forms of IL-2 receptor: i) a trimeric receptor composed of the IL-2Rα (CD25), IL-2Rβ (CD122), and common IL-2Rγ (γc, CD132) chains, or ii) a dimeric receptor consisting only of the IL-2Rβ and IL-2Rγ subunits. Both the dimeric and trimeric receptors can transduce IL-2 binding signaling through the STAT5 pathway. However, IL-2 binds 100-fold more strongly to the αβγ receptor trimer than to the βγ receptor dimer. It has been demonstrated that the binding affinity of hIL-2 for the αβγ trimer is approximately 10 pM, whereas the affinity of hIL-2 for the βγ dimer is 1 nM.

[0005] The difference in affinity for the dimeric and trimeric forms of the IL-2 receptor is one of the important mechanisms responsible for maintaining immunological homeostasis in vivo. Activation of the trimeric receptor is associated with FoxP3-mediated transcription in Tregs, which express a higher number of αβγ trimeric IL-2 receptors on their membranes. In contrast, binding of IL-2 to βγ dimers is associated with activation of NK cells and memory phenotype (MP) CD8+ cells, which express relatively high levels of βγ dimers and very low levels of αβγ trimeric IL-2 receptors. Under normal physiological conditions, natural levels of IL-2 are relatively low, so its primary function appears to be to promote immune tolerance by acting as a Treg activation and proliferation factor. On the other hand, upon immune system activation, IL-2 levels increase, and IL-2 can then bind to βγ dimers and promote the activation and proliferation of memory phenotype effector T cells (MP) CD8+ and NK cells.

[0006] Since the early 1990s, high-dose IL-2 therapy has been used to treat melanoma and metastatic renal cell carcinoma, with response rates ranging from 10% to 15%. While this approach is effective, IL-2-dependent adverse effects, such as potentially fatal vascular leak syndrome (VLS), preclude many patients from consideration for this therapy, and it has not been adopted for other cancers. The short half-life of administered IL-2 necessitates very frequent administration, leading to repeated spikes in circulating IL-2 levels, thereby exacerbating adverse effects. Finally, wild-type IL-2 is not selective and may also enhance the undesired activation of Treg cells.

[0007] It has been discovered that certain antibodies can bind IL-2 and modulate its binding to the βγ dimeric IL-2 receptor or the αβγ trimeric IL-2 receptor. IL-2 complexed with these antibodies has a relatively long half-life, and these IL-2 complexes may activate specific subsets of effector or immune cells. For example, antibody S4B6-mouse IL-2 complexes preferentially activate mouse effector cells in vivo, whereas antibody JES6.1-mouse IL-2 complexes preferentially activate mouse T regulatory cells in vivo. The mechanism of modulation by the JES-6.1 antibody has been elucidated. The JES6.1-mIL-2 complex has been shown to bind to CD25 but not CD122 in vitro.

[0008] Exogenous IL-2 therapy, and even "non-alpha" therapy that does not bind to the CD25 alpha subunit of the IL-2 trimeric receptor, results in the production of endogenous IL-2. The newly secreted endogenous IL-2 preferentially binds to the trimeric IL-2 receptor on Tregs, leading to the proliferation of immunosuppressive Treg cells through a negative feedback loop.

[0009] Increased IL-2 has been implicated as playing a role in viral infection. SARS-CoV-2 is a positive-strand RNA virus in the respiratory coronavirus family (Coronaviridae). The virus enters the host by binding to angiotensin-converting enzyme 2 (ACE2) on lung and gastrointestinal tissues. The course of infection is characterized by an incubation period of approximately 7–14 days, followed by symptoms of dry cough, fever, and shortness of breath. Up to 20% of symptomatic individuals develop severe symptoms, and an average of 3% of cases are fatal due to pulmonary failure. Previous studies on members of the coronavirus family have demonstrated that coronavirus infection leads to an increase in regulatory T lymphocytes, which likely contributes to delayed viral clearance. More recent studies on COVID-19 patients have shown that ICU patients have higher levels of IL-2, IL-7, IL-10, GSCF, IP10, MCP1, MIP1A, and TNF-α than non-ICU patients, suggesting a role for immunopathology in severe disease. Investigations of direct evidence of altered leukocyte homeostasis using immunological characterization of peripheral blood leukocytes from patients infected with SARS-CoV2 indicate that in COVID-19, as in some chronic infections, damage to CD4+ T cell function promotes the excessive activation and possibly subsequent depletion of CD8+ T cells. These perturbations of T cell subsets may ultimately diminish host antiviral immunity. Therefore, therapies that slow viral replication or enhance immune responses to eliminate viral load while reducing some of the associated immunopathology would be highly beneficial.

[0010] The immune response to viruses consists of both the innate and adaptive arms of the immune system. The innate system senses viral RNA / DNA using toll-like receptors (TLRs) and retinoic acid-inducible gene I (RIG-I) proteins and induces an early response. This response includes the production of antiviral cytokines (e.g., interferon-α), chemokines that direct the immune system to the site of infection, and the recruitment of macrophages / dendritic cells. Natural killer (NK) cells (innate lymphocytes) directly kill virus-infected cells in the absence of MHC class I expression. This can occur even if the virus interferes with the MHC class I presentation system.

[0011] Furthermore, during infection responses, NK cells produce interferon-γ (IFN-γ), thereby increasing the expression of MHC class I on cells and enhancing the adaptive immune system's ability to respond. The adaptive immune system consists of T cells (CD4 and CD8) and B cells. CD4+ T cells recognize viral antigens in the context of MHC-II on antigen-presenting cells, amplifying the immune response (via cytokines) and inducing B cell class switching and subsequent production of antiviral antibodies. Activation of CD4 cells, particularly Th1 cells, also releases IFN-γ, thus enhancing viral antigen presentation. CD8+ T cells exert a direct lytic effect on virus-infected cells presenting viral peptides in the context of MHC-I. The initial induction phase of the immune response typically takes 7–10 days to expand the T cell population and generate the cells necessary to eliminate the virus.

[0012] IL-2 is a key mediator in the proliferation and activation of T cells and NK cells. IL-2 is generally believed to play a major role in the secondary signaling required for T cell activation. Expression of the dimeric (βγ) and trimeric (αβγ) IL-2 receptor complexes exhibits lineage selectivity in that the trimeric receptor containing CD25 (α subunit) is highly expressed on regulatory T cells and subsets of activated, short-lived cytotoxic effector T cells, whereas the dimeric receptor is found on naive T cells, memory T cells, and NK cells. As a result, naive T cells, memory T cells, and NK cells can receive signaling through IL-2 binding to the dimeric receptor. Regulatory T cells rely on the high-affinity trimeric receptor complex to enhance their functions, including sequestering IL-2 from binding to memory and naive T cells, thereby reducing the function of these cell populations. The mechanism of action of IL-2 is described in Figure 1.

[0013] Effector T cell subsets also express the trimeric IL-2 receptor complex. Although these cells are highly active, IL-2 binding to these subsets induces activation-induced cell death (AICD). Furthermore, CD25 has been shown to be expressed on pulmonary and vascular endothelium. This expression correlated with pulmonary edema and vascular leakage in a mouse model using high-dose IL-2. It has been suggested that CD25 expression on lung cells is the reason for the pulmonary toxicity of high-dose IL-2 therapy. Furthermore, while pulmonary endothelial cells express CD25 under steady-state conditions, the expression level of CD25 on these cells increased in vivo after IL-2 injection into mice. It has been shown that IL-2-induced pulmonary edema and vascular leak syndrome can be prevented by knocking out CD25 on non-immune cells or by blocking the CD25-binding epitope of IL-2 by using an immunoconjugate of IL-2 with an anti-IL-2 antibody (IL-2 / mAb). It was also demonstrated that in mice genetically modified to lack T and B cells and irradiated sublethally to eliminate remaining immune cells (NK, monocytes, DCs, and granulocytes), the addition of high doses of IL-2 resulted in significant pulmonary edema, indicating a non-immune component.

[0014] Much research has explored the dual role of IL-2 in the lung's ability to clear viral infections. IL-2 has been demonstrated to be necessary for CD8+ T cell proliferation for viral clearance. IL-2 has also been shown to mediate pulmonary edema. For example, in a murine influenza model of influenza virus pulmonary infection, memory CD4+ T cells have been shown to produce high levels of IL-2, and the presence of this IL-2 exacerbates the disease. Regulatory T cells are important for mitigating pathological damage to lung tissue during viral infection. One mechanism by which Tregs control CD8+ effector cells has been hypothesized and demonstrated to be through high-affinity consumption of IL-2 via the CD25 trimeric receptor on Tregs. This effectively removes IL-2 from proliferating effector cells, subsequently limiting their availability and potentially reducing viral clearance. Tregs may also limit the effect of IL-2 on the pulmonary endothelium by sequestering IL-2 from CD25+ endothelial cells. The outcome may depend on the Teff / Treg ratio. High levels of Teff (effector T cells) can lead to viral clearance, but also to excessive levels of IL-2 secreted by immune-activated cells, which can lead to pulmonary edema. In contrast, high Treg proliferation can alleviate the pathology of pulmonary edema, but also reduces viral clearance and can lead to prolonged viral infection.

[0015] Recent data from COVID-19 patients suggest that higher viral loads result in poorer outcomes. Therefore, reduced viral clearance would be associated with poorer outcomes. The role of Tregs in reduced viral clearance was demonstrated using a mouse model of influenza A virus (IAV) infection, in which mice infected with IAV showed higher levels of Tregs in the lungs, spleen, and lymph nodes, along with higher viral loads in lung tissue. This was observed even 6 weeks after the onset of infection. It was suggested that influenza A induces Treg proliferation to evade clearance by the immune response. To evaluate whether an enhanced immune response could increase the clearance of IAV infection in the lungs, researchers used mice previously infected with IAV and then infected them with lymphocytic choriomeningitis virus (LCMV), which elicits a vigorous cytotoxic T lymphocyte response. The extensive immune response in IAV-infected lungs also resulted in pulmonary edema and extensive lung tissue damage. Treatment of IAV-bearing mice with anti-CD25 blocking antibodies prior to LCMV challenge protected them from severe pulmonary edema. These data demonstrate that enhancing immune responses in a setting where Tregs delay viral clearance can induce viral clearance. Furthermore, blocking IL-2 binding to CD25+ cells has been demonstrated to reduce the risk of immune-mediated pulmonary edema during viral clearance.

[0016] IL-2 administered as monotherapy has been shown to enhance antiviral immune responses. By examining the effects of IL-2 therapy during the proliferation, contraction, and memory phases of T cells in LCMV-infected mice, it was demonstrated that IL-2 treatment during the proliferation phase was detrimental to the survival of rapidly dividing effector T cells that transiently upregulated CD25 expression. These effector T cells then directed AICD. In contrast, IL-2 therapy was highly beneficial during the contraction phase, resulting in the survival and activation of virus-specific T cells. IL-2 treatment was also observed to enhance the activation and proliferation of resting memory T cells. However, IL-2 therapy has drawbacks. Due to the short half-life of IL-2, multiple administrations, e.g., daily loading doses followed by weekly administrations, are required, leading to additional associated adverse events and increased risk of immunogenicity. Furthermore, administration of exogenous high-dose IL-2 would be expected to bind to CD25-positive endothelial cells. Indeed, pulmonary edema and vascular leak syndrome are the major serious adverse events of high-dose IL-2 therapy in oncology. The development of technologies to overcome these limitations is crucial for the use of IL-2 as a therapy.

[0017] Those skilled in the art will recognize that the principles discussed above with respect to IL-2 and the treatment of viral infections apply equally to IL-2 and the treatment of bacterial infections or the treatment of cancer.

[0018] Advances in the field of biomolecular engineering provide researchers with unprecedented opportunities to apply molecular design strategies to modify naturally occurring proteins and generate new molecules for targeted disease treatment. In one area, the development of immunotherapeutic agents, such as cytokine- or antibody-based drugs, is being powered by evolving techniques and insights from protein engineering. Thus, there is a need to develop modified anti-IL-2 antibodies that would be used to modulate the function of IL-2 in certain disease states, such as, but not limited to, viral or bacterial infections, and cancer. Summary of the Invention

[0019] In one aspect, a method of treating cancer in a subject comprises administering to the subject a composition comprising an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the cancer in the subject.

[0020] In another aspect, there is provided a method of treating cancer in a subject, comprising administering to the subject an anti-IL-2 antibody and IL-2, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the IL-2 has an amino acid sequence of:

[0021] In another aspect, there is provided a method of treating cancer in a subject, comprising administering to the subject an anti-IL-2 antibody, IL-2, and a checkpoint inhibitor, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the IL-2 is administered by subcutaneous injection, the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, and the checkpoint inhibitor comprises PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, thereby treating the cancer in the subject.

[0022] In another aspect, a method of treating a solid tumor in a subject comprises administering to the subject a composition comprising an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the cancer in the subject.

[0023] In a related embodiment of the methods disclosed herein, multiple doses of a composition comprising an anti-IL2 antibody are administered.

[0024] In another related embodiment of the methods disclosed herein, the method further comprises administering at least a single low dose of IL-2, wherein said low dose of IL-2 is about 15 x 10 per kg of said subject. 3 IU ~500×10 3 In a further related embodiment, the administration of IL-2 comprises subcutaneous administration. In a still further related embodiment, the IL-2 is administered before, simultaneously with, or after administration of the anti-IL-2 antibody. In yet another further related embodiment, the IL-2 is administered as multiple doses. In yet another further related embodiment, multiple doses of IL-2 are administered before, simultaneously with, or after administration of the anti-IL-2 antibody, or any combination thereof.

[0025] In another related aspect of the methods disclosed herein, the method further comprises administering a checkpoint inhibitor. In a further related aspect, the checkpoint inhibitor comprises PD-L1, PD-1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1.

[0026] In another related aspect of the presently disclosed methods, the solid cancer includes melanoma, metastatic melanoma, primary and metastatic melanoma, renal cell carcinoma (RCC), non-small cell lung cancer (NSCLC), head and neck cancer, head and neck squamous cell carcinoma (HNSCC), pancreatic cancer, lung cancer, thyroid cancer, bladder cancer, nasopharyngeal cancer, colorectal cancer (CRC), bile duct cancer (cholangiocarcinoma), uterine cancer, cervical cancer, gallbladder cancer, and cutaneous squamous cell carcinoma.

[0027] In yet another related aspect of the methods disclosed herein, the method includes a second line treatment or a third line treatment, or a combination thereof.

[0028] In another related aspect of the methods disclosed herein, treating the subject reduces tumor size, inhibits or reduces tumor growth, or inhibits or reduces metastasis of the tumor, or any combination thereof.

[0029] In yet another related aspect of the methods disclosed herein, the VH and VL are: (a) VH comprises the amino acid sequence of SEQ ID NO: 26 and VL comprises the amino acid sequence of SEQ ID NO: 27; (b) VH comprises the amino acid sequence of SEQ ID NO: 20 and VL comprises the amino acid sequence of SEQ ID NO: 21; (c) the VH comprises the amino acid sequence of SEQ ID NO: 22 and the VL comprises the amino acid sequence of SEQ ID NO: 23; (d) the VH comprises the amino acid sequence of SEQ ID NO: 24 and the VL comprises the amino acid sequence of SEQ ID NO: 25; or (e) the VH comprises the amino acid sequence of SEQ ID NO: 36 and the VL comprises the amino acid sequence of SEQ ID NO: 37; It has the amino acid sequence:

[0030] In a further related embodiment, the antibody comprises an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab')2, minibody, diabody, or triabody. In yet another further related embodiment, the antibody comprises a heavy chain comprising a mutation that reduces binding to an Fcγ receptor. In yet a further related embodiment, the mutation comprises a L234A, L235A mutation. In yet another further related embodiment, (a) if HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67, then the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 72 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 73; (b) if HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49, then the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 68 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 69; and (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO:55, the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO:70 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO:71.

[0031] In another aspect, there is provided a method of treating cancer in a subject, comprising administering to the subject an anti-IL-2 antibody and IL-2, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the IL-2 has an amino acid sequence of:

[0032] In yet another aspect, there is provided a method of treating a solid tumor in a subject, comprising administering to the subject an anti-IL-2 antibody, IL-2, and a checkpoint inhibitor, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the IL-2 is administered by subcutaneous injection, the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, and the checkpoint inhibitor comprises PD-L1, PD-1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, thereby treating the cancer in the subject. In a related aspect, the checkpoint inhibitor comprises a PD-L1 checkpoint inhibitor.

[0033] In another aspect, there is provided a method of treating non-small cell lung cancer (NSCLC) in a subject, comprising administering to the subject a composition comprising an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the NSCLC in the subject.

[0034] In another aspect, there is provided a method of treating non-small cell lung cancer (NSCLC) in a subject, comprising administering to the subject an anti-IL-2 antibody and IL-2, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, and the IL-2 is administered subcutaneously, thereby treating the NSCLC in the subject.

[0035] In another aspect, there is provided a method of treating non-small cell lung cancer (NSCLC) in a subject, comprising administering to the subject an anti-IL-2 antibody, IL-2, and a checkpoint inhibitor, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, the IL-2 is administered subcutaneously, and the checkpoint inhibitor comprises PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, thereby treating the NSCLC in the subject.

[0036] In another aspect, there is provided a method of treating renal cell carcinoma (RCC) in a subject, comprising administering to the subject an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the RCC in the subject.

[0037] In another aspect, there is provided a method of treating melanoma in a subject, comprising administering to the subject a composition comprising an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the melanoma comprises a primary tumor or a metastatic melanoma, or a combination thereof; and the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, wherein the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the melanoma in the subject.

[0038] In another aspect, there is provided a method of treating cancer in a subject, comprising administering to the subject an anti-IL-2 antibody, IL-2, and a checkpoint inhibitor, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, the dose of IL-2 is a low dose, and the checkpoint inhibitor is a PD-L1 checkpoint inhibitor, wherein the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the cancer in the subject.

[0039] In another related embodiment, the method further comprises administering IL-2. In a further related embodiment, administering IL-2 comprises subcutaneous administration. In another further related embodiment, IL-2 is administered as a single dose. In yet another further related embodiment, IL-2 is administered before, simultaneously with, or after administration of said anti-IL-2 antibody. In yet another further related embodiment, IL-2 is administered as multiple doses. In another further related embodiment, multiple doses of IL-2 are administered before, simultaneously with, or after administration of said anti-IL-2 antibody, or any combination thereof. In yet another further related embodiment, the dose of IL-2 is about 15 x 10 per kg of said subject. 3 IU~270×10 3 It's IU.

[0040] In another related aspect, the method further comprises administering a checkpoint inhibitor. In another related aspect, the method further comprises administering IL-2 and a checkpoint inhibitor.

[0041] 11. The method of claim 10, wherein the checkpoint inhibitor comprises PD-L1, PD-1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1. In another further related aspect, the checkpoint is PD-L1. In yet another further related aspect, the PD-L1 checkpoint inhibitor is avelumab.

[0042] In another related aspect, the cancer comprises a solid cancer. In a further related aspect, the solid cancer comprises non-small cell lung cancer (NSCLC), head and neck cancer, head and neck squamous cell carcinoma (HNSCC), pancreatic cancer, lung cancer, thyroid cancer, bladder cancer, nasopharyngeal cancer, melanoma, metastatic melanoma, primary and metastatic melanoma, colorectal cancer (CRC), bladder cancer, bile duct cancer (cholangiocarcinoma), uterine cancer, cervical cancer, gallbladder cancer, or renal cell carcinoma (RCC). In another further related aspect, the solid cancer comprises non-small cell lung cancer (NSCLC), melanoma, metastatic melanoma, primary and metastatic melanoma, or renal cell carcinoma (RCC). In yet another further related aspect, the solid cancer comprises non-small cell lung cancer (NSCLC). In another further related aspect, the non-small cell lung cancer (NSCLC) comprises unresectable advanced or metastatic cancer.

[0043] In a related aspect, the method includes a second line treatment or a third line treatment, or a combination thereof.

[0044] In another related embodiment, the anti-IL2 antibody is administered for about 3 months to 1 year.

[0045] In another related aspect, the method of treating the subject is to reduce tumor size, inhibit or reduce tumor growth, or inhibit or reduce metastasis of the tumor, or any combination thereof.

[0046] In another related aspect, VH and VL are (a) VH comprises the amino acid sequence of SEQ ID NO: 26 and VL comprises the amino acid sequence of SEQ ID NO: 27; (b) VH comprises the amino acid sequence of SEQ ID NO: 20 and VL comprises the amino acid sequence of SEQ ID NO: 21; (c) the VH comprises the amino acid sequence of SEQ ID NO: 22 and the VL comprises the amino acid sequence of SEQ ID NO: 23; (d) the VH comprises the amino acid sequence of SEQ ID NO: 24 and the VL comprises the amino acid sequence of SEQ ID NO: 25; or (e) the VH comprises the amino acid sequence of SEQ ID NO: 36 and the VL comprises the amino acid sequence of SEQ ID NO: 37; It has the amino acid sequence:

[0047] In another related embodiment, the antibody comprises an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab')2, minibody, diabody, or triabody. In a further related embodiment, the antibody comprises a heavy chain comprising a mutation that reduces binding to an Fcγ receptor. In yet a further related embodiment, the mutation comprises an L234A, an L235A mutation.

[0048] In another related aspect, (a) if HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67, then the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 72 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 73; (b) if HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49, then the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 68 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 69; and (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, and LCDR2 YAS and LCDR3 comprises the amino acid sequence of SEQ ID NO:55, the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO:70 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO:71.

[0049] In another related aspect, the undesirable effects caused by IL-2 include activation of regulatory T cells, CD25 + These include one or more of T effector cell apoptosis, IL-2-induced pulmonary edema, IL-2-induced pneumonia, or IL-2-induced vascular leakage. In a further related embodiment, the anti-IL2 antibody binds to IL-2, and said binding reduces or eliminates IL-2 binding to CD25. In yet another further related embodiment, IL-2 binding to CD132 / CD122 is not reduced or inhibited.

[0050] The patent or patent application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0051] The present disclosure of modified anti-IL-2 antibodies, both as to their making and methods of use, together with their objects, features, and advantages, can best be understood by reference to the following detailed description when read in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0052] [Figure 1] FIG. 1 is a schematic diagram of the mechanism of action of IL-2 and its dual role in regulating immune responses.

[0053] [Figure 2] FIG. 1 is a schematic diagram of anti-IL-2 antibody-directed immunotherapy.

[0054] [Figure 3A] Schematic diagram of the progression of COVID-19 infection. [Figure 3B]Schematic diagram of potential anti-IL-2 therapy as an adjuvant intervention. Figure 3A is adapted from Shi Y et al. (2020) COVID-19 infection: the perspectives on immune responses. Cell Death & Differentiation 27, 1451–1454 (doi:10.1038 / s41418-020-0530-3), Figure 1.

[0055] [Figure 4A] FIG. 1 shows a representative SPR sensorgram of the binding of JES6.1 antibody to human IL-2. [Figure 4B] FIG. 1 shows a representative SPR sensorgram of the binding of JES6.1 antibody to mouse IL-2. [Figure 4C] FIG. 1 shows a representative SPR sensorgram of the binding of the JES6.1RMC antibody to human IL-2. [Figure 4D] FIG. 1 shows a representative SPR sensorgram of the binding of JES6.1RMC antibody to mouse IL-2.

[0056] [Figure 5A] FIG. 1 shows the results of IL-2 binding of YSD clones expressing the JES6.1 antibody in scFv format. [Figure 5B] FIG. 1 shows the results of IL-2 binding of YSD clones expressing the JES6.1 antibody in scFv format. [Figure 5C] Figure 5 shows the results of IL-2 binding of YSD clones expressing the JES6.1 antibody in scFv format. Fluorescence levels on the X-axis correspond to the scFv expression level of Jes6.1, and fluorescence levels on the Y-axis correspond to the binding of human or mouse IL-2. Figure 5A shows a negative control without IL-2. Figure 5B shows a JES6.1 YSD clone containing 1000 nM human IL-2. Figure 5C shows a YSD expressing mouse IL-2 incubated with 100 nM labeled JES6.1.

[0057] [Figure 6A]Figure 1 shows binding of isolated yeast surface display clones to IL-2 (0.1 nM). Mean fluorescence intensity (Em 655 nM) was normalized to yeast surface expression levels. Negative YSD clones were labeled with 500 nM hIL-2.

[0058] [Figure 6B] Figure 1 shows nonspecific binding of YSD clones to a mixture of OX40 / PD-1 / TNFR2. Clones were labeled with a 500 nM mixture. A TNFR2-binding yeast clone served as a positive control.

[0059] [Figure 7] Figure 1 shows the purification of BDG17.023 IgG. The antibody was run at 0.5 ml / min on a GE Superdex 200 10 / 300 gradient (CV = 25 ml) in PBS buffer. The main peak (0.38 CV) corresponds to typical aggregates, while the second peak (0.51 CV) with a retention of approximately 12.9 ml is typical of normal human IgG.

[0060] [Figure 8A] FIG. 1 shows the binding kinetics of BDG17.023 IgG to hIL-2. [Figure 8B] FIG. 1 shows the binding kinetics of BDG17.023 IgG to mIL-2.

[0061] [Figure 9] Figure 1 shows receptor discrimination by the BDG17.023-IL-2 complex using SPR response traces. BDG17.023 was immobilized on a CM5 chip, and hIL-2 (60 RU), CD122 (20 RU), and CD25 (0 RU) were streamed as indicated by the arrows.

[0062] Figures 10A-10D show splenic immune cell populations from mice treated with JES6.1-mIL-2 complexes and BDG17.023-hIL-2 complexes. [Figure 10A]FIG. 1 shows the percentage of immune cell populations from mice treated with JES6.1-mIL-2 complexes. [Figure 10B] FIG. 1 shows the memory phenotype effector T cell (MP) CD8+ / Treg ratio in mice treated with JES6.1-mIL-2 complex. [Figure 10C] FIG. 1 shows the percentage of immune cell populations from mice treated with BDG17.023-hIL-2 complexes. [Figure 10D] FIG. 1 shows the MP CD8+ / Treg ratio in mice treated with BDG17.023-hIL-2 complexes.

[0063] [Figure 11] FIG. 1 shows an alignment of the amino acid sequences of the heavy chain variable regions of JES6.1, clone 1 (17.021), clone 2 (17.022), clone 4 (17.023), clone 5 (17.030), and clone 6 (17.035).

[0064] [Figure 12] FIG. 1 shows an alignment of the amino acid sequences of the light chain variable regions of JES6.1, clone 1 (17.021), clone 2 (17.022), clone 4 (17.023), clone 5 (17.030), and clone 6 (17.035).

[0065] [Figure 13A] FIG. 1 shows an alignment of the amino acid sequences of the heavy chain variable regions of humanized clone 17.014, clone 17.038, clone 17.043, clone 17.053, and clone 17.054. [Figure 13B] Figure 1 shows the amino acid sequence alignment of the light chain variable regions of humanized clones 17.014, 17.038, 17.043, 17.053, and 17.054. Black triangles indicate IMGT CDR positions. Bold / italic text indicates ABR / CDR positions, respectively.

[0066] Figures 14A-G. Binding kinetics of the indicated antibodies to human IL-2. [Figure 14A] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.038 to human IL-2. [Figure 14B] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG 17.043 to human IL-2. [Figure 14C] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.053 to human IL-2. [Figure 14D] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.054 to human IL-2. [Figure 14E] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.066 to human IL-2. [Figure 14F] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.067 to human IL-2. [Figure 14G] Figure 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.069 to human IL-2. The binding kinetics of BDG 17.038, BDG 17.043, BDG 17.066, BDG 17.067, and BDG 17.069 were determined by the multi-cycle method. The binding kinetics of BDG 17.053 and BDG 17.054 were determined by the single-cycle method.

[0067] Figures 15A-15B. Binding kinetics of the indicated antibodies to cynomolgus IL-2. [Figure 15A]FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.067 to cynomolgus IL-2. [Figure 15B] FIG. 1 shows surface plasmon resonance (SPR) sensorgram traces of the binding kinetics of anti-IL-2 antibody clone BDG17.069 to cynomolgus IL-2.

[0068] [Figure 16A] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16B] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16C] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16D] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16E] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16F] Figure 1 shows a differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates T onset, the thick green dashed line indicates Tm1, and where applicable, Tm2. [Figure 16G]Figure 16 shows differential scanning fluorimetry (DSF) analysis of the melting points of the indicated IgGs. The light green dashed line indicates the T onset, and the thick green dashed line indicates Tm1 and, where applicable, Tm2. The anti-IL-2 clones analyzed are BDG17.038 (Figure 16A), BDG17.043 (Figure 16B), BDG 17.053 (Figure 16C), BDG 17.054 (Figure 16D), BDG17.066 (Figure 16E), BDG17.067 (Figure 16F), and BDG17.069 (Figure 16G).

[0069] Figures 17A-G show receptor discrimination of the indicated antibody / IL-2 complexes by tracking the SPR response. Antibodies were immobilized on a CM5 chip, and hIL-2, CD122, and CD25 were streamed as indicated by the arrows. [Figure 17A] FIG. 1 shows the general sequence of compound injections onto an SPR chip representing sequential anti-IL-2 antibodies complexed with human IL-2 (hIL2) and binding to CD122 but not CD25. [Figure 17B] FIG. 1 shows SPR response: BDG17.038. [Figure 17C] FIG. 1 shows SPR response: BDG017.043. [Figure 17D] FIG. 1 shows SPR response: BDG17.054. [Figure 17E] FIG. 1 shows SPR response: BDG17.066. [Figure 17F] FIG. 1 shows SPR response: BDG17.067. [Figure 17G] FIG. 1 shows SPR response: BDG17.069 (AU-007).

[0070] Figures 18A and 18B show that anti-human IL-2 antibodies (clones 17.043 and 17.054) exhibit potent immunostimulatory effects in vivo. Anti-IL-2 antibody / hIL-2 complexes expand effector cell populations without any observed effect on regulatory T cells. [Figure 18A]FIG. 1 shows that C57BL / 6 mice were given anti-IL-2 antibody (10 ug) pre-complexed with 0.5 ug hIL-2 daily for 4 days. [Figure 18B] C57BL / 6 mice were administered anti-IL-2 antibody (25 μg) pre-complexed with 1.25 μg hIL-2 daily for 4 days. On day 5, splenocytes were isolated and immune cell populations were analyzed by flow cytometry. Mean values ​​for each experimental group are shown (n=6 / group). Lymphocytes were gated according to side scatter and forward scatter parameters, and subsequent immune cell subpopulations were gated as follows: Tregs (CD45+, CD3+, CD4+, CD25+, FoxP3+), CD8 T cells (CD45+, CD3+, CD8+, CD122+, CD25-), NKT cells (CD45+, CD3+, CD49b+, NK1.1+), and NK cells (CD45+, CD3-, CD49b+, NK1.1+).

[0071] Figures 19A and 19B show that anti-human IL-2 antibodies (clones 17.043 and 17.054) exhibit potent dose-dependent immunostimulatory effects in vivo. [Figure 19A] C57BL / 6 healthy mice were administered anti-IL-2 antibody / hIL-2 complexes (25ug / 1.25ug, respectively) daily for 4 days. On day 5, splenocytes were isolated and immune cell populations were analyzed by flow cytometry. [Figure 19B]This figure shows that anti-human IL-2 antibodies exhibit potent in vivo immunostimulatory effects in a dose-dependent manner. Healthy C57BL / 6 mice were administered increasing doses of anti-IL-2 antibody / hIL-2 complexes daily as indicated. On day 5, splenocytes were isolated and immune cell populations were analyzed using flow cytometry. Lymphocytes were gated according to side scatter and forward scatter parameters, and subsequent immune cell subpopulations were gated as follows: Tregs (CD45+, CD3+, CD4+, CD25+, FoxP3+), CD8 T cells (CD45+, CD3+, CD8+, CD122+, CD25-), NKT cells (CD45+, CD3+, CD49b+, NK1.1+), and NK cells (CD45+, CD3-, CD49b+, NK1.1+).

[0072] Figures 20A and 20B show that anti-human IL-2 antibodies (clones 17.043 and 17.054) exhibit a safe dosing regimen in vivo. [Figure 20A] FIG. 1 shows that healthy C57BL / 6 mice were given daily doses of anti-IL-2 antibody / hIL-2 complexes (10 ug / 0.5 ug, respectively) for 4 days. [Figure 20B] C57BL / 6 healthy mice were administered anti-IL-2 antibody / hIL-2 complexes (25 μg / 1.25 μg, respectively) daily for 4 days. At the end of the experiment, mice were weighed and the percent weight change was calculated relative to each mouse's weight at the start of the study. The mean percent body weight (BW) change for each experimental group (n=6 / group) is shown.

[0073] Figures 21A and 21B show the results of mean tumor volume. Anti-IL-2 antibodies (clones 17.038, 17.043, 17.053, and 17.054) inhibit tumor growth in an I / O-resistant tumor model with an acceptable safety profile. C57BL / 6 healthy mice were inoculated with B16F10 melanoma tumor cells on day 0. On day 5, mice were randomized into experimental groups (n = 10 / group) and administered anti-IL-2 antibody / hIL-2 complexes (20 μg / 1 μg, respectively) or PBS daily for 4 days. 17.054 is the parent antibody of 17.069 (AU-007), but lacks the LALA effector-silent mutation in the Fc domain engineered into AU-007. [Figure 21A] FIG. 1 shows changes in tumor volume in each experimental group. [Figure 21B] Figure 17. Change in body weight for experimental groups 17.043 and 17.054. The percentage of body weight change was calculated relative to the weight of each mouse at the start of the study.

[0074] Figures 22A-22G show the results of analyzing different formulations of the anti-IL-2 antibody clone BDG 17.069. [Figure 22A] FIG. 1 shows BDG 17.069 parameters at T=0. [Figure 22B] FIG. 1 shows the appearance, pH, protein concentration, and sub-visual particle formation of BDG 17.069 after incubation at T=0 and 1 and 2 weeks at 40° C. [Figure 22C] FIG. 1 shows BDG 17.069 SEC, caliper-SDS and capillary isoelectric focusing analysis after T=0 and 1 and 2 weeks of incubation at 40° C. [Figure 22D] FIG. 1 shows the appearance, pH, protein concentration, and subvisual particle formation of BDG 17.069 at T=0 and after 3 days of stirring at 300 rpm. [Figure 22E]FIG. 1 shows BDG 17.069 SEC, caliper-SDS and capillary isoelectric focusing analysis at T=0 and after 3 days of stirring at 300 rpm. [Figure 22F] FIG. 1 shows the appearance, pH, protein concentration, and subparticle formation of BDG 17.069 at T=0 and after 5 cycles of freeze / thaw. [Figure 22G] FIG. 1 shows BDG 17.069 SEC, caliper-SDS, and capillary isoelectric focusing analyses at T=0 and after 5 cycles of freeze / thaw.

[0075] [Figure 23] FIG. 10 graphically depicts the dosing and administration scheme of BDG17.069 Q2w monotherapy (left-A), combination BDG17.069+IL-2 loading dose (center-B), and combination BDG17.069 Q2w+IL-2 Q2w (right-C).

[0076] [Figure 24] Figure 1 shows details regarding treatment duration and efficacy. Patients in Cohort 1 received 0.5 mg / kg BDG 17.069 and patients in Cohort 2 received 1.5 mg / kg BDG 17.069.

[0077] [Figure 25] FIG. 1 shows pharmacokinetic data showing the initial pharmacokinetic profile of the first three patients receiving monotherapy BGD 17.096.

[0078] Figures 26A-26D graphically depict pharmacodynamic (PD) data demonstrating the efficacy and mechanism of action of BDG 17.069. [Figure 26A] Figure 1 shows the % change in CD4+ Treg population. [Figure 26B] FIG. 26 shows the CD8 / Treg ratio and includes an explanation of the symbols in FIGS. 26A and 26B. [Figure 26C] FIG. 1 shows total circulating IFN-γ. [Figure 26D]FIG. 26 shows eosinophil counts and legends for FIGS. 26C-26D (circles - patient 1; triangles - patient 2; squares - patient 3).

[0079] 27A to 27H are BDG Technical characteristics of the 17.069 (AU-007) antibody, including a schematic of the injection schedule with SPR sensorgram traces. Figures 27A and 27B-D show that AU-007 binds human IL-2 with high affinity and inhibits binding to hCD25 while maintaining binding to hCD122. The affinity and epitope binding site were assessed using surface plasmon resonance (SPR). [Figure 27A] FIG. 1 shows SPR sensorgram traces and calculated binding kinetics of AU-007 bound to a CM5 chip with hIL-2 serving as the analyte. [Figure 27B] FIG. 1 shows an exemplary model of IL-2 binding as part of the human IL-2-trimeric receptor complex compared to AU-007 (yellow), which blocks IL-2 access to CD25. [Figure 27C] FIG. 1 shows AU-007 epitope binding analysis. [Figure 27D] Figure 27E-H: AU-007 epitope binding analysis. AU-007 was captured on a CM5 chip and soluble hIL-2 was injected to form a complex. Soluble hCD25 was then injected, followed by soluble hCD122. The figure shows SPR sensorgram traces of Ab / IL-2 / IL-2R complex formation. Arrows indicate where hIL-2, hCD25, and hCD122 were injected. Figures 27E-H: naIL-2 (hIL-2 / hCD25 conjugate) inhibits binding to hCD25 while maintaining binding to hCD122. [Figure 27E] FIG. 1 is a schematic diagram showing that biotinylated hCD25 was captured on a CM5 chip, followed by injection of soluble hIL-2, soluble hCD25, and naIL-1. [Figure 27F]FIG. 1 is a schematic diagram showing that biotinylated hCD25 was captured on a CM5 chip, followed by injection of soluble hIL-2, soluble hCD25, and naIL-1. [Figure 27G] FIG. 1 is a schematic diagram showing that Fc-tagged hCD122 was captured on a CM5 chip, followed by injection of soluble hIL-2, soluble hCD25, and soluble naIL-2. [Figure 27H]

[0033] Figure 1 is a schematic diagram showing that Fc-tagged hCD122 was captured on a CM5 chip, followed by injection of soluble hIL-2, soluble hCD25, and soluble naIL-2. The figure shows SPR sensorgram traces of IL-2R / cytokine complex formation. Arrows indicate where hIL-2, hCD25, and hCD122 were injected.

[0080] Figures 28A-28H show that AU-007, but not HD IL-2 or naIL-2, can capture and redirect endogenous IL-2 in hPBMCs to disrupt the autoinhibitory loop. AU-007 promotes NK and CD8 T cell proliferation while completely inhibiting regulatory T cell proliferation. Figures 28A-28E (keywords in Figure 28A): Naive hPBMCs were treated once on day 0 with 1 nM naIL-2 (purple) or HD IL-2 (1 nM) in combination with 1 μM isotype control Ab (black), 1 μM AU-007 (red), or 10 μM AU-007 (turquoise). Cultures were monitored for 7 days, and immune cell subpopulations were analyzed daily by flow cytometry. Values ​​were normalized to the untreated sample (UNT) for each day. naIL-2, like AU-007, promotes NK proliferation, but is unable to inhibit Treg proliferation. [Figure 28A] FIG. 1 shows that AU-007 completely inhibits Treg expansion in culture. [Figure 28B] FIG. 1 shows that AU-007 significantly increases the Teff:Treg ratio. [Figure 28C] FIG. 10 shows that AU-007 does not interfere with NK. [Figure 28D]FIG. 28B shows that AU-007 downregulates suppressive markers of CD4+ Tregs in FIG. 28A, as defined by a significant decrease in MFI of CD25. [Figure 28E] FIG. 28B shows that AU-007 downregulates suppressive markers of CD4+ Tregs in FIG. 28A, as defined by a significant decrease in MFI of FoxP3. [Figure 28F] FIG. 1 shows that AU-007 rescues activated lymphocyte viability reduced by treatment with HD IL-2. [Figure 28G] FIG. 1 shows that AU-007 rescues activated lymphocyte viability reduced by treatment with HD IL-2. [Figure 28H] 28A-28C show that AU-007 rescues activated lymphocyte viability reduced by treatment with HD IL-2. Keyword: Figure 28H. hPBMC cultures were stimulated once with anti-CD3 / anti-CD28 Ab with or without 10 μM AU-007. Three days after stimulation, all samples received HD IL-2 (1 nM) and were monitored daily for cell viability using flow cytometry.

[0081] Figures 29A-29K show that AU-007 binds to endogenous IL-2 in human PBMCs and disrupts the negative feedback loop. Figures 29A-29E (keywords in Figure 29E): Naive hPBMCs were treated once on day 0 with either 1 uM AU-007 (red) or an isotype control antibody (blue). No exogenous IL-2 was added. Cultures were monitored for 7 days, and immune cell subpopulations were analyzed daily by flow cytometry. Values ​​were normalized to the untreated sample (UNT) for each day. [Figure 29A] FIG. 1 shows that AU-007 completely inhibits Treg proliferation. [Figure 29B] FIG. 1 shows that AU-007 significantly increases the Teff:Treg ratio. [Figure 29C] FIG. 10 shows that AU-007 does not interfere with NK. [Figure 29D] FIG. 29B shows that AU-007 downregulates suppressive markers of CD4+ Tregs in FIG. 29A, as defined by a significant decrease in CD25 mean fluorescence intensity (MFI). [Figure 29E] FIG. 29B shows that AU-007 downregulates suppressive markers of CD4+ Tregs in FIG. 29A, as defined by a significant decrease in mean fluorescence intensity (MFI) of FoxP3. [Figure 29F] Figure 1 shows that total hPBMCs were stimulated for 24 hours with anti-CD3 / anti-CD28 (stimulation only, green), or stimulated with anti-CD3 / anti-CD28 in the presence of 200 nM AU-007 mAb (red), or stimulated with 200 nM isotype control mAb (blue). [Figure 29G] Figure 1 shows that total hPBMCs were stimulated for 24 hours with anti-CD3 / anti-CD28 (stimulation only, green), or stimulated with anti-CD3 / anti-CD28 in the presence of 200 nM AU-007 mAb (red), or stimulated with 200 nM isotype control mAb (blue). [Figure 29H] Figure 1 shows that total hPBMCs were stimulated for 24 hours with anti-CD3 / anti-CD28 (stimulation only, green), or stimulated with anti-CD3 / anti-CD28 in the presence of 200 nM AU-007 mAb (red), or stimulated with 200 nM isotype control mAb (blue). [Figure 29I] Figure 29 shows that total hPBMCs were stimulated for 24 hours with anti-CD3 / anti-CD28 (stimulation only, green), or with anti-CD3 / anti-CD28 in the presence of 200 nM AU-007 mAb (red), or with 200 nM isotype control mAb (blue). No exogenous IL-2 was added. Immune cell subpopulations were analyzed by flow cytometry. AU-007 inhibits Tregs without interfering with effector cells and NKs (Figures 29F-I). [Figure 29J] FIG. 10 shows that AU-007 downregulates suppressive markers of CD4+ Tregs from panel G, as defined by a significant decrease in MFI of CD25. [Figure 29K]FIG. 10 shows that AU-007 downregulates suppressive markers of CD4+ Tregs from panel G, as defined by a significant decrease in MFI of FoxP3.

[0082] Figures 30A-30C show that AU-007 and naIL-2 do not interfere with CD122 / CD132-STAT5 signaling activity. IL-2 / IL-2 dimeric receptor signaling was detected using the HEK239-dimer-STAT5-SEAP reporter cell line, which stably expresses the human IL-2 dimeric receptor (CD122 / CD132) without CD25 expression and drives the expression of secreted placental alkaline phosphatase (SEAP) under the STAT5 promoter. [Figure 30A] FIG. 1 shows that the expression levels of CD25, CD122, and CD132 were detected using flow cytometry verifying the exclusive expression of dimeric receptors. [Figure 30B] FIG. 1 shows dose-response curves for IL-2 alone (red circles), or in the presence of 200 nM AU-007 (blue squares), or in the presence of 200 nM anti-IL-2 antibody that blocks the interaction with the dimeric receptor (green triangles). [Figure 30C] Figure 30 shows dose-response curves for naIL-2 (filled circles). HEK239-dimer-STAT5-SEAP reporter cells were treated with increasing concentrations of hIL-2 alone or the indicated anti-hIL-2 antibodies (Figure 30B) or increasing concentrations of naIL-2 (Figure 30C). Cumulative levels of SEAP were measured from the cell culture medium 24 hours after treatment, and functional EC-50 values ​​were calculated using GraphPad (Figures 30B-C).

[0083] Figures 31A-31G show that AU-007 captures endogenous IL-2 and exhibits potent immunostimulatory effects in vivo even 8 days after a single treatment. NOG-EXL mice were transplanted with hPBMCs from three human donors to examine donor-to-donor variability. Ten days after hPBMC engraftment, mice were randomized into test groups (9 mice per cohort, each consisting of 3 mice from each donor; colored dots represent the average for each donor across all cohorts). Mice in the test groups were treated once with 20 mg / kg of AU-007 or 20 mg / kg of isotype control Ab (hIgG1-LALA) without the addition of exogenous IL-2. [Figure 31A] FIG. 1 is a diagram showing an experimental outline. [Figure 31B] FIG. 1 shows detection of AU-007 / hIL-2 complexes from mouse serum using ELISA, n=9±SD. [Figure 31C] FIG. 1 shows immune cell analysis of splenocytes using flow cytometry, n=9±SE. [Figure 31D] FIG. 1 shows immune cell analysis of splenocytes using flow cytometry, n=9±SE. [Figure 31E] FIG. 1 shows immune cell analysis of splenocytes using flow cytometry, n=9±SE. [Figure 31F] FIG. 1 shows immune cell analysis of splenocytes using flow cytometry, n=9±SE. [Figure 31G] Figure 31 shows immune cell analysis of splenocytes using flow cytometry, n=9±SE. Human immune cells were defined as hCD45+hCD3+ from total lymphocytes, and subpopulations were defined using anti-human antibodies as indicated. Statistical analysis was performed using a two-way ANOVA test; *p<0.05, ****p<0.0001. (Keywords below Figures 31F-G)

[0084] Figures 32A-C show that the IL-2 negative feedback loop triggered by endogenous IL-2 limits the activity of modified IL-2-based therapy. [Figure 32A] Schematic diagram of the role of IL-2 as an immunoregulator in homeostasis and inflammation. The header of Figure 32A also indicates the status / function of Figures 32B and 32C. [Figure 32B] FIG. 1 shows that exogenous administration of modified IL-2 with biased selectivity for dimer-expressing cells promotes proliferation of CD25-negative (CD25−) effector cells, but is impaired by endogenous IL-2, which pushes the system back toward homeostasis. [Figure 32C] FIG. 1 shows that AU-007 captures and redirects endogenous IL-2, breaking the autoinhibitory loop and allowing CD25-negative (CD25−) effector cells to expand, expanding the inflammatory and immune stimulatory phase.

[0085] [Figure 33A] FIG. 1 shows HEK-293 cells expressing the IL-2 dimeric receptor were incubated with IL-2 (red) or IL2 plus AU-007 (17.069) (blue) or a control antibody with known dimer-inhibiting properties (green). [Figure 33B] HEK-293 cells expressing the IL-2 dimeric receptor were incubated with IL-2 (red) or IL-2 plus AU-007 (17.069) (blue) or a control antibody with known dimer-inhibiting properties (green). The output is the production of secreted placental alkaline phosphatase (SEAP) after stimulation. SEAP was detected using a phosphatase substrate, and absorbance was measured at 620 nm. AU-007 did not inhibit the ability of IL-2 to stimulate the IL-2 dimeric receptor on HEK293 cells (EC50 = 0.71 pM for IL-2 alone and 0.78 pM in the presence of AU-007).

[0086] [Figure 34A] FIG. 1 shows the results of a pSTAT5 IC50 assay in human peripheral blood mononuclear cells (PBMCs). [Figure 34B] FIG. 1 shows the results of a pSTAT5 IC50 assay in human peripheral blood mononuclear cells (PBMCs). [Figure 34C]FIG. 1 shows the results of a pSTAT5 IC50 assay in human peripheral blood mononuclear cells (PBMCs). [Figure 34D] Figure 34 shows the results of a pSTAT5 IC50 assay in human peripheral blood mononuclear cells (PBMCs). Human PBMCs were incubated with IL-2 in the presence (blue line) or absence (black line) of AU-007. The data demonstrate that AU-007 inhibits the ability of IL-2 to induce STAT5 phosphorylation in IL-2 trimeric receptor-expressing regulatory T cells (CD3+CD4+CD25+CD127-FoxP3+) (Figure 34A), but not in dimeric receptor-expressing CD3-CD56+ NK cells (Figure 34B), memory phenotype (MP) CD8+ T cells (Figure 34C), or CD3+CD8+CD56+ NKT cells (Figure 34D).

[0087] [Figure 35A] FIG. 1 shows the percent cells (%) from parent of CD8 MP Teff cells after administration of AU-007 / hIL-2. [Figure 35B] FIG. 1 shows the percent cells (%) from the parent of NK cells after administration of AU-007 / hIL-2. [Figure 35C] FIG. 1 shows the percent cells (%) from the parent of NKT cells after administration of AU-007 / hIL-2. [Figure 35D] FIG. 1 shows the percent cells (%) from parent of CD4+ Treg cells after administration of AU-007 / hIL-2. [Figure 35E] Figure 1 shows the percent (%) of parental CD8+ Treg cells after administration of AU-007 / hIL-2. Briefly, C57B1 / 6 mice were injected daily with AU-007, IL-2 + AU007, or IL2 + control antibody for 4 days, and spleens were harvested on day 5. Splenocytes were phenotyped for the percentage of lymphocyte populations. AU-007 significantly expanded CD8+ memory cells, NK cells, and NKT cells, but not regulatory T cells.

[0088] [Figure 36A]Figure 1 shows the percent cells (%) from parent of CD8 MP Teff cells after administration of BDG17.054 / hIL-2. [Figure 36B] FIG. 1 shows the percent cells (%) from parent of NK cells after administration of BDG17.054 / hIL-2. [Figure 36C] FIG. 1 shows the percent cells (%) from the parent of NKT cells after administration of BDG17.054 / hIL-2.

[0089] [Figure 37A]

[0023] Figure 1 presents data showing that administration of BDG17.069 plus human IL-2 to MC38 colon cancer-bearing mice induces regression and, in the presence of anti-PD-1, tumor eradication. [Figure 37B]

[0023] Figure 1 presents data showing that administration of BDG17.069 plus human IL-2 to MC38 colon cancer-bearing mice induces regression and, in the presence of anti-PD-L1, tumor eradication.

[0090] [Figure 38A] FIG. 1 presents data showing that administration of BDG17.069 plus human IL-2 inhibits tumor growth in an LL / 2 (lung) cancer model. [Figure 38B]

[0023] Figure 1 presents data showing that administration of BDG17.069 plus human IL-2 inhibits tumor growth in an LL / 2 (lung) cancer model. BDG17.069 (AU-007) reduced LL / 2 tumor growth by 74% compared to saline control. Note that AU-007 is administered with human IL-2 because AU-007 does not bind to mouse IL-2.

[0091] [Figure 39] Schematic of the updated Phase 1 dose escalation regime. The solid green border indicates work performed and / or in progress. The dashed green line indicates progression to the next step (as of November 2023).

[0092] [Figure 40]AU-007 monotherapy: (Arm 1A) Treatment duration and best response (as of October 13, 2023).

[0093] [Figure 41A] FIG. 1 shows AU-007+IL-2 (Arm 1B (AU-007+single loading dose of aldesleukin)) treatment duration and best response (as of October 2023). [Figure 41B] FIG. 1 shows the treatment period and best response (as of October 2023) for AU-007 + IL-2 (Arm 1C (AU-007 + aldesleukin (Q2W))).

[0094] Figures 42A, 42B, and 42C present safety data. [Figure 42A] FIG. 1 shows Arm 1A (first four cohorts) AU-007 monotherapy safety data: mild toxicity profile. [Figure 42B] Population statistics (as of October 13, 2023) for all three arms (1A, 1B, and 1C) are shown. *All grade 3 / 4 drug-related AEs were transient (3-7 days) lymphopenia. **A single drug-related SAE of transient (approximately 12 hours) grade 2 CRS occurred in a patient with cutaneous squamous cell carcinoma who received AU-007 + Q2W 135K IU / kg aldesleukin. The patient became symptomatic with fever and mild hypotension 6 hours after receiving the first dose of aldesleukin. The patient had pretreatment pneumonia with RUL consolidation that was treated with oral antibiotics. The patient had mild symptoms at the time of receiving the second dose of AU-007 + aldesleukin and continued treatment. [Figure 42C] FIG. 1 presents a chart detailing drug-related adverse events (as of October 13, 2023) in all three arms (1A, 1B, and 1C).

[0095] [Figure 43A]Arm 1B AU-007+Proleukin® (aldesleukin): mild toxicity profile of drug-related adverse events (AEs). [Figure 43B] Arm 1B AU-007 + Proleukin® (aldesleukin): mild toxicity profile of drug-related adverse events (AEs). In Figure 43B, dMMR indicates mismatch repair deficiency.

[0096] Figures 44A, 44B, and 44C show AU-007 objective response: waterfall plots showing ongoing results for patients in the study suffering from different cancers as of October 13, 2023. [Figure 44A] FIG. 1 shows tumor status and subject enrollment within study, AU-007 monotherapy (Arm 1A): Best % change relative to baseline. [Figure 44B] Figure 1 shows AU-007 + Aldesleukin: Best % Change relative to baseline for all response-evaluable patients who received AU-007 + Aldesleukin. **Patient had a new brain lesion that was stabilized with radiation. [Figure 44C] Figure 1 shows AU-007 + aldesleukin for baseline immune-sensitive tumors (GI cancer excluded): Best % change. This includes all response-evaluable patients with non-GI cancer who received AU-007 + aldesleukin.

[0097] [Figure 45A] AU-007 Spider Plot: Percent (%) Tumor Change Over Time. [Figure 45B] AU-007 + Aldesleukin (IL-2): Percent (%) Tumor Change over Time. Figure 45B includes all response-evaluable patients who received AU-007 + Aldesleukin (as of October 13, 2023).

[0098] [Figure 46A]FIG. 1 shows tumor assessment by computed tomography scans (baseline and week 8 scans) from a melanoma patient whose cancer did not respond to checkpoint inhibitors anti-PD-1 and / or CTLA4. [Figure 46B] FIG. 1 shows tumor assessment by computed tomography scans (baseline and week 8 scans) from a melanoma patient whose cancer did not respond to checkpoint inhibitors anti-PD-1 and / or CTLA4. [Figure 46C] FIG. 1 shows tumor assessment by computed tomography scans (baseline and week 8 scans) from a melanoma patient whose cancer did not respond to checkpoint inhibitors anti-PD-1 and / or CTLA4.

[0099] [Figure 47] Figure 1 shows a graph of AU-007 concentrations over time. The left panel is an expansion of the first 60 hours showing Tmax and C-max. The right panel represents the data set as of June 2023. Note that this data is currently being acquired, so not all cohorts have complete data. Overall, the data demonstrate that AU007 exhibits typical IgG1 therapeutic properties.

[0100] Figures 48A and 48B show AU-007 pharmacodynamic data demonstrating that AU-007 continues to reduce peripheral blood circulating Tregs (measured by flow cytometry) over time (days) following administration of AU-007. Percent change in absolute numbers of circulating regulatory T cells. Regulatory T cells were defined as CD3+CD4+CD25+CD127lo in CD45+ cells. Consistent with a mechanism of action that inhibits IL-2 interaction with its trimeric receptor, regulatory T cells were reduced in the peripheral circulation. This was observed in both the monotherapy arm and the arm that also included Proleukin and was consistent across patients. [Figure 48A]FIG. 49 shows data by individual patient (receiving at least one dose of AU-007+ / -Proleukin - see legend in FIG. 49). [Figure 48B] Figure 1 shows data by treatment group (AU-007 alone). Values ​​represent change from baseline in absolute (abs) cell counts.

[0101] [Figure 49]

[0023] Figure 1 shows the mean percent change / day in peripheral blood Tregs for the cohort that received at least one dose of Proleukin® (aldesleukin) with 4.5 mg / kg AU-007. Values ​​represent the change from baseline in absolute (abs) cell counts. Data points continue to be collected.

[0102] [Figure 50A] FIG. 1 shows the changes in absolute peripheral blood CD8 cells over time (days) after administration of AU-007. [Figure 50B] Figure 50 shows the change in absolute peripheral blood CD8 cells over time (days) following administration of AU-007. Figure 50A shows data by individual patient, and Figure 50B shows data by treatment group. Values ​​represent the change from baseline in absolute (abs) cell counts. Data collection is ongoing.

[0103] [Figure 51]

[0023] Figure 1 shows the mean percent change / day in peripheral blood CD8 for the cohort receiving Proleukin® (aldesleukin) along with AU-007. Values ​​represent the change from baseline in absolute (abs) cell counts. Data collection is ongoing.

[0104] [Figure 52A] FIG. 1 shows changes in absolute peripheral blood NK cells over time (days) after administration of AU-007. [Figure 52B]Figure 52 shows the change in absolute peripheral blood NK cells over time (days) following administration of AU-007. Figure 52A shows data by individual patient, and Figure 52B shows data by treatment group. Values ​​represent the change from baseline in absolute (abs) cell counts. Data collection is ongoing.

[0105] [Figure 53] Figure 1 shows the change in peripheral blood NK cells for the Proleukin® (aldesleukin)-treated cohort over time (days) following administration of AU-007. Values ​​represent the change from baseline in absolute (abs) cell counts. Data as of September 2023

[0106] [Figure 54A] FIG. 1 shows absolute eosinophil counts over time (days) after administration of AU-007. [Figure 54B] Figures 54A and 54B show absolute eosinophil counts over time (days) following administration of AU-007. Figure 54A shows individual peripheral blood eosinophil counts in the AU-007 alone cohort. Figure 54B shows individual peripheral blood eosinophil counts in the AU-007 + Proleukin cohort. Figures 54A and 54B show the change in circulating eosinophil counts over time. Figure 54A shows the cohort receiving AU-007 monotherapy alone, and Figure 54B shows the cohort receiving AU-007 with at least one dose of Proleukin. All but one patient in the AU-007 monotherapy and AU-007 + Proleukin arms showed no decline or change in circulating eosinophil levels. A patient in the 9 mg / kg cohort had severe seasonal allergies requiring treatment during AU-007 treatment, consistent with a history of treatment for seasonal allergies. The increase in eosinophils was due to an allergic reaction. All patients receiving AU-007 plus Proleukin showed stable or decreased circulating eosinophils, consistent with AU-007's mechanism of action, which is to prevent IL-2 from interacting with the IL-2 trimeric receptor on eosinophils. Values ​​represent change from baseline in absolute (abs) cell counts. Data collection is ongoing.

[0107] [Figure 55A] FIG. 1 shows the CD8:Treg ratio over time (days) in the periphery after administration of AU-007. [Figure 55B] FIG. 1 shows the CD8:Treg ratio over time (days) in the periphery after administration of AU-007. [Figure 55C] Figure 55 shows the CD8:Treg ratio over time (days) in the periphery after administration of AU-007. Figure 55A shows all available data for each individual patient (see Figure 55C for legend). Figure 55B presents data by dosing group. Figure 55C shows data for each individual patient who also received Proleukin® (aldesleukin). Consistent with the observations made with Treg and CD8+ T cell changes, a trend toward an increased CD8+ / Treg ratio with monotherapy is observed. In the presence of Proleukin, an increase in the CD8+ / Treg ratio was observed, particularly with higher doses of Proleukin. Consistent with the mechanism of action, there was a trend toward a higher CD8+ / Treg ratio with higher doses (of low-dose IL-2) and longer exposure, and no drug-related toxicity was observed. Increasing the dose of Proleukin is expected to further enhance peripheral responses. Values ​​represent the change from baseline in absolute (abs) cell counts. Data collection is ongoing.

[0108] [Figure 56A] FIG. 1 shows the fold change in IFN-γ expression in patients administered AU-007+ / -Proleukin. [Figure 56B]Figure 56A shows the fold change in IFN-γ expression in patients receiving AU-007 + / - Proleukin. Heatmap of the change from baseline in circulating levels of interferon gamma (IFN-γ). Light green represents a 20% to 2-fold change, medium green represents a 2- to 5-fold change, and dark green represents a >5-fold change. These preliminary results demonstrate that the longer patients receive monotherapy (Figure 56A), the more likely they are to have an increase in circulating IFN-γ. This is consistent with observations in circulating cell populations, particularly Treg and NK cells. The addition of low-dose IL-2 in the presence of AU-007 (Figure 56B) consistently increases IFN-γ in the peripheral circulation.

[0109] [Figure 57] FIG. 1 shows the hPBMC proliferation assay protocol.

[0110] [Figure 58A] FIG. 1 shows the fold change of CD4+ Tregs from CD4+ cells over time in vitro in the presence of control antibody (black), AU-007 (red), or CD25-IL-2 conjugate (green). [Figure 58B] Figure 58 shows the fold change in CD4+ Tregs from CD4+ cells over time in vitro in the presence of control antibody (black), AU-007 (red), or CD25-IL-2 conjugate (green). Figure 58A shows the results for low dose administration (10 pM), and Figure 58B shows the results for high dose administration (1 nM). The data demonstrate that both IL-2 alone or non-alpha CD25-conjugated IL-2 expand Treg populations, but in the presence of AU-007, there is no Treg expansion. Because non-alpha IL-2 does not bind to the trimeric receptor on Tregs, Treg population expansion is a direct result of IL-2 produced by T cells (endogenous IL-2). AU-007 not only inhibits exogenous IL-2, but also inhibits endogenous IL-2.

[0111] [Figure 59A]FIG. 1 shows the fold change of CD4+ Tregs from CD4+ cells over time in vitro in the presence of control antibody (blue) or AU-007 (green) in the absence of IL-2. [Figure 59B] Figure 59 shows the fold change in CD4+ Tregs from CD4+ cells over time in vitro in the presence of control antibody (blue) or AU-007 (green) in the absence of IL-2. Control wells from low-dose (Figure 59A) and high-dose (Figure 59B) IL-2 experiments in which no IL-2 was added. Graphs were normalized to no-treatment (i.e., media alone) wells. Results demonstrate that AU-007 inhibits the ability of Tregs to use low levels of endogenous IL-2.

[0112] [Figure 60A] FIG. 1 shows the fold change of CD4+ regulatory T cells-(CD3+CD4+CD127-CD25+FoxP3+) over time in vitro in the presence of control antibody (black), AU-007 (red), or CD25-IL-2 conjugate (green). [Figure 60B] Figure 60B shows the fold change of NK cells over time in vitro in the presence of control antibody (black), AU-007 (red), or CD25-IL-2 conjugate (green). (IL-2 and CD25-IL-2 conjugate were at 1 nM, and isotype control antibody and AU-007 were at 1 uM; legend to Figure 60B).

[0113] [Figure 61A] FIG. 1 shows fold change over time using mean fluorescence intensity (MFI) to show changes in a suppressive marker of Tregs (CD25). [Figure 61B] FIG. 1 shows fold change over time using mean fluorescence intensity (MFI) to show changes in a suppressive marker of Tregs (FoxP3). [Figure 61C] Figure 1 shows fold change over time using mean fluorescence intensity (MFI) to show changes in NK cell activation marker (CD56). Observations were taken 3-6 days after treatment.

[0114] [Figure 62A] Figure 1 shows the ratio of ΔCD8+ Teff:ΔTreg in the presence of low dose IL-2 (10 pM). [Figure 62B] Figure 1 shows the ratio of ΔCD8+ Teff:ΔTreg in the presence of high dose IL-2 (1 nM).

[0115] [Figure 63] FIG. 1 shows the experimental design scheme for assaying restimulation-induced cell death (RICD).

[0116] [Figure 64] FIG. 1 shows a graph showing percent (%) live lymphocytes over time (days) following administration of 1 nM IL-2 (black) or 1 nM IL-2 and 10 μM AU-007 (green).

[0117] [Figure 65] FIG. 1 shows the experimental design scheme for assaying tetanus toxoid (TT).

[0118] [Figure 66A] FIG. 1 graphically depicts the results of a TT assay at a low dose of TT (0.9 μg / ml TT). [Figure 66B] FIG. 1 graphically depicts the results of a TT assay at a high dose of TT (9.0 μg / ml of TT). [Figure 66C] FIG. 1 shows the fold change of Treg (CD25+) over time.

[0119] [Figure 67A] FIG. 1 is a graphical representation of restimulation-induced cell death (RICD) of CD8+ cells in cultures stimulated with the protein antigen tetanus toxoid (high dose - 9.0 μg / ml). [Figure 67B] FIG. 1 is a graphical representation of restimulation-induced cell death (RICD) of CD8+ cells in cultures stimulated with the protein antigen tetanus toxoid (low dose—0.9 μg / ml).

[0120] [Figure 68] FIG. 1 shows toxicokinetics (TK) from acute toxicity studies in cynomolgus monkeys (cyno).

[0121] [Figure 69A] FIG. 1 shows that AU-007 reduces CD4+CD25+ cells compared to placebo in a cynomolgus monkey study. [Figure 69B] FIG. 1 shows that AU-007 reduces CD4+CD25+ cells compared to placebo in a cynomolgus monkey study. [Figure 69C] Figure 69 shows that AU-007 reduces CD4+CD25+ cells compared to placebo in a cynomolgus monkey study. Figure 69A - PBS (n=12) vs. 5 mg / kg (n=8); Figure 69B - PBS (n=12) vs. 25 mg / kg (n=8); and Figure 69C - PBS (n=12) vs. 100 mg / kg (n=12). PBS values ​​are plotted in black on each graph. Points represent pre-dose values ​​on each dosing day. Means are + / - SEM. Each plot represents the percentage of CD25+ cells in the CD4+ population per cohort.

[0122] [Figure 70A] FIG. 1 shows that AU-007 increases peripheral NK cells compared to baseline in a cynomolgus monkey study. [Figure 70B] FIG. 1 shows that AU-007 increases peripheral NK cells compared to baseline in a cynomolgus monkey study. [Figure 70C] Figures 70A-70C show that AU-007 increases peripheral NK cells compared to baseline in a cynomolgus monkey study. Figure 70A - PBS (n=12) vs. 5 mg / kg (n=8); Figure 70B - PBS (n=12) vs. 25 mg / kg (n=8); and Figure 70C - PBS (n=12) vs. 100 mg / kg (n=12). PBS values ​​are plotted in black in each graph. Points represent pre-dose values ​​on each dosing day. Means are + / - SEM.

[0123] [Figure 71] FIG. 1 shows a dose-dependent increase in IL-2 in cynomolgus monkeys administered AU-007 compared to saline (PBS) controls.

[0124] [Figure 72] FIG. 1 shows simulated human pharmacokinetic (PK) profiles following a Q2W dosing regimen.

[0125] [Figure 73] FIG. 1 shows simulated steady-state human PK profiles based on a Q2W dosing regimen.

[0126] [Figure 74]

[0023] Figure 1 shows a table presenting details of human PK modeling of Q2W AU-007 dosing and IL-2 coverage. Reference superscripts - a: amount of IL-2 (IU / cc) that could bind to AU-007 based on linear 1:1 molecular stoichiometry; b: amount of HD IL-2 (600,000 IU / kg) based on an 80 kg patient (5 L blood volume) that could bind to AU-007 based on linear 1:1 molecular stoichiometry; c: between the 6th and 7th dose: 12-14 weeks; and d: amount of HD IL-2 (600,000 IU / kg) based on an 80 kg patient (5 L blood volume) that could bind to AU-007 considering 2 IL-2 molecules for 1 AU-007 molecule.

[0127] [Figure 75] FIG. 1 shows that administration of AU-007 in combination with subcutaneous administration of IL-2 (Proleukin® (aldesleukin)) is expected to deliver significantly more daily IL-2 to dimeric receptors on Teff and NK cells than can be achieved by competing products. DETAILED DESCRIPTION OF THE INVENTION

[0128] The present disclosure provides engineered anti-human IL-2 antibodies that bind human IL-2 with high affinity (e.g., 12.7 pM to 48 pM) to a predefined binding epitope. These antibodies bind IL-2 in a manner that completely prevents CD25 binding, while avoiding IL-2 binding to CD122, thereby modulating the immune response to immunostimulation by directly activating and expanding effector cells without interacting with CD25-expressing cells (e.g., regulatory T cells, short-lived cytotoxic T cells, pulmonary endothelial cells, and vascular endothelial cells). Thus, antibody / IL-2 complexes will drive robust immune responses against distinct viral loads or tumors by expanding and activating effector cells such as NK cells, central memory T cells, and virus- or tumor-specific T cells, while inhibiting IL-2 activation-induced cell death of short-lived CD25+ cytotoxic T cells, which are important for viral / tumor clearance. Antibody / IL-2 complexes will also reduce immunosuppression caused by the regulatory arm of the immune system. Furthermore, the antibody / IL-2 complexes will prevent undesired interactions of IL-2 with vascular and pulmonary CD25-expressing cells, thereby preventing the severe syndromes of IL-2-induced vascular leakage and IL-2-induced pulmonary edema frequently seen in models of viral pulmonary infection. In some embodiments, the activity of the modified anti-IL-2 antibodies described herein depends on the predefined epitopes to which they are designed to bind.

[0129] In some embodiments, the IL-2 antibodies disclosed herein block IL-2 binding to CD25. In some embodiments, the IL-2 antibodies disclosed herein bind to IL-2 and prevent newly secreted endogenous IL-2 from binding to Tregs, effectively blocking the negative feedback loop of IL-2 on Tregs. In some embodiments, the IL-2 antibodies disclosed herein prevent Treg proliferation. In some embodiments, the IL-2 antibodies disclosed herein block IL-2 binding to vascular endothelium. In some embodiments, the IL-2 antibodies disclosed herein block IL-2 binding to pulmonary endothelium. In some embodiments, the IL-2 antibodies disclosed herein block IL-2 binding to vascular and pulmonary endothelium.

[0130] Those skilled in the art will understand that in certain embodiments, the term "anti-IL-2 antibody" as used herein is interchangeable with the term "anti-human IL-2 antibody," all of which have the same qualities and meaning. Similarly, as used throughout, in certain embodiments, the term "IL-2" is interchangeable with the term "human IL-2," all of which have the same qualities and meaning.

[0131] In some embodiments, the anti-human IL-2 antibodies described herein inhibit the binding of IL-2 to the IL-2 receptor alpha (IL-2Rα, i.e., CD25) subunit, and thus inhibit binding to the trimeric IL-2Rαβγ receptor. In certain embodiments, anti-IL-2 antibodies that inhibit the binding of IL-2 to the trimeric IL-2 receptor (IL-2Rαβγ) do not inhibit the binding of IL-2 to the dimeric IL-2 receptor (IL-2Rβγ).

[0132] Figure 2 shows a schematic diagram of anti-IL-2 antibody-directed immunotherapy. Targeting IL-2 to different cell populations can be used to modulate immune responses toward immunosuppression or immune activation. The anti-human IL-2 antibodies disclosed herein are designed to bind with high affinity to the IL-2 epitope and block IL-2 binding to CD25. As a result, IL-2 is prevented from binding to short-lived CD8+ cytotoxic T cells or regulatory T cells that express high levels of CD25, and is redirected to preferentially bind to effector T cells, stimulating an enhanced immune response and improving viral or bacterial clearance. Furthermore, IL-2 binding to CD25-expressing endothelial cells is also blocked, which would prevent IL-2-induced pulmonary edema and vascular leakage.

[0133] In one embodiment, the present disclosure provides a method of treating a disease (e.g., a viral infection, a bacterial infection, or cancer) or condition (e.g., an undesirable condition caused by IL-2, such as, but not limited to, pulmonary edema) with an anti-IL-2 antibody designed to enhance T cell immune responses and prevent the severe edematous symptoms of IL-2-induced acute pneumonia. The anti-IL-2 antibody will specifically bind human IL-2 with high affinity at a predefined epitope that blocks IL-2 binding to the alpha chain of the IL-2 receptor (CD25), while avoiding binding to the major signaling beta and gamma chain complex of the receptor (CD122 / CD132). As a result, in the presence of such an antibody, IL-2 will be directed to immune cells responsible for viral / tumor clearance and away from cells that slow the immune response or cause edema. The formation of this IL-2 / antibody immune complex directs IL-2 to bind and activate exclusively naive and memory T lymphocytes, NK cells, and natural killer T lymphocytes, while preventing the activation of regulatory T cells and the apoptosis of short-lived CD25+ cytotoxic T effector cells. Overall, the end result is an effective immune response, e.g., viral or tumor clearance. Furthermore, this treatment would prevent toxicity caused by IL-2 binding to endothelial CD25-expressing cells. Thus, in one embodiment, targeting IL-2 with the anti-IL-2 antibodies disclosed herein would be an effective treatment for respiratory diseases caused by viral or bacterial infections. In another embodiment, treatment with the anti-IL-2 antibodies disclosed herein would be effective in preventing toxicity caused by IL-2 binding to endothelial CD25-expressing cells, e.g., pulmonary edema or IL-2-induced vascular leakage. More importantly, enhancing IL-2 immunostimulation toward general immune activation and proliferation of immune effector cells independent of specific pathogens (e.g., viral antigens) may be an effective strategy against future viral or bacterial pandemics caused by unknown pathogens (Figures 3A and 3B).

[0134] In one embodiment, the methods disclosed herein may be useful against infections caused by SARS-CoV-2. SARS-CoV-2 binds to angiotensin-converting enzyme 2 in lung cells, allowing the virus to enter and replicate. The immune response to pulmonary viral infections consists of both the innate and adaptive arms of the immune system. As with many respiratory viruses, clearance of SARS-CoV-2 from the lungs is expected to depend on the T cell immune response. The cytokine IL-2 is important for T cell proliferation and plays a key role in the immune response to viruses. However, in addition to its stimulatory role, IL-2 also induces several adverse side effects, such as pulmonary edema and vascular leak syndrome, through its binding to endothelium expressing the CD25 receptor.

[0135] Figures 3A and 3B show a schematic diagram of the progression of COVID-19 infection and potential anti-IL-2 therapy as an adjuvant intervention. Figure 3A shows that SARS-CoV-2 during infection causes nonsevere symptoms and induces a protective immune response after the incubation period. Successful elimination of the infection depends on the health of the infected individual. Individuals with an insufficient immune response to the virus have difficulty clearing it, while those with an overly strong immune response may develop pulmonary edema and other cytokine-mediated adverse effects. Therefore, strategies to enhance the immune response and prevent pulmonary edema are desirable. While high concentrations of IL-2 may be beneficial for viral clearance, especially in the early stages, high levels of IL-2 may lead to IL-2-induced pulmonary edema and vascular leakage through interactions between IL-2 and CD25-expressing endothelial cells. Figure 3B shows that anti-human IL-2 antibodies designed to bind to and block the CD25 / IL-2 interaction are predicted to enhance immune effector cell proliferation, improve viral clearance, and reduce the negative effects of IL-2 binding to CD25 expressed on endothelial cells, thereby preventing IL-2-induced pulmonary edema and vascular leakage.

[0136] Figure 1 shows a schematic of the mechanism of action of IL-2 and its dual role in regulating immune responses. The left panel shows that IL-2 consists of three epitope binding sites (α, β, γ) that interact with different forms of IL2-R (CD25, CD122, and CD132) with different affinities. The right panel shows that different IL-2R complexes are expressed on different T cell populations, and their different affinities for IL2 allow for immunosuppression under conditions of low local concentrations of IL-2 and immunostimulation when local concentrations of IL-2 are increased.

[0137] In the following detailed description, numerous specific details are set forth to provide a thorough understanding of the antibodies disclosed herein. However, it will be understood by those skilled in the art that the preparation and use of the antibodies disclosed herein may, in certain instances, be carried out without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the disclosure provided herein.

[0138] Throughout this application, various references or publications are cited. The disclosures of these references or publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

[0139] As used herein, the term "antibody" may be used interchangeably with the term "immunoglobulin," all of which have the same qualities and meaning. An antibody-binding domain or antigen-binding site may be a fragment of an antibody or a recombinant product of one or more fragments of an antibody, which fragments are involved in specific binding to a target antigen. "Specific binding" means that the binding is selective for the antigen of interest and can be distinguished from undesired or nonspecific interactions. For example, an equilibrium dissociation constant of ≦10 -5 , 10 -6 , or 10 -7 An antibody is said to specifically bind to an IL-2 epitope when M is . In some embodiments, the equilibrium dissociation constant is ≦10 -8 M or 10 -9In some further embodiments, the equilibrium dissociation constant may be ≦10 -10 M, 10 -11 M or 10 -12 In some embodiments, the equilibrium dissociation constant can be ≦10 -5 M~10 -12 M.

[0140] As used herein, the term "antibody" includes one or more antibody fragments that retain binding specificity, including, but not limited to, IgG, heavy chain variable region (VH), light chain variable region (VL), Fab fragments, F(ab')2 fragments, scFv fragments, Fv fragments, nanobodies, minibodies, diabodies, triabodies, tetrabodies, and single domain antibodies (see, e.g., Hudson and Souriau, Nature Med. 9:129-134 (2003)). These terms also encompass humanized antibodies, primatized antibodies, and chimeric antibodies, as generally understood in the art.

[0141] As used herein, the term "heavy chain variable region" may be used interchangeably with the term "VH domain" or "VH," all of which have the same meaning and qualities. As used herein, the term "light chain variable region" may be used interchangeably with the term "VL domain" or "VL," all of which have the same meaning and qualities. Those skilled in the art will recognize that a "heavy chain variable region" or "VH" in the context of an antibody encompasses a fragment of a heavy chain comprising three complementarity-determining regions (CDRs) sandwiched between adjacent stretches known as framework regions. Framework regions are more highly conserved than the CDRs and form a scaffold to support the CDRs. Similarly, those skilled in the art will recognize that a "light chain variable region" or "VL" in the context of an antibody encompasses a fragment of a light chain comprising three CDRs sandwiched between framework regions.

[0142] As used herein, the term "complementarity-determining region" or "CDR" refers to the hypervariable region of a heavy or light chain variable region. Each heavy or light chain polypeptide has three CDRs, designated "CDR1," "CDR2," and "CDR3," proceeding from the N-terminus. Crystallographic analysis of several antigen-antibody complexes has demonstrated that the amino acid residues of the CDRs form extensive contacts with the bound antigen, with the most extensive antigen contact being with the heavy chain CDR3. Thus, the CDR regions are primarily responsible for the specificity of the antigen-binding site. In one embodiment, the antigen-binding site comprises six CDRs, including CDRs from each of the heavy and light chain variable regions.

[0143] As used herein, the term "framework region" or "FR" refers to the four adjacent amino acid sequences surrounding the CDRs of a heavy or light chain variable region. While some FR residues may contact the bound antigen, FR residues primarily play a role in folding the variable region into the antigen-binding site. In some embodiments, FR residues responsible for folding the variable region include residues directly adjacent to the CDRs. Within FRs, certain amino acid residues and certain structural features are highly conserved. In this regard, all variable region sequences contain an internal disulfide loop of approximately 90 amino acid residues. When the variable region folds into the antigen-binding site, the CDRs are displayed as protruding loop motifs that form the antigen-binding surface. Regardless of the exact CDR amino acid sequence, it is generally recognized that there are conserved structural regions of FRs that affect the folding of the CDR loops into certain "canonical" structures. Furthermore, certain FR residues are known to participate in noncovalent interdomain contacts that stabilize the interaction between the heavy and light chains of an antibody.

[0144] Wu and Kabat (Tai Te Wu, Elvin A. Kabat. An analysis of the sequences of the variable regions of Benedict Jones proteins and myeloma light chains and their implications for antibody complementarity. Journal of Experimental Medicine, 132, 2, 8 (1970); Kabat EA, Wu TT, Bilofsky H, Reid-Miller M, Perry H. Sequence of proteins of immunological interest. Bethesda: National Institutes of Health; 1983, 323 (1983)) pioneered the alignment of antibody peptide sequences, and their contributions in this regard were several-fold: First, through studies of sequence similarity between variable domains, they identified corresponding residues that are more or less homologous across all antibodies of all vertebrate species because they adopt similar three-dimensional structures, play similar functional roles, interact similarly with neighboring residues, and exist in similar chemical environments. Second, they devised a peptide sequence numbering system that assigns the same position number to homologous immunoglobulin residues. Those skilled in the art can unambiguously assign what is now commonly called the Kabat numbering to any variable domain sequence without relying on experimental data beyond the sequence itself. Third, Kabat and Wu calculated the variability for each Kabat-numbered sequence position, which means finding a small or large number of possible amino acids when variable domain sequences are aligned. They identified three highly variable flanking regions embedded within four less variable flanking regions. Kabat and Wu formally distinguished the residues that make up these variable regions and designated them "complementarity-determining regions" (CDRs), referring to the chemical complementarity between antibody and antigen. The role in the three-dimensional folding of the variable domain, rather than in antigen recognition, is attributed to the remaining less variable regions, referred to herein as "framework regions."Fourth, Kabat and Wu established a public database of antibody peptide and nucleic acid sequences that continues to be maintained and is well known to those skilled in the art.

[0145] Chothia and coworkers (Cyrus Chothia, Arthur M. Lesk. Canonical structures for the hypervariable regions of immunoglobulins. Journal of Molecular Biology, 196, 4, 8 (1987)) found that certain subportions within the Kabat CDRs adopt nearly identical peptide backbone conformations despite great diversity at the amino acid sequence level. These subportions are designated L1, L2, and L3 or H1, H2, and H3, where "L" and "H" indicate the light chain and heavy chain regions, respectively. These regions may be referred to as Chothia CDRs, whose boundaries overlap with the Kabat CDRs.

[0146] More recent studies have shown that virtually all antibody-binding residues fall within regions of structural consensus (Kunik, V. et al., PloS Computational Biology 8(2):el002388 (February 2012)). In some embodiments, these regions are referred to as antibody-binding regions. It has also been shown that these regions can be identified from antibody sequences. For this purpose, "Paratome," a structural approach to identifying structural consensus in antibodies, was used (Ofran, Y. et al., J. Immunol. 757:6230-6235 (2008)). While residues identified by Paratome cover virtually the entire antibody-binding site, CDRs (as identified by commonly used CDR identification tools) miss a significant portion of them. Antibody-binding residues identified by Paratome but not by any of the common CDR identification methods are referred to as Paratome-unique residues. Similarly, antibody-binding residues identified by any of the common CDR identification methods but not by Paratome are referred to as CDR-unique residues. Paratome-specific residues make significant energetic contributions to antibody-antigen interactions, whereas CDR-specific residues make significantly smaller contributions. These results allow for better definition of the antigen-binding site.

[0147] IMGT® is the International ImMunoGeneTics Information System® (see Nucleic Acids Res. 2015 Jan;43(Database issue):D413-22. doi:10.1093 / nar / gku1056. Epub 2014 Nov 5 Free article. PMID:25378316 LIGM:441 and Dev Comp Immunol. 2003 Jan;27(1):55-77). IMGT is a unique numbering system for immunoglobulin and T-cell receptor variable domains and Ig superfamily V-like domains (Lefranc et al., Dev Comp Immunol. 27:55-77(2003)). IMGT® provides a uniform numbering system for IG and TcR variable domain sequences based on aligning five or more IG and TcR variable region sequences, taking into account and combining Kabat definitions of FRs and CDRs, structural data, and Chothia characterization of hypervariable loops. IMGT is considered to be well known in the art as a universal numbering scheme for antibodies.

[0148] In some embodiments, the identification of potential variant amino acid positions in the VH and VL domains uses the IMGT analysis system. In some embodiments, the identification of potential variant amino acid positions in the VH and VL domains uses the Paratome analysis system. In some embodiments, the identification of potential variant amino acid positions in the VH and VL domains uses the Kabat analysis system. In some embodiments, the identification of potential variant amino acid positions in the VH and VL domains uses the Clothia analysis system.

[0149] In describing the mutated amino acid positions present in the VH and VL domains, in some embodiments, IMGT numbering is used. In describing the mutated amino acid positions present in the VH and VL domains, in some embodiments, Paratome numbering is used. In describing the mutated amino acid positions present in the VH and VL domains, in some embodiments, Kabat numbering is used. In describing the mutated amino acid positions present in the VH and VL domains, in some embodiments, Clothia numbering is used.

[0150] Antigen-binding sequences are traditionally located within the heavy and light chain variable regions of an antibody. These heavy and light chain variable regions can, in certain cases, be engineered to create new binding sites, for example, to create antibodies or fragments thereof that bind to different antigens or different epitopes of the same antigen. In some embodiments, as described herein, a new binding site for a second antigen will be created by engineering the heavy chain variable region sequence or the light chain variable region sequence, or both.

[0151] Antibodies may exist in various forms or have various domains, including, but not limited to, complementarity determining regions (CDRs), variable regions (Fv), VH domains, VL domains, single chain variable regions (scFv), and Fab fragments.

[0152] Those skilled in the art will appreciate that an scFv is a fusion polypeptide comprising the variable heavy (VH) and variable light (VL) regions of an immunoglobulin connected by a short linker peptide, which linker may have, for example, from 10 to about 25 amino acids.

[0153] Those skilled in the art will also understand that the term "Fab" in reference to an antibody generally encompasses the portion of an antibody consisting of a single light chain (both variable and constant regions) linked by disulfide bonds to the variable region and first constant region of a single heavy chain, and that F(ab')2 comprises a fragment of the heavy chain containing the VH domain and a fragment of the light chain containing the VL domain.

[0154] In some embodiments, antibodies include whole antibody molecules, including monoclonal and polyclonal antibodies. In some embodiments, antibodies include one or more antibody fragments that retain binding specificity, including, but not limited to, variable heavy chain (VH) fragments, variable light chain (VL) fragments, Fab fragments, F(ab')2 fragments, scFv fragments, Fv fragments, minibodies, diabodies, triabodies, and tetrabodies.

[0155] Modified anti-IL-2 antibody In one embodiment, the present disclosure provides modified anti-IL-2 antibodies resulting from introducing amino acid mutations into a parent anti-IL-2 antibody. In one embodiment, one or more of the amino acid mutations are introduced in the CDR region. In another embodiment, one or more amino acid mutations are introduced within the framework (FR) region. In yet another embodiment, amino acid mutations are introduced in both the CDR and framework (FR) regions. One of skill in the art would readily use a variety of standard techniques known in the art to introduce amino acid mutations into an anti-IL-2 antibody and then test the resulting modified antibody for any changes in binding to IL-2. While standard techniques may be used, the resulting binding pattern of the newly generated antibody is not predictable and must be analyzed to determine functionality.

[0156] In certain embodiments, the present disclosure provides polypeptides comprising a VH domain and a VL domain that can be dimerized under appropriate conditions. For example, the VH domain and the VL domain can be combined in an appropriate buffer and dimerized by appropriate interactions, such as hydrophobic interactions. In another embodiment, the VH domain and the VL domain can be combined in an appropriate buffer containing an enzyme and / or cofactor that can promote dimerization of the VH domain and the VL domain. In another embodiment, the VH domain and the VL domain can be combined in an appropriate vehicle that allows them to react with each other in the presence of appropriate reagents and / or catalysts.

[0157] In certain embodiments, the VH and VL domains may be comprised within a longer polypeptide sequence, including, but not limited to, a constant region, hinge region, linker region, Fc region, or disulfide bond region, or any combination thereof. A constant domain is an immunoglobulin fold unit of the constant portion of an immunoglobulin molecule, also referred to as a domain of the constant region (e.g., CH1, CH2, CH3, CH4, Ck, Cl). In some embodiments, the longer polypeptide may include multiple copies of one or both of the VH and VL domains generated according to the methods disclosed herein, for example, when the polypeptides generated herein are used to form diabodies or triabodies.

[0158] In some embodiments, the Fc region comprises at least one mutation that reduces Fc-gamma binding, i.e., binding to Fcγ receptors (FcγRs). In some embodiments, the reduced binding is abrogated, i.e., undetectable binding to Fcγ receptors. In some embodiments, the reduced binding reduces binding affinity to Fcγ receptors. In some embodiments, the reduced binding reduces the on-rate of binding to Fcγ receptors. In some embodiments, the reduced binding reduces the off-rate of binding to Fcγ receptors. In some embodiments, the mutation that reduces Fc-gamma binding comprises L234A, L235A mutations, also known as LALA mutations. In some embodiments, the mutation that reduces Fc-gamma binding comprises a P329G mutation in addition to the L234A, L235A mutations. In some embodiments, the antibodies described herein comprise a heavy chain comprising a mutation that reduces binding to Fcγ receptors.

[0159] In one embodiment, the disclosure provides an engineered (or modified) anti-IL-2 antibody comprising a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In one embodiment, the engineered antibody can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, or F(ab')2. The IgG can be of the IgG1, IgG2, IgG3, or IgG4 subclass. In another embodiment, the engineered antibody can be part of a minibody, diabody, or triabody antibody.

[0160] In one embodiment, the disclosure provides an engineered (or modified) anti-IL-2 antibody comprising a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In one embodiment, the engineered antibody can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, or F(ab')2. The IgG can be of the IgG1, IgG2, IgG3, or IgG4 subclass. In another embodiment, the engineered antibody can be part of a minibody, diabody, or triabody antibody.

[0161] In one embodiment, the present disclosure provides an engineered (or modified) anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27, or SEQ ID NOs: 36 and 37. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 10 and 11. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 12 and 13. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 14 and 15. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 16 and 17. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 18 and 19. In one embodiment, the engineered anti-IL-2 antibody comprises the sequence of SEQ ID NOs: 20 and 21. In one embodiment, the modified anti-IL-2 antibody comprises the sequences of SEQ ID NOs: 22 and 23. In one embodiment, the modified anti-IL-2 antibody comprises the sequences of SEQ ID NOs: 24 and 25. In one embodiment, the modified anti-IL-2 antibody comprises the sequences of SEQ ID NOs: 26 and 27. In one embodiment, the modified anti-IL-2 antibody comprises the sequences of SEQ ID NOs: 36 and 37.

[0162] In some embodiments, the isolated anti-IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (b) HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (c) HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; (d) HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; or (e) HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, LCDR2 comprises the amino acid sequence of DAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; It has the amino acid sequence:

[0163] In some embodiments, the isolated anti-IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are (a) HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, LCDR2 comprises the amino acid sequence of YAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; It has the amino acid sequence:

[0164] In some embodiments, the VH and VL have amino acid sequences in which VH comprises the amino acid sequence of SEQ ID NO: 10 and VL comprises the amino acid sequence of SEQ ID NO: 11; VH comprises the amino acid sequence of SEQ ID NO: 12 and VL comprises the amino acid sequence of SEQ ID NO: 13; VH comprises the amino acid sequence of SEQ ID NO: 14 and VL comprises the amino acid sequence of SEQ ID NO: 15; VH comprises the amino acid sequence of SEQ ID NO: 16 and VL comprises the amino acid sequence of SEQ ID NO: 17; VH comprises the amino acid sequence of SEQ ID NO: 18 and VL comprises the amino acid sequence of SEQ ID NO: 19; VH comprises the amino acid sequence of SEQ ID NO: 20 and VL comprises the amino acid sequence of SEQ ID NO: 21; VH comprises the amino acid sequence of SEQ ID NO: 22 and VL comprises the amino acid sequence of SEQ ID NO: 23; VH comprises the amino acid sequence of SEQ ID NO: 24 and VL comprises the amino acid sequence of SEQ ID NO: 25; VH comprises the amino acid sequence of SEQ ID NO: 26 and VL comprises the amino acid sequence of SEQ ID NO: 27; or VH comprises the amino acid sequence of SEQ ID NO: 36 and VL comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the VH and VL have amino acid sequences, where the VH comprises the amino acid sequence of SEQ ID NO:26 and the VL comprises the amino acid sequence of SEQ ID NO:27.

[0165] In some embodiments, the antibody comprises a heavy chain sequence and a light chain sequence, wherein the heavy chain sequence is set forth in SEQ ID NO: 68 and the light chain sequence is set forth in SEQ ID NO: 69; the heavy chain sequence is set forth in SEQ ID NO: 70 and the light chain sequence is set forth in SEQ ID NO: 71; or the heavy chain sequence is set forth in SEQ ID NO: 72 and the light chain sequence is set forth in SEQ ID NO: 73. In some embodiments, the antibody comprises a heavy chain sequence and a light chain sequence, wherein the heavy chain sequence is set forth in SEQ ID NO: 68 and the light chain sequence is set forth in SEQ ID NO: 69. In some embodiments, the antibody comprises a heavy chain sequence and a light chain sequence, wherein the heavy chain sequence is set forth in SEQ ID NO: 70 and the light chain sequence is set forth in SEQ ID NO: 71. In some embodiments, the antibody comprises a heavy chain sequence and a light chain sequence, wherein the heavy chain sequence is set forth in SEQ ID NO: 72 and the light chain sequence is set forth in SEQ ID NO: 73.

[0166] In one embodiment, the modified antibody can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, or F(ab')2. The IgG can be of the IgG1, IgG2, IgG3, or IgG4 subclass. In another embodiment, the modified antibody can be part of a minibody, diabody, or triabody antibody.

[0167] In one embodiment, the present disclosure also provides an isolated polynucleotide sequence encoding a heavy chain variable region of an anti-IL-2 antibody, wherein the heavy chain variable region comprises one of the amino acid sequences of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In another embodiment, the present disclosure also provides a vector comprising the above-described polynucleotide sequence. In view of the amino acid sequences disclosed herein, one skilled in the art would readily construct a vector or plasmid encoding the amino acid sequence. In another embodiment, the present disclosure also provides a host cell comprising the vector provided herein. Depending on the use and experimental conditions, one skilled in the art would readily use a host cell suitable for harboring and / or expressing the above-described polynucleotide sequence.

[0168] In one embodiment, the present disclosure also provides an isolated polynucleotide sequence encoding a light chain variable region of an anti-IL-2 antibody, wherein the light chain variable region comprises one of the amino acid sequences of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In another embodiment, the present disclosure also provides a vector comprising the above-described polynucleotide sequence. In view of the amino acid sequences disclosed herein, one skilled in the art would readily construct a vector or plasmid encoding the amino acid sequence. In another embodiment, the present disclosure also provides a host cell comprising the vector provided herein. Depending on the use and experimental conditions, one skilled in the art would readily use a host cell suitable for harboring and / or expressing the above-described polynucleotide sequence.

[0169] Given the heavy and light chain variable region sequences disclosed herein, one of skill in the art would readily use standard techniques known in the art to construct an anti-IL-2 scFv. In one embodiment, a polynucleotide sequence encoding such an anti-IL-2 scFv could have the sequence of one of SEQ ID NOS: 1-5 or one of SEQ ID NOS: 31-35.

[0170] In certain embodiments, the isolated polynucleotide sequence disclosed herein encoding the heavy chain variable region of an anti-IL-2 antibody comprises the VH amino acid sequence set forth in any of the amino acid sequences of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, a vector comprises the polynucleotide sequence of any of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, a host cell comprises a vector comprising the polynucleotide sequence of any of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36.

[0171] In certain embodiments, the isolated polynucleotide sequence disclosed herein encoding the light chain variable region of an anti-IL-2 antibody comprises the VL amino acid sequence set forth in any of the amino acid sequences of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, the vector comprises a polynucleotide sequence comprising the amino acid sequence of any of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, the host cell comprises a vector comprising a polynucleotide sequence encoding the amino acid sequence of any of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, the isolated polynucleotide sequence encodes an anti-IL-2 scFv, and the polynucleotide sequence is set forth in SEQ ID NOs: 1, 2, 3, 4, 5, 31, 32, 33, 34, or 35. In some embodiments, the vector comprises an isolated polynucleotide sequence encoding an anti-IL-2 scFv, wherein the polynucleotide sequence is set forth in SEQ ID NO: 1, 2, 3, 4, 5, 31, 32, 33, 34, or 35. In some embodiments, the host cell comprises a vector comprising an isolated polynucleotide sequence encoding an anti-IL-2 scFv, wherein the polynucleotide sequence is set forth in SEQ ID NO: 1, 2, 3, 4, 5, 31, 32, 33, 34, or 35.

[0172] In another embodiment, the present disclosure also provides an isolated anti-IL-2 antibody comprising a heavy chain variable region having complementarity-determining regions (CDRs) 1, 2, and 3. In one embodiment, CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38-40, respectively; 44-46, respectively; 50-52, respectively; 56-58, respectively; or 62-64, respectively. In one embodiment, the antibody can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab'), minibody, diabody, or triabody antibody. The IgG can be IgG1, IgG2, IgG3, or IgG4. In one embodiment, the present disclosure also encompasses a composition comprising the above-described antibody and a pharmaceutically acceptable carrier.

[0173] In another embodiment, the present disclosure also provides an isolated anti-IL-2 antibody comprising a light chain variable region having complementarity-determining regions (CDRs) 1, 2, and 3. In one embodiment, CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41-43, respectively; 47-49, respectively; 53-55, respectively; 59-61, respectively; or 65-67, respectively. In one embodiment, the antibody can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab'), minibody, diabody, or triabody antibody. The IgG can be IgG1, IgG2, IgG3, or IgG4. In one embodiment, the present disclosure also encompasses a composition comprising the above-described antibody and a pharmaceutically acceptable carrier.

[0174] In another embodiment, the present disclosure also provides an isolated anti-IL-2 antibody comprising a heavy chain variable region having complementarity-determining regions (CDRs) 1, 2, and 3, and a light chain variable region having CDR1, 2, and 3. In one embodiment, the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38-40, 44-46, 50-52, 56-58, or 62-64, respectively. In one embodiment, the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41-43, 47-49, 53-55, 59-61, or 65-67, respectively. In one embodiment, the antibody may be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab'), minibody, diabody, or triabody antibody. The IgG can be IgG1, IgG2, IgG3, or IgG4. In one embodiment, the present disclosure also encompasses a composition comprising the above-described antibody and a pharmaceutically acceptable carrier.

[0175] Pharmaceutical Composition In some embodiments, compositions for therapeutic use are disclosed herein. In some embodiments, the compositions described herein comprise an anti-IL-2 antibody disclosed herein and a pharmaceutically acceptable carrier.

[0176] As used herein, the terms "composition" and "pharmaceutical composition" may, in some embodiments, be used interchangeably, all with the same quality and meaning. Disclosed herein, in some embodiments, are pharmaceutical compositions for treating a condition or disease described herein.

[0177] In some embodiments, disclosed herein are pharmaceutical compositions for use in combination therapy.

[0178] In another embodiment, disclosed herein are compositions for use in treating a disease or condition in a subject. In some embodiments, the disease comprises a viral infection, a bacterial infection, or cancer. In some embodiments, the condition comprises an IL-2-induced condition. In some embodiments, the IL-2-induced condition comprises pulmonary edema or vascular leakage.

[0179] Administration The VH and / or VL polypeptides disclosed herein can be administered to a subject (e.g., a human or animal) alone or in combination with a carrier, i.e., a pharmaceutically acceptable carrier. Pharmaceutically acceptable means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to a subject without causing undesired biological effects or adversely interacting with any of the other components of the pharmaceutical composition in which it is contained. As is well known to those of skill in the art, the carrier will be selected to minimize any degradation of the polypeptides disclosed herein and to minimize any adverse side effects in the subject. Pharmaceutical compositions can be prepared by methodology well known in the pharmaceutical arts.

[0180] The above-described pharmaceutical compositions comprising the polypeptides disclosed herein can be administered in any suitable manner (e.g., to a mammal, cell, or tissue) depending on whether local or systemic treatment is desired. For example, the compositions can be administered topically (e.g., ophthalmically, vaginally, rectally, intranasally, transdermally, etc.), orally, by inhalation, or parenterally (including intravenous infusion or subcutaneous, intracavity, intraperitoneal, intradermal, or intramuscular injection). Local intranasal administration refers to delivery of the composition to the nose and nasal cavity through one or both nostrils. The composition can be delivered by a spray or droplet mechanism or through aerosolization. Delivery can also be directed to any region of the respiratory system (e.g., the lungs) via intubation. Alternatively, administration can be intratumoral, e.g., by topical or intravenous injection.

[0181] When the composition is administered parenterally, it is generally administered by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for suspension in liquid before injection, or emulsions. Furthermore, parenteral administration can involve the preparation of delayed or sustained release systems to maintain a constant dosage.

[0182] In some embodiments, a composition comprises an anti-IL-2 antibody comprising a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, a composition comprises an anti-IL-2 antibody comprising a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, a composition comprises an anti-IL-2 antibody comprising heavy and light chain variable regions having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27, or SEQ ID NOs: 36 and 37. In some embodiments, the composition comprises an anti-IL-2 antibody comprising a heavy chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 38-40, respectively; 44-46, respectively; 50-52, respectively; 56-58, respectively; or 62-64, respectively. In some embodiments, the composition comprises an anti-IL-2 antibody comprising a light chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 41-43, respectively; 47-49, respectively; 53-55, respectively; 59-61, respectively; or 65-67, respectively. In some embodiments, the composition comprises an anti-IL-2 antibody comprising a heavy chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; SEQ ID NOs: 44 to 46, respectively; SEQ ID NOs: 50 to 52, respectively; SEQ ID NOs: 56 to 58, respectively; or SEQ ID NOs: 62 to 64, respectively; and a light chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; SEQ ID NOs: 47 to 49, respectively; SEQ ID NOs: 53 to 55, respectively; SEQ ID NOs: 59 to 61, respectively; or SEQ ID NOs: 65 to 67, respectively.

[0183] In some embodiments, the composition comprises an anti-IL-2 antibody comprising any of clones BDG 17.014, BDG 17.023, BDG 17.038, BDG 17.043, BDG 17.053, BDG 17.054, BDG 17.066, BDG 17.067, and BDG 17.069.

[0184] In one embodiment, the methods disclosed herein involve administering a composition comprising an anti-IL-2 antibody disclosed herein as a monotherapy. In another embodiment, the methods disclosed herein involve administering a composition comprising an anti-IL-2 antibody disclosed herein in combination with a single dose of IL-2. In another embodiment, the methods disclosed herein involve administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and a composition comprising IL-2.

[0185] In some embodiments, when an anti-IL2 antibody and IL-2 are administered, they may be administered in the same composition. In some embodiments, when an anti-IL2 antibody and IL-2 are administered, they may be administered in separate compositions.

[0186] In some embodiments, IL-2 may be administered before, simultaneously with, or after administering an anti-IL2 antibody. In some embodiments, IL-2 administration is before administering an anti-IL2 antibody. In some embodiments, IL-2 administration is concurrent with administering an anti-IL2 antibody. In some embodiments, IL-2 administration follows administering an anti-IL2 antibody.

[0187] In some embodiments, the route of IL-2 administration is subcutaneous. In some embodiments, the subcutaneous administration of IL-2 is at a much lower dose and much less frequent than the approved regimen of intravenously administered aldesleukin. In some embodiments, the route of administration of anti-IL-2 is by intravenous injection. In some embodiments, in which a subject is administered both an anti-IL-2 antibody and IL-2, the route of administration of the anti-IL-2 antibody is by intravenous injection and the route of administration of the IL-2 is by subcutaneous injection.

[0188] In some embodiments, administering the anti-IL-2 antibody comprises monotherapy. In some embodiments, administering the anti-IL-2 antibody comprises including a loading dose of IL-2 in anti-IL-2 antibody monotherapy. In some embodiments, administering the anti-IL-2 antibody comprises combination therapy in which an anti-IL-2 antibody and IL-2 are administered to the subject at regular intervals.

[0189] In some embodiments, multiple doses of anti-IL2 antibody are administered over a given period of time. In some embodiments, doses of anti-IL2 antibody are administered weekly. In some embodiments, doses of anti-IL2 antibody are administered every other week (once every two weeks). In some embodiments, doses of anti-IL2 antibody are administered once every three weeks. In some embodiments, a single dose of IL-2 may be administered before, simultaneously with, or after the first dose of an anti-IL2 antibody described herein. In some embodiments, multiple doses of anti-IL2 antibody and IL-2 are administered over a given period of time. In some embodiments, doses of anti-IL2 antibody and IL-2 are administered weekly. In some embodiments, doses of anti-IL2 antibody and IL-2 are administered every other week (once every two weeks). In some embodiments, doses of anti-IL2 antibody and IL-2 are administered once every three weeks.

[0190] In some embodiments, the anti-IL-2 antibody is administered weekly, every other week, or every three weeks, and IL-2 is administered as a single dose once every week, every other week, or every three weeks, or the anti-IL-2 antibody and IL-2 are administered simultaneously. In some embodiments, the anti-IL-2 antibody is administered weekly, every other week, or every three weeks, and IL-2 is administered as a single dose once every week, every other week, or every three weeks, or the anti-IL-2 antibody and IL-2 are administered independently of each other, for example, but not limited to, the anti-IL-2 antibody is administered weekly, every other week, or every three weeks, and IL-2 is administered as a single dose, or the anti-IL-2 antibody is administered weekly, every other week, or every three weeks, and IL-2 is administered weekly, or the anti-IL-2 antibody is administered weekly, every other week, or every three weeks, and IL-2 is administered every three weeks.

[0191] In some embodiments, the therapeutic dose of anti-IL-2 is administered over a period of several months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for up to 3 months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for at least 3 months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for up to 6 months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for at least 6 months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for up to 9 months. In some embodiments, the therapeutic dose of anti-IL-2 is administered for at least 9 months.

[0192] In some embodiments, the therapeutic dose of anti-IL-2 is administered for a period of up to one year. In some embodiments, the therapeutic dose of anti-IL-2 is administered for a period of at least one year.

[0193] In some embodiments, an anti-IL-2 antibody disclosed herein is administered in combination with IL-2, and the dose of IL-2 is considered a low dose of IL-2. In some embodiments, IL-2 is administered as a single dose (loading dose). In some embodiments, IL-2 is administered over the same period as the anti-IL-2 antibody. In some embodiments, IL-2 is administered as multiple doses before, simultaneously with, or after administration of the anti-IL-2 antibody.

[0194] In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is about 0.5 mg / kg to 12 mg / kg, or about 0.5 mg / kg, 1.0 mg / kg, 1.5 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 3.0 mg / kg, 3.5 mg / kg, 4.0 mg / kg, 4.5 mg / kg, 5.0 mg / kg, 5.5 mg / kg, 6.0 mg / kg, 6.5 mg / kg, 7.0 mg / kg, 7.5 mg / kg, 8.0 mg / kg, 8.5 mg / kg, 9.0 mg / kg, 9.5 mg / kg, 10.0 mg / kg, 10.5 mg / kg, 11.0 mg / kg, 11.5 mg / kg, or 12 mg / kg.

[0195] In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 0.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 1.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 1.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 2.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 2.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 3.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 3.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 4.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 4.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 5.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 5.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 6.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 6.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 7.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 7.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 8.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 8.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 9.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 9.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 10.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 10.5 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 11.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 11.5 mg / kg.In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 12.0 mg / kg. In some embodiments, the dose of an anti-IL-2 antibody disclosed herein is 12.5 mg / kg.

[0196] In some embodiments, the dose of IL-2 comprises a low dose. Those skilled in the art will appreciate that a low dose of IL-2 may encompass a dose level lower than that provided in tests currently known in the art. In certain embodiments, a dose of about 10×10 3 IU / kg ~ 300 x 10 3 The IL-2 dose of IU / kg encompasses a low dose of IL-2. In certain embodiments, about 10 x 10 3 IU / kg ~ 500 x 10 3 The IL-2 dose of IU / kg encompasses low doses of IL-2.

[0197] In some embodiments, the dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 IU / kg. In some embodiments, the dose of IL-2 is about 10×10 3 IU / kg ~ 300 x 10 3 IU / kg. In some embodiments, the dose of IL-2 is about 15×10 3 IU / kg ~ 270 x 10 3 In some embodiments, the dose of IL-2 is about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, or 500×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 15×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 45×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 135×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 270×10 3IU / kg. In some embodiments, the dose of IL-2 is about 300×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 400×10 3 IU / kg. In some embodiments, the dose of IL-2 is about 500×10 3 IU / kg. In some embodiments, the dose of IL-2 is 15×10 3 IU / kg. In some embodiments, the dose of IL-2 is 45×10 3 IU / kg. In some embodiments, the dose of IL-2 is 135×10 3 IU / kg. In some embodiments, the dose of IL-2 is 270×10 3 IU / kg. In some embodiments, the dose of IL-2 is 300×10 3 IU / kg. In some embodiments, the dose of IL-2 is 400×10 3 IU / kg. In some embodiments, the dose of IL-2 is 500×10 3 IU / kg.

[0198] In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is about 0.1 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 300 x 10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is about 0.1 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 45×103 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0199] In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 270×10 3In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0200] In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 400×10 3In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0201] In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0202] In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when a single loading dose of an anti-IL-2 antibody disclosed herein and IL-2 is administered, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 250×10 3 IU / kg.

[0203] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is about 0.1 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 300 x 10 3In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is about 0.1 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is about 0.5 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 300 x 10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is about 0.5 mg / kg to 12 mg / kg anti-IL-2 antibody and the dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 IU / kg.

[0204] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 300×103 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 0.5 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0205] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 1.5 mg / kg and the dose of IL-2 is 500×10 3IU / kg.

[0206] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is about 10×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is about 10×10 3 IU / kg~500IU / kg.

[0207] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 400×10 3In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 4.5 mg / kg and the dose of IL-2 is 500×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is about 10×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is about 10×10 3 IU / kg~500IU / kg.

[0208] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 400×103 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 9.0 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0209] In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 15×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 45×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 135×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 270×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 300×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 400×10 3 In some embodiments, when administering a combination therapy comprising an anti-IL-2 antibody disclosed herein and IL-2, the dose of the anti-IL-2 antibody is 12 mg / kg and the dose of IL-2 is 500×10 3 IU / kg.

[0210] In some embodiments, the dose of IL-2 is considered low when compared to other therapies. In some embodiments, the low dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 IU / kg. In some embodiments, the low dose of IL-2 is about 10×10 3 IU / kg ~ 500 x 10 3 IU / kg. In some embodiments, the low dose of IL-2 is about 15×10 3 IU / kg ~ 500 x 10 3 IU / kg. In some embodiments, the low dose of IL-2 is about 45×10 3 IU / kg ~ 500 x 10 3 In some embodiments, the low dose of IL-2 is about 500×10 3 IU / kg or less. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.014. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.023. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.038. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.043. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.053. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.054. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.066. In some embodiments, the composition comprises an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.067. In some embodiments, the composition comprises an anti-IL-2 antibody comprising the anti-IL-2 clone BDG 17.069.

[0211] In some embodiments, the composition comprises an anti-IL2 antibody and a pharmaceutically acceptable carrier. In some embodiments, the composition comprises an anti-IL2 antibody and IL-2 and a pharmaceutically acceptable carrier. In some embodiments, the composition comprises an anti-IL2 antibody conjugated to IL-2 and a pharmaceutically acceptable carrier.

[0212] In some embodiments, the anti-IL-2 antibody and IL-2 are contained in the same composition. In some embodiments, the anti-IL-2 antibody and IL-2 are contained in different compositions. In some embodiments, the administration of a combination of an anti-IL-2 antibody and IL-2, or a composition thereof, is simultaneous. In some embodiments, the administration of a combination of an anti-IL-2 antibody and IL-2, or a composition thereof, comprises administration of the anti-IL-2 antibody or a composition thereof before IL-2 or a composition thereof. In some embodiments, the administration of a combination of an anti-IL-2 antibody and IL-2, or a composition thereof, comprises administration of the anti-IL-2 antibody or a composition thereof after administration of IL-2 or a composition thereof.

[0213] Those skilled in the art will understand that a "pharmaceutical composition" can include a preparation of one or more of the active ingredients described herein with other chemical components, such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of an active agent, such as, but not limited to, an antibody or compound, to an organism.

[0214] In some embodiments, disclosed herein are pharmaceutical compositions for therapeutic use in treating a subject with a weakened immune system. In some embodiments, disclosed herein are pharmaceutical compositions for therapeutic use in treating a subject suffering from a viral infection, a bacterial infection, or cancer. In some embodiments, disclosed herein are pharmaceutical compositions for use as part of a combination therapy for treating a subject with a weakened immune system. In some embodiments, disclosed herein are pharmaceutical compositions for use as part of a combination therapy for use in treating a subject suffering from a viral infection, a bacterial infection, or cancer.

[0215] Those skilled in the art will understand that the phrases "physiologically acceptable carrier," "pharmaceutically acceptable carrier," "physiologically acceptable excipient," and "pharmaceutically acceptable excipient" can be used interchangeably and can include a carrier, excipient, or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered active ingredient.

[0216] Those skilled in the art will understand that "excipients" can include inert substances added to a pharmaceutical composition to further facilitate administration of an active ingredient. In some embodiments, excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0217] Techniques for drug formulation and administration are found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, latest edition, which is incorporated herein by reference.

[0218] In some embodiments, the compositions disclosed herein comprise therapeutic compositions. In some embodiments, the compositions disclosed herein comprise therapeutic benefits.

[0219] Combination therapy In some embodiments, the anti-IL-2 antibodies or compositions thereof disclosed herein are used as part of a combination therapy. In some embodiments, the anti-IL-2 antibodies or compositions thereof disclosed herein are used in combination with IL-2. In some embodiments, the anti-IL-2 antibodies or compositions thereof disclosed herein are used in combination with IL-2 and an immune checkpoint inhibitor. In some embodiments, the anti-IL-2 antibodies or compositions thereof disclosed herein are used in combination with an immune checkpoint inhibitor.

[0220] In some embodiments, an anti-IL-2 antibody or composition thereof is used in combination with an immune checkpoint inhibitor. In some embodiments, the term "immune checkpoint inhibitor" may encompass any compound or molecule capable of inhibiting the function of a checkpoint protein. In some embodiments, the term "immune checkpoint inhibitor" may encompass any compound or molecule that targets an immune checkpoint protein. Those skilled in the art will appreciate that "immune checkpoints" are important regulators of the immune system that, when stimulated, can attenuate an immune response to an immunological stimulus. Checkpoint inhibitors can block inhibitory checkpoints, thereby restoring immune system function. In some embodiments, the one or more checkpoint inhibitors comprise an immune checkpoint inhibitor.

[0221] Those skilled in the art will understand that terms such as "immune checkpoint inhibitor" (ICI), "checkpoint inhibitor," and the like, may be used interchangeably herein, all with the same quality and meaning, and that immune checkpoint inhibitors encompass compounds that inhibit the activity or regulatory mechanisms of the immune system. Immune system checkpoints or immune checkpoints generally are inhibitory pathways of the immune system that act to maintain self-tolerance or to regulate the duration and amplitude of physiological immune responses to minimize collateral tissue damage. Checkpoint inhibitors can inhibit immune system checkpoints by inhibiting the activity of proteins in the pathway.

[0222] Targets of immune checkpoint inhibitors include, but are not limited to, PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, and VTCN-1. In some embodiments, anti-IL-2 antibody therapy is used in combination with an immune checkpoint inhibitor, and the targets of the immune checkpoint inhibitor include PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, or any combination thereof.

[0223] Checkpoint inhibitors may include antibodies or antigen-binding fragments thereof, other binding proteins, biotherapeutics, or small molecules that bind to and block or inhibit the activity of one or more of PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1. Exemplary checkpoint inhibitors include, but are not limited to, those listed in Table 1 below.

[0224] [Table 1-1] [Table 1-2] [Table 1-3]

[0225] In some embodiments, the checkpoint inhibitor comprises a PD-1 inhibitor. In some embodiments, the checkpoint inhibitor comprises a PD-L1 inhibitor. In some embodiments, the checkpoint inhibitor comprises a CTLA-4 inhibitor. In some embodiments, the checkpoint inhibitor comprises a TIGIT inhibitor. In some embodiments, the checkpoint inhibitor comprises a TIM-3 inhibitor. In some embodiments, the checkpoint inhibitor comprises a B7-H3 inhibitor. In some embodiments, the checkpoint inhibitor comprises a CD73 inhibitor. In some embodiments, the checkpoint inhibitor comprises a LAG3 inhibitor. In some embodiments, the checkpoint inhibitor comprises a CD27 inhibitor. In some embodiments, the checkpoint inhibitor comprises a CD70 inhibitor. In some embodiments, the checkpoint inhibitor comprises a 4-1BB agonist binding agent. In some embodiments, the checkpoint inhibitor comprises a GITR agonist binding agent. In some embodiments, the checkpoint inhibitor comprises an OX40 agonist binding agent. In some embodiments, the checkpoint inhibitor comprises a SIRP-alpha (CD47) inhibitor. In some embodiments, the checkpoint inhibitor comprises a CD39 inhibitor. In some embodiments, the checkpoint inhibitor comprises an ILDR2 inhibitor. In some embodiments, the checkpoint inhibitor comprises a VISTA inhibitor. In some embodiments, the checkpoint inhibitor comprises a BTLA inhibitor. In some embodiments, the checkpoint inhibitor comprises a VTCN-1 inhibitor.

[0226] In some embodiments, the checkpoint inhibitors comprise a combination of a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, and a VTCN-1 inhibitor. In some embodiments, the checkpoint inhibitors comprise at least two checkpoint inhibitors selected from a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, and a VTCN-1 inhibitor.

[0227] In some embodiments, pharmaceutical compositions for use in the combination therapies described herein comprise an effective amount of a checkpoint inhibitor described herein and a pharmaceutically acceptable carrier.

[0228] In some embodiments, the compositions disclosed herein comprise a checkpoint inhibitor and a pharmaceutically acceptable carrier. In some embodiments, the compositions disclosed herein comprise a combination of checkpoint inhibitors and a pharmaceutically acceptable carrier. In some embodiments, the compositions comprise a checkpoint inhibitor, including a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, or a VTCN-1 inhibitor. In some embodiments, the checkpoint inhibitors comprise at least two checkpoint inhibitors chosen from a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, or a VTCN-1 inhibitor; and a pharmaceutically acceptable carrier. In some embodiments, the composition comprises at least two checkpoint inhibitors selected from a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, and a VTCN-1 inhibitor.In some embodiments, the checkpoint inhibitors comprise at least two checkpoint inhibitors selected from a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, and a VTCN-1 inhibitor; and a pharmaceutically acceptable carrier.

[0229] In certain embodiments, when two or more checkpoint inhibitors are used in the therapeutic methods described herein, each checkpoint inhibitor is included in a separate composition. In certain embodiments, when two or more checkpoint inhibitors are used in the therapeutic methods described herein, the checkpoint inhibitors can be included in the same composition.

[0230] In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof described herein and a checkpoint inhibitor or composition thereof. In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof described herein and IL-2, and a checkpoint inhibitor or composition thereof. In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof conjugated to IL-2 described herein and a checkpoint inhibitor or composition thereof.

[0231] In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof described herein and at least two checkpoint inhibitors or compositions thereof. In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof described herein and IL-2, and at least two checkpoint inhibitors or compositions thereof. In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody or composition thereof conjugated to IL-2 described herein and at least two checkpoint inhibitors or compositions thereof.

[0232] In some embodiments, the combination therapy includes a second composition comprising one or more checkpoint inhibitors described herein.

[0233] In some embodiments, the combination therapy comprises the use of an anti-IL-2 antibody BDG17.069 or a composition thereof described herein; and IL-2; and a checkpoint inhibitor or a composition thereof described herein. In some embodiments, the combination therapy comprises the use of BDG17.069 or a composition thereof described herein; and low-dose IL-2; and a checkpoint inhibitor or a composition thereof described herein. In some embodiments, the combination therapy comprises the use of BDG17.069 or a composition thereof described herein and low-dose IL-2; and a checkpoint inhibitor or a composition thereof described herein, wherein the checkpoint inhibitor is selected from a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, a TIM-3 inhibitor, a B7-H3 inhibitor, a CD73 inhibitor, a LAG3 inhibitor, a CD27 inhibitor, a CD70 inhibitor, a 4-1BB inhibitor, a GITR inhibitor, an OX40 inhibitor, a SIRP-alpha (CD47) inhibitor, a CD39 inhibitor, an ILDR2 inhibitor, a VISTA inhibitor, a BTLA inhibitor, and a VTCN-1 inhibitor.

[0234] In some embodiments, the combination therapy comprises the use of BDG17.069 or a composition thereof and low-dose IL-2 as described herein; and a checkpoint inhibitor or composition thereof, wherein the checkpoint inhibitor comprises PD-L1. In some embodiments, the combination therapy comprises the use of BDG17.069 or a composition thereof; and low-dose IL-2 (aldesleukin); and avelumab or a composition thereof. In certain embodiments of the combination therapy comprising BDG17.069 or a composition thereof and low-dose IL-2 (aldesleukin); and avelumab or a composition thereof, the IL-2 is administered by subcutaneous injection.

[0235] In some embodiments of the combination therapy, the administered IL-2 comprises a low dose of IL-2. In some embodiments of the combination therapy, the administered IL-2 is administered subcutaneously. In some embodiments of the combination therapy, the administered IL-2 comprises a low dose of IL-2 administered subcutaneously.

[0236] In some embodiments of the combination therapy, the anti-IL-2 antibody and IL-2 are in the same composition as the checkpoint inhibitor. In some embodiments, the anti-IL-2 antibody and IL-2 are in different compositions from each other and from the checkpoint inhibitor. In some embodiments, the anti-IL-2 antibody, IL-2, and checkpoint inhibitor are in the same composition. In some embodiments, the anti-IL-2 antibody and IL-2 are in one composition and the checkpoint inhibitor is in a different composition. In some embodiments, the anti-IL-2 antibody and the checkpoint inhibitor are in one composition and the IL-2 is in a different composition.

[0237] In some combination therapy embodiments, BDG17.069 and aldesleukin are included in the same composition as the PD-L1 checkpoint inhibitor. In some combination therapy embodiments, BDG17.069 and aldesleukin are included in the same composition as avelumab. In some embodiments, BDG17.069 and aldesleukin are included in different compositions from each other and avelumab. In some embodiments, BDG17.069, aldesleukin, and avelumab are included in the same composition. In some embodiments, BDG17.069 and aldesleukin are included in one composition and avelumab is included in a different composition. In some embodiments, BDG17.069 and avelumab are included in one composition and aldesleukin is included in a different composition. In some combination therapy embodiments, the anti-IL-2 antibody or composition thereof and the checkpoint inhibitor or composition thereof can be administered in any order. In some combination therapy embodiments, the BDG17.069 or composition thereof and avelumab or composition thereof can be administered in any order. In some embodiments of the combination therapy, the anti-IL-2 antibody or composition thereof, IL-2 or composition thereof, and checkpoint inhibitor or composition thereof may be administered in any order. In some embodiments of the combination therapy, the BDG17.069 or composition thereof, aldesleukin or composition thereof, and avelumab or composition thereof may be administered in any order. For example, but not limited to, the anti-IL-2 antibody may be administered before, simultaneously with, or after the administration of the checkpoint inhibitor. Similarly, the combination of the anti-IL-2 antibody and IL-2 may be administered before, simultaneously with, or after the administration of the checkpoint inhibitor. For example, but not limited to, BDG17.069 may be administered before, simultaneously with, or after the administration of avelumab. Similarly, the combination of BDG17.069 and aldesleukin may be administered before, simultaneously, or after the administration of avelumab. In some embodiments, the anti-IL-2 antibody may be administered before, simultaneously, or after the administration of at least two checkpoint inhibitors. Similarly, the combination of anti-IL-2 antibody and IL-2 can be administered before, simultaneously with, or after the administration of at least two checkpoint inhibitors.

[0238] In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises co-administration of an anti-IL-2 antibody or composition thereof and a checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises co-administration of BDG17.069 or a composition thereof and avelumab. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises co-administration of an anti-IL-2 antibody and IL-2, or a composition thereof, and a checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises co-administration of BDG17.069 and aldesleukin, or a composition thereof, and avelumab. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises pre-administration of an anti-IL-2 antibody or composition thereof before the checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises pre-administration of BDG17.069 or a composition thereof before avelumab. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises pre-administration of an anti-IL-2 antibody and IL-2, or a composition thereof, before the checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises prior administration of BDG17.069 and aldesleukin, or a composition thereof, before avelumab. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises subsequent administration of an anti-IL-2 antibody, or a composition thereof, after administration of the checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises subsequent administration of BDG17.069, or a composition thereof, after administration of avelumab. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises subsequent administration of an anti-IL-2 antibody and IL-2, or a composition thereof, after administration of the checkpoint inhibitor. In some embodiments, administering the combination therapy with a checkpoint inhibitor comprises subsequent administration of BDG17.069 and aldesleukin, or a composition thereof, after administration of avelumab.

[0239] In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising heavy and light chain variable regions having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27, or SEQ ID NOs: 36 and 37. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising a heavy chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 38-40, respectively; 44-46, respectively; 50-52, respectively; 56-58, respectively; or 62-64, respectively. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising a light chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 41-43, respectively; 47-49, respectively; 53-55, respectively; 59-61, respectively; or 65-67, respectively. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising a heavy chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 38-40, respectively; SEQ ID NOs: 44-46, respectively; SEQ ID NOs: 50-52, respectively; SEQ ID NOs: 56-58, respectively; or SEQ ID NOs: 62-64, respectively, and a light chain variable domain comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences of SEQ ID NOs: 41-43, respectively; SEQ ID NOs: 47-49, respectively; SEQ ID NOs: 53-55, respectively; SEQ ID NOs: 59-61, respectively; or SEQ ID NOs: 65-67, respectively.

[0240] In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising any of clones BDG 17.014, BDG 17.023, BDG 17.038, BDG 17.043, BDG 17.053, BDG 17.054, BDG 17.066, BDG 17.067, and BDG 17.069. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.014. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.023. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody comprising anti-IL-2 clone BDG 17.038. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.043. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.053. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.054. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.066. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.067. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.069.

[0241] In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including any of clones BDG 17.014, BDG 17.023, BDG 17.038, BDG 17.043, BDG 17.053, BDG 17.054, BDG 17.066, BDG 17.067, and BDG 17.069, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.014, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.023, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor and an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.038, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.043, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.053, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.054, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.066, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.067, and IL-2. In some embodiments, the combination therapy comprises the use of a checkpoint inhibitor, an anti-IL-2 antibody, including anti-IL-2 clone BDG 17.069, and IL-2.

[0242] In certain embodiments, the use of the combination therapy is to treat a cancer or tumor, hi some embodiments, the use of the combination therapy is to treat a solid cancer or solid tumor described herein.

[0243] In some embodiments of the combination therapies disclosed herein, treating a solid tumor includes treating the primary tumor and secondary metastases of the tumor. In some embodiments of the combination therapies disclosed herein, treating a solid tumor includes treating secondary metastases of the tumor. In some embodiments of the combination therapies disclosed herein, treating a solid tumor includes second-line treatment of the tumor. In some embodiments of the combination therapies disclosed herein, treating a solid tumor includes third-line treatment of the tumor. In some embodiments of the combination therapies disclosed herein, treating a solid tumor includes second-line and third-line treatment of the tumor.

[0244] In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises a metastatic cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises an unresectable locally advanced cancer or tumor. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises a metastasis. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises an unresectable locally advanced cancer or tumor.

[0245] In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises head and neck cancer, head and neck squamous cell carcinoma (HNSCC), pancreatic cancer, lung cancer, thyroid cancer, non-small cell lung cancer (NSCLC), nasopharyngeal carcinoma, melanoma, acral melanoma, uveal malignant melanoma, colorectal cancer (CRC), bladder cancer, bile duct cancer (cholangiocarcinoma), uterine cancer, cervical cancer, gallbladder cancer, or renal cell carcinoma (RCC). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises urothelial carcinoma, adrenocortical carcinoma, clear cell renal cell carcinoma (ccRCC), melanoma, triple-negative breast cancer, head and neck squamous cell carcinoma (HNSCC), gastric or gastroesophageal cancer, esophageal squamous cell carcinoma, cutaneous squamous cell carcinoma (cSCC), pancreatic adenocarcinoma, cholangiocarcinoma, hepatocellular carcinoma (HCC), colorectal cancer (CRC), epithelial ovarian cancer, cervical cancer, endometrial cancer, thyroid cancer (follicular or papillary histology), non-small cell lung cancer (NSCLC), or Merkel cell carcinoma. In some embodiments, the urothelial carcinoma arises in the bladder, renal pelvis, ureter, or urethra, or any combination thereof. In certain embodiments, the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the urothelial cancer occurs in the bladder, renal pelvis, ureter, or urethra, or any combination thereof, and the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the adrenocortical carcinoma includes unresectable, locally advanced, or metastatic cancer. In some embodiments, the clear cell renal cell carcinoma (ccRCC) includes cancer that has progressed during or after at least two approved treatment regimens (e.g., small molecule inhibitors, anti-PDx therapy). In some embodiments, the melanoma includes cancer that is either locally unresectable or metastatic, and the locally unresectable or metastatic cancer may include: (a) BRAF wt: the cancer progressed after receiving anti-PD-1-containing therapy with or without anti-CTLA-4; or (b) BRAF mut: the cancer progressed after a BRAF+MEK inhibitor.In some embodiments, triple-negative breast cancer includes cancers that are unresectable, locally advanced, or metastatic and refractory to standard first-line therapy (which may include, for example, but is not limited to, cytotoxic chemotherapy alone and / or poly ADP-ribose polymerase (PARP) inhibitors for breast cancer gene (BRCA) 1 or 2 mutations, and / or anti-PDx therapy in MSI-H / dMMR-positive tumors). In some embodiments, head and neck squamous cell carcinoma (HNSCC) includes, for example, but is not limited to, cancers that progress during or after treatment with anti-PDx for metastatic or recurrent disease (unless ineligible, e.g., patients who have failed chemotherapy and have a PD-L1 combined positive score (CPS)<1) and platinum-based chemotherapy (unless ineligible for platinum chemotherapy). In some embodiments, gastric or gastroesophageal cancer includes cancers that progress during or after cytotoxic chemotherapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents) with or without trastuzumab (for HER2-overexpressing adenocarcinoma) and with or without anti-PD-1 inhibitor therapy. Patients with a CPS of ≥ 1 may have received an anti-PD-1 containing regimen (unless intolerant or treatment is unavailable). In some embodiments, esophageal squamous cell carcinoma includes cancers that progress during or after cytotoxic chemotherapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents) with anti-PD-1 therapy. Patients with a CPS of ≥ 10 may have received an anti-PD-1 containing regimen (unless intolerant or treatment is unavailable). In some embodiments, cutaneous squamous cell carcinoma (cSCC) includes recurrent or metastatic cSCC that is incurable by surgery or radiation. In some embodiments, pancreatic adenocarcinoma includes cancers that are unresectable, locally advanced, or metastatic and have received at least first-line chemotherapy (e.g., but not limited to, FOLFIRINOX; unless ineligible or infeasible). In some embodiments, cholangiocarcinoma includes unresectable, locally advanced, or metastatic cancers in patients who may have received first-line or more systemic chemotherapy, unless the patient is ineligible for chemotherapy. In some embodiments, hepatocellular carcinoma (HCC) includes cancers that progress during or after an approved treatment regimen (unless ineligible).In some embodiments, colorectal cancer (CRC) includes: (a) K-Ras wild-type: patients who have progressed during or after, or are ineligible for, both irinotecan-based chemotherapy and oxaliplatin-based chemotherapy, and who have relapsed or are refractory to at least one prior systemic therapy, including an anti-epidermal growth factor receptor (EGFR) antibody, such as cetuximab or panitumumab; or (b) K-Ras mutant: patients who have progressed during or after, or are ineligible for, irinotecan-based chemotherapy and oxaliplatin-based chemotherapy (± bevacizumab). In some embodiments, epithelial ovarian cancer includes cancers that progress during or after at least one prior cytotoxic chemotherapy regimen (unless ineligible) followed by poly ADP-ribose polymerase (PARP) inhibitor therapy (unless ineligible) in BRCA mutation-positive patients. In some embodiments, cervical cancer includes cancers that progress during or after first-line cytotoxic chemotherapy and second-line cytotoxic chemotherapy or anti-PDx therapy (unless ineligible) in PD-L1-positive (CPS≧1) or MSI-H / dMMR-positive tumors. In some embodiments, the patient's endometrial cancer includes cancers that progress during or after cytotoxic chemotherapy (e.g., without limitation, ±trastuzumab) or hormonal therapy, and anti-PDx therapy in MSI-H / dMMR-positive tumors. In some embodiments, thyroid cancer (follicular or papillary histology) includes cancers that are iodine-refractory. In some embodiments, non-small cell lung cancer (NSCLC) includes cancers that progress during or after treatment with platinum-based chemotherapy and anti-PDx therapy for unresectable, locally advanced, or metastatic disease. In some embodiments, NSCLC with activating EGFR mutations (excluding exon 20 insertion mutations) or anaplastic lymphoma kinase (ALK) rearrangements must have progressed following treatment with platinum-based chemotherapy (unless ineligible for platinum therapy) plus available EGFR- or ALK-targeted therapy. In some embodiments, Merkel cell carcinoma includes metastatic Merkel cell carcinoma that is incurable by surgery or radiation.

[0246] In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises head and neck cancer, pancreatic cancer, or non-small cell lung cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises non-small cell lung cancer (NSCLC), melanoma, metastatic melanoma, primary and metastatic melanoma, cutaneous squamous cell carcinoma, or renal cell carcinoma (RCC). In some embodiments of the combination therapies disclosed herein, the melanoma comprises acral melanoma or uveal malignant melanoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises head and neck cancer. In some embodiments, the solid tumor comprises pancreatic cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises lung cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises thyroid cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises non-small cell lung cancer (NSCLC). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises nasopharyngeal carcinoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises melanoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises acral melanoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises uveal malignant melanoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises colorectal cancer (CRC). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises bladder cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises bile duct carcinoma (cholangiocarcinoma). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises uterine cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises cervical cancer. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises gallbladder cancer. In some embodiments of the combination therapy disclosed herein, the solid tumor being treated comprises renal cell carcinoma (RCC).In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises head and neck cancer, wherein the head and neck cancer is head and neck squamous cell carcinoma (HNSCC). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises CRC, wherein the CRC has high MSI. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises melanoma, wherein the melanoma has a wild-type BRAF gene or a mutated BRAF gene. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises pancreatic cancer, wherein the pancreatic cancer is adenocarcinoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises non-small cell lung cancer (NSCLC), wherein the NSCLC is squamous cell carcinoma. In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises NSCLC, wherein the NSCLC has a mutant epidermal growth factor receptor (EGFRm). In some embodiments of the combination therapies disclosed herein, the solid tumor being treated comprises cutaneous squamous cell carcinoma.

[0247] formulation Pharmaceutical compositions disclosed herein containing anti-IL-2 antibodies, or combinations of anti-IL-2 antibodies and IL-2, or checkpoint inhibitors can be conveniently provided as sterile liquid preparations, such as isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions that can be buffered to a selected pH. Liquid preparations are typically easier to prepare than gels, other viscous compositions, and solid compositions. Furthermore, liquid compositions are somewhat more convenient to administer, particularly by injection. Viscous compositions, on the other hand, can be formulated within an appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can contain a carrier, which can be a solvent or dispersion medium containing, for example, water, saline, phosphate-buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof.

[0248] Sterile injectable solutions can be prepared by incorporating the anti-IL-2 antibody, or anti-IL-2 antibody and IL-2 combination, or checkpoint inhibitor described herein and utilized in practicing the methods disclosed herein, in the required amount of an appropriate solvent, containing various amounts of other ingredients, as needed. Such formulations may be mixed with a suitable carrier, diluent, or excipient, such as sterile water, saline, glucose, dextrose, or the like. The formulations can also be lyophilized. Depending on the route of administration and the desired preparation, the formulations may contain auxiliary substances, such as wetting agents, dispersing agents, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or thickening additives, preservatives, flavoring agents, coloring agents, and the like. Suitable formulations can be prepared without undue experimentation by reference to standard texts, such as "REMINGTON'S PHARMACEUTICAL SCIENCE," 17th Edition, 1985, incorporated herein by reference.

[0249] Various additives can be added to enhance the stability and sterility of the formulation, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0250] In certain embodiments, the terms "pharmaceutical composition," "composition," and "formulation" may be used interchangeably with the same meaning and quality.

[0251] The compositions or formulations described herein can be isotonic, i.e., they can have the same osmotic pressure as blood and tears.The desired isotonicity of the compositions disclosed herein can be achieved using sodium chloride or other pharmaceutically acceptable agents such as dextrose, boric acid, sodium tartrate, propylene glycol, or other inorganic or organic solutes.Sodium chloride may be preferred, especially for buffers containing sodium ions.

[0252] The viscosity of the composition can be maintained at a selected level, if desired, using a pharmaceutically acceptable thickening agent. Methylcellulose may be preferred because it is readily and economically available and easy to handle.

[0253] Other suitable thickening agents include, for example, xanthan gum, carboxymethylcellulose, hydroxypropylcellulose, carbomer, etc. The preferred concentration of the thickening agent depends on the agent selected. The key is to use an amount that achieves the selected viscosity. Obviously, the selection of suitable carriers and other additives will depend on the exact route of administration and the nature of the particular dosage form, e.g., liquid dosage form (e.g., whether the composition is formulated as a solution, suspension, gel, or another liquid form, e.g., time-release form or liquid-fill form).

[0254] In some embodiments, the composition is formulated to have a pH of about pH 5.0-6.0. In some embodiments, the composition is formulated to have a pH of about pH 5.0-7.0. In some embodiments, the composition is formulated to have a pH of about pH 5.0-6.5. In some embodiments, the composition is formulated to have a pH of about pH 5.0-5.5. In some embodiments, the composition is formulated to have a pH of about pH 5.5-6.0. In some embodiments, the composition is formulated to have a pH of about pH 5.5-6.5. In some embodiments, the composition is formulated to have a pH of about pH 5.0. In some embodiments, the composition is formulated to have a pH of about pH 5.5. In some embodiments, the composition is formulated to have a pH of about pH 6.0. In some embodiments, the composition is formulated to have a pH of about pH 6.5.

[0255] In some embodiments, the composition is formulated to have a pH of about pH 5.0-6.0 and comprises a buffer. In some embodiments, the buffer comprises a pharmaceutically acceptable buffer. In some embodiments, the buffer comprises a histidine buffer or a citrate buffer. In some embodiments, the buffer comprises a histidine buffer. In some embodiments, the buffer comprises a citrate buffer.

[0256] In some embodiments, the composition is formulated to have a pH of about 5.0-6.0 and comprises a buffer selected from a histidine buffer and a citrate buffer. In some embodiments, the composition is formulated to have a pH of about 5.0-6.0 and comprises a histidine buffer. In some embodiments, the composition is formulated to have a pH of about 5.0-6.0 and comprises a citrate buffer.

[0257] In some embodiments, the composition further comprises at least one of sucrose, methionine, or PS80, or any combination thereof. In some embodiments, the composition further comprises sucrose. In some embodiments, the composition further comprises methionine. In some embodiments, the composition further comprises PS80.

[0258] In some embodiments, a composition comprises an anti-IL-2 antibody disclosed herein, formulated to a pH of about pH 5.0 to 6.0, and comprising a buffer selected from a histidine buffer and a citrate buffer, and further comprising IL-2.

[0259] Those skilled in the art will recognize that the components of the composition or formulation should be selected to be chemically inert and not affect the viability or efficacy of the early apoptotic cell populations described herein for use in the methods disclosed herein, which will not pose a problem to one skilled in the art of chemical and pharmaceutical principles or which can be easily circumvented by reference to standard texts or by simple experimentation (without undue experimentation) from this disclosure and the literature cited herein.

[0260] How to use In one embodiment, the present disclosure provides a method for producing a heavy chain variable region of an anti-IL-2 antibody, the method comprising culturing a host cell under conditions conducive to expression of a vector encoding the heavy chain variable region, thereby producing the heavy chain variable region of the anti-IL-2 antibody.

[0261] In one embodiment, the present disclosure provides a method for producing a light chain variable region of an anti-IL-2 antibody, the method comprising culturing a host cell under conditions conducive to expression of a vector encoding the light chain variable region, thereby producing the light chain variable region of the anti-IL-2 antibody.

[0262] The VH and / or VL polypeptides disclosed herein can be used in therapeutic methods. In one embodiment, the polypeptides of the present disclosure can be used as immunotherapeutic agents, for example, for the differential activation of immune cells as described herein. The polypeptides can be administered to a subject directly or by administering a nucleic acid sequence encoding the polypeptide to a subject, which nucleic acid sequence can be carried by a vector.

[0263] The exact amount of the polypeptide or composition thereof required to induce the desired effect will vary from subject to subject, depending on the species, age, sex, weight, and general condition of the subject, the specific polypeptide, the route of administration, and whether other drugs are included in the regimen. Thus, it is not possible to specify an exact amount for every composition. However, an appropriate amount can be determined by one of ordinary skill in the art using routine experimentation. Dosages can vary, and the polypeptide can be administered in one or multiple doses (e.g., two or more, three or more, four or more, or five or more) per day for one or more days. Guidance for selecting the appropriate dose of an antibody can be readily found in the literature.

[0264] In some embodiments of the methods of using anti-IL-2 antibodies described herein, the subject comprises a mammalian subject. In some embodiments, the subject comprises a human subject. In some embodiments, the subject suffers from an immune deficiency. Treatment of an immune-deficient subject may, in some embodiments, comprise prophylactic treatment.

[0265] In one embodiment, the present disclosure provides a method of promoting differential proliferation of immune cells in a subject, the method comprising: preparing a composition comprising an anti-IL-2 antibody disclosed herein; and administering the composition to the subject, thereby promoting differential proliferation of immune cells in the subject. In one embodiment, the present disclosure provides a method of promoting differential proliferation of immune cells in a subject, the method comprising: preparing a composition comprising IL-2 and an anti-IL-2 antibody disclosed herein; and administering the composition to the subject, thereby promoting differential proliferation of immune cells in the subject. In one embodiment, the subject can be an animal or a human. In one embodiment, the immune cells are CD8 + The cells may be NK cells or NK cells.

[0266] Disclosed herein in some embodiments is a method of treating a disease or condition in a subject, the method comprising administering to the subject a composition comprising an anti-IL-2 antibody disclosed herein, wherein the antibody promotes differential proliferation of a subset of immune cells and reduces undesirable effects caused by IL-2, thereby treating the disease or condition in the subject. In some embodiments, the method of treating a disease disclosed herein comprises the use of a composition comprising an anti-IL-2 antibody and IL-2, or an anti-IL-2 antibody conjugated to IL-2. In some embodiments, the method of treating a disease comprises treating a viral infection, a bacterial infection, or cancer. In some embodiments, the method of treating a condition comprises treating a weak immune system, wherein the treatment prophylactically boosts the immune system.

[0267] In some embodiments of the methods of treating a disease or condition, the condition comprises a genetic predisposition that increases the likelihood of cancer in the subject. In some embodiments, the genetic predisposition comprises an alteration in the expression or activity of a gene product. In some embodiments, the genetic predisposition that increases the likelihood of cancer comprises a mutation in a tumor suppressor gene or a mismatch repair (MMR) gene, or a combination thereof.

[0268] Many hereditary cancers are known in the art, non-limiting examples include, but are not limited to, hereditary breast and ovarian cancer (HBOC) syndrome, Lynch syndrome (hereditary nonpolyposis colorectal cancer), and Li-Fraumeni syndrome.

[0269] In some embodiments, a genetic predisposition increases the likelihood of HBOC. HBOC is associated with mutations in the BRAC1 and BRAC2 genes. HBOC is associated with several different cancers, including, but not limited to, breast cancer, fallopian tube cancer, primary peritoneal cancer, male breast cancer, pancreatic cancer, and prostate cancer. In some embodiments, a genetic predisposition increases the likelihood of any of breast cancer, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, male breast cancer, pancreatic cancer, or prostate cancer, or a combination thereof.

[0270] In some embodiments, a genetic predisposition increases the likelihood of hereditary nonpolyposis colorectal cancer (HNPCC). HNPCC is associated with mutations in genes including, but not limited to, MLH1, MSH2, MSH6, PMS1, and PMS2. HNPCC is associated with a higher risk of developing endometrial cancer and cancers of the ovaries, stomach, small intestine, pancreas, kidney, brain, ureter, and bile duct. In some embodiments, a genetic predisposition increases the likelihood of any of hereditary nonpolyposis colorectal cancer, ovarian cancer, stomach cancer, small intestine cancer, pancreatic cancer, kidney cancer, brain cancer, ureter cancer, and bile duct cancer.

[0271] In some embodiments, the genetic predisposition increases the likelihood of Li-Fraumeni syndrome. Li-Fraumeni syndrome is associated with cancer, including sarcoma, osteosarcoma, soft tissue sarcoma, leukemia, brain (central nervous system) cancer, adrenocortical carcinoma, and breast cancer, or a combination thereof. In some embodiments, the genetic predisposition increases the likelihood of sarcoma, osteosarcoma, soft tissue sarcoma, leukemia, brain (central nervous system) cancer, adrenocortical carcinoma, and breast cancer, or a combination thereof.

[0272] The anti-IL2 antibodies described and exemplified herein bind to the portion of IL-2 that interacts with the alpha (CD25) receptor subunit, a component of the IL-2 trimeric receptor (CD25 / CD132 / CD122, sometimes designated as α / β / γ) found on Treg cells, eosinophils, and lung and vascular endothelial cells. In some embodiments, the anti-IL2 antibodies disclosed herein prevent activation of the trimeric IL-2 receptor found on Treg cells, eosinophils, and lung and vascular endothelial cells. In some embodiments, the anti-IL2 antibodies disclosed herein bind to IL-2 and activate signaling through the IL-2 dimeric receptor (CD132 / CD122, sometimes designated as β / γ) found on naive Teff cells, NK cells, and natural killer T (NKT) cells.

[0273] In some embodiments, the condition treated in the subject comprises treating a subject with a genetic predisposition comprising an alteration in the expression or activity of a gene product, wherein the gene comprises BRCA1, BRAC2, MLH1, MSH2, MSH6, PMS1, PMS2, TP53, or CHEK2, or a combination thereof.

[0274] As described herein, conjugates of IL-2 and the disclosed anti-IL-2 antibodies induce memory phenotype effector T cells (MP) CD8 + showed a significant effect in inducing proliferation of CD4 cells and NK cells, but +The effect on Tregs was much less. Thus, the modified anti-IL-2 antibodies disclosed herein may be useful in modulating immune cell populations and inducing differential expansion of certain immune effector cells. In one embodiment, such differential expansion of immune effector cells results in strong activation of the immune system and may be useful in treating tumors.

[0275] In some embodiments, the treatment includes treating a solid tumor. In some embodiments, the treatment includes treating a non-solid tumor. In some embodiments, the treatment includes treating a solid tumor and / or a non-solid tumor, such as, but not limited to, melanoma, renal cell carcinoma, small cell lung cancer, or other cancerous conditions. In another embodiment, the methods disclosed herein may be useful for treating a viral or bacterial infection. In another embodiment, the methods disclosed herein may be useful for treating or preventing a condition caused by IL-2 binding to endothelial CD25-expressing cells, e.g., pulmonary edema or IL-2-induced vascular leakage.

[0276] In some embodiments, the described methods of use for treating a disease or condition treat a solid tumor. In some embodiments, the solid tumor comprises head and neck cancer, head and neck squamous cell carcinoma (HNSCC), pancreatic cancer, lung cancer, thyroid cancer, non-small cell lung cancer (NSCLC), nasopharyngeal carcinoma, melanoma, acral melanoma, uveal malignant melanoma, colorectal cancer (CRC), bladder cancer, bile duct cancer (cholangiocarcinoma), uterine cancer, cervical cancer, gallbladder cancer, cutaneous squamous cell carcinoma, or renal cell carcinoma (RCC). In some embodiments, the solid tumor comprises head and neck cancer, pancreatic cancer, or non-small cell lung cancer. In some embodiments, the solid tumor comprises non-small cell lung cancer (NSCLC), melanoma, metastatic melanoma, primary and metastatic melanoma, or renal cell carcinoma (RCC). In some embodiments, the solid tumor comprises head and neck cancer. In some embodiments, the solid tumor comprises pancreatic cancer. In some embodiments, the solid tumor comprises lung cancer. In some embodiments, the solid tumor comprises thyroid cancer. In some embodiments, the solid tumor comprises non-small cell lung cancer (NSCLC). In some embodiments, the solid tumor comprises nasopharyngeal carcinoma. In some embodiments, the solid tumor comprises melanoma. In some embodiments, the melanoma comprises acral melanoma or uveal malignant melanoma. In some embodiments, the solid tumor comprises colorectal cancer (CRC). In some embodiments, the solid tumor comprises bladder cancer. In some embodiments, the solid tumor comprises bile duct cancer (cholangiocarcinoma). In some embodiments, the solid tumor comprises uterine cancer. In some embodiments, the solid tumor comprises cervical cancer. In some embodiments, the solid tumor comprises gallbladder cancer. In some embodiments, the solid tumor comprises renal cell carcinoma (RCC). In some embodiments, the head and neck cancer is head and neck squamous cell carcinoma (HNSCC). In some embodiments, the CRC has high MSI. In some embodiments, the melanoma has a wild-type BRAF gene. In some embodiments, the melanoma has a mutated BRAF gene. In some embodiments, the pancreatic cancer is adenocarcinoma. In some embodiments, the non-small cell lung cancer (NSCLC) is squamous cell carcinoma. In some embodiments, the NSCLC has a mutant epidermal growth factor receptor (EGFRm). In some embodiments, the solid tumor comprises cutaneous squamous cell carcinoma.

[0277] In some embodiments, the method of use for treating cancer includes treating a solid tumor. In certain embodiments, the solid tumor treated comprises urothelial carcinoma, adrenocortical carcinoma, clear cell renal cell carcinoma (ccRCC), melanoma, triple-negative breast cancer, head and neck squamous cell carcinoma (HNSCC), gastric or gastroesophageal cancer, esophageal squamous cell carcinoma, cutaneous squamous cell carcinoma (cSCC), pancreatic adenocarcinoma, cholangiocarcinoma, hepatocellular carcinoma (HCC), colorectal cancer (CRC), epithelial ovarian cancer, cervical cancer, endometrial cancer, thyroid cancer (follicular or papillary histology), non-small cell lung cancer (NSCLC), or Merkel cell carcinoma. In some embodiments, the urothelial carcinoma arises in the bladder, renal pelvis, ureter, or urethra, or any combination thereof. In certain embodiments, the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the urothelial cancer occurs in the bladder, renal pelvis, ureter, or urethra, or any combination thereof, and the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the adrenocortical carcinoma includes unresectable, locally advanced, or metastatic cancer. In some embodiments, the clear cell renal cell carcinoma (ccRCC) includes cancer that has progressed during or after at least two approved treatment regimens (e.g., small molecule inhibitors, anti-PDx therapy). In some embodiments, the melanoma includes cancer that is either locally unresectable or metastatic, and the locally unresectable or metastatic cancer may include: (a) BRAF wt: the cancer progressed after receiving anti-PD-1-containing therapy with or without anti-CTLA-4; or (b) BRAF mut: the cancer progressed after a BRAF+MEK inhibitor. In some embodiments, triple-negative breast cancer includes cancers that are unresectable, locally advanced, or metastatic and refractory to standard first-line therapy (which may include, for example, but are not limited to, cytotoxic chemotherapy alone and / or poly ADP-ribose polymerase (PARP) inhibitors for breast cancer gene (BRCA) 1 or 2 mutations, and / or anti-PDx therapy in MSI-H / dMMR-positive tumors).In some embodiments, head and neck squamous cell carcinoma (HNSCC) includes, for example, but not limited to, cancers progressing during or after treatment with anti-PDx (unless ineligible, e.g., patients who have failed chemotherapy and have a PD-L1 combination positive score (CPS) < 1) and platinum-based chemotherapy (unless ineligible for platinum chemotherapy) for metastatic or recurrent disease. In some embodiments, gastric or gastroesophageal cancer includes cancers progressing during or after cytotoxic chemotherapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents) with or without trastuzumab (for HER2-overexpressing adenocarcinoma) and with or without anti-PD-1 inhibitor therapy. Patients with a CPS ≥ 1 may have received an anti-PD-1-containing regimen (unless intolerant or treatment is unavailable). In some embodiments, esophageal squamous cell carcinoma includes cancers progressing during or after cytotoxic chemotherapy with anti-PD-1 therapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents). Patients with a CPS≧10 may have received an anti-PD-1 containing regimen (unless intolerant or treatment is unavailable). In some embodiments, cutaneous squamous cell carcinoma (cSCC) includes recurrent or metastatic cSCC that is incurable by surgery or radiation. In some embodiments, pancreatic adenocarcinoma includes cancers that are unresectable, locally advanced, or metastatic and have received at least first-line chemotherapy (e.g., but not limited to, FOLFIRINOX; unless ineligible or infeasible). In some embodiments, cholangiocarcinoma includes unresectable, locally advanced, or metastatic cancers in patients who may have received first-line or more systemic chemotherapy, unless the patient is ineligible for chemotherapy. In some embodiments, hepatocellular carcinoma (HCC) includes cancers that progress during or after an approved treatment regimen (unless ineligible).In some embodiments, colorectal cancer (CRC) includes: (a) K-Ras wild-type: patients who have progressed during or after, or are ineligible for, both irinotecan-based chemotherapy and oxaliplatin-based chemotherapy, and who have relapsed or are refractory to at least one prior systemic therapy, including an anti-epidermal growth factor receptor (EGFR) antibody, such as cetuximab or panitumumab; or (b) K-Ras mutant: patients who have progressed during or after, or are ineligible for, irinotecan-based chemotherapy and oxaliplatin-based chemotherapy (± bevacizumab). In some embodiments, epithelial ovarian cancer includes cancers that progress during or after at least one prior cytotoxic chemotherapy regimen (unless ineligible) followed by poly ADP-ribose polymerase (PARP) inhibitor therapy (unless ineligible) in BRCA mutation-positive patients. In some embodiments, cervical cancer includes cancers that progress during or after first-line cytotoxic chemotherapy and second-line cytotoxic chemotherapy or anti-PDx therapy (unless ineligible) in PD-L1-positive (CPS≧1) or MSI-H / dMMR-positive tumors. In some embodiments, the patient's endometrial cancer includes cancers that progress during or after cytotoxic chemotherapy (e.g., without limitation, ±trastuzumab) or hormonal therapy, and anti-PDx therapy in MSI-H / dMMR-positive tumors. In some embodiments, thyroid cancer (follicular or papillary histology) includes cancers that are iodine-refractory. In some embodiments, non-small cell lung cancer (NSCLC) includes cancers that progress during or after treatment with platinum-based chemotherapy and anti-PDx therapy for unresectable, locally advanced, or metastatic disease. In some embodiments, NSCLC with activating EGFR mutations (excluding exon 20 insertion mutations) or anaplastic lymphoma kinase (ALK) rearrangements must have progressed following treatment with platinum-based chemotherapy (unless ineligible for platinum therapy) plus available EGFR- or ALK-targeted therapy. In some embodiments, Merkel cell carcinoma includes metastatic Merkel cell carcinoma that is incurable by surgery or radiation.

[0278] In some embodiments, treating a solid tumor includes treating the primary tumor and secondary metastases of the tumor. In some embodiments, treating a solid tumor includes treating secondary metastases of the tumor. In some embodiments, treating a solid tumor includes second line treatment of the tumor. In some embodiments, treating a solid tumor includes third line treatment of the tumor. In some embodiments, treating a solid tumor includes second and third line treatment of the tumor.

[0279] As used throughout, the terms "cancer" and "tumor" may, in some embodiments, be used interchangeably, with the same meaning and qualities.

[0280] In some embodiments, the solid tumor being treated comprises a metastatic cancer. In some embodiments, the solid tumor being treated comprises an unresectable locally advanced cancer or tumor. In some embodiments, the solid tumor being treated comprises a metastasis. In some embodiments, the solid tumor being treated comprises an unresectable locally advanced cancer or tumor.

[0281] In some embodiments, the subject treated by the methods disclosed herein has any of 19 types of solid tumors. In some embodiments, the solid tumor comprises unresectable locally advanced cancer or metastatic cancer. In some embodiments, the solid tumor comprises unresectable locally advanced cancer. In some embodiments, the solid tumor comprises metastatic cancer. In some embodiments, the subject treated by the methods disclosed herein is ineligible for treatment with standard and / or approved therapies. In some embodiments, the subject treated is human.

[0282] In some embodiments, the subject treated by the methods disclosed herein has or is afflicted with urothelial carcinoma, adrenocortical carcinoma, clear cell renal cell carcinoma (ccRCC), melanoma, triple-negative breast cancer, head and neck squamous cell carcinoma (HNSCC), gastric or gastroesophageal cancer, esophageal squamous cell carcinoma, cutaneous squamous cell carcinoma (cSCC), pancreatic adenocarcinoma, cholangiocarcinoma, hepatocellular carcinoma (HCC), colorectal cancer (CRC), epithelial ovarian cancer, cervical cancer, endometrial cancer, thyroid cancer (follicular or papillary histology), non-small cell lung cancer (NSCLC), or Merkel cell carcinoma. In some embodiments, the urothelial carcinoma arises in the bladder, renal pelvis, ureter, or urethra, or any combination thereof. In certain embodiments, the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the urothelial cancer occurs in the bladder, renal pelvis, ureter, or urethra, or any combination thereof, and the cancer has progressed during or after anti-PDx therapy and, if eligible, a platinum-containing regimen. In some embodiments, the adrenocortical carcinoma includes unresectable, locally advanced, or metastatic cancer. In some embodiments, the clear cell renal cell carcinoma (ccRCC) includes cancer that has progressed during or after at least two approved treatment regimens (e.g., small molecule inhibitors, anti-PDx therapy). In some embodiments, the melanoma includes cancer that is either locally unresectable or metastatic, and the locally unresectable or metastatic cancer may include: (a) BRAF wt: the cancer progressed after receiving anti-PD-1-containing therapy with or without anti-CTLA-4; or (b) BRAF mut: the cancer progressed after a BRAF+MEK inhibitor. In some embodiments, triple-negative breast cancer includes cancers that are unresectable, locally advanced, or metastatic and refractory to standard first-line therapy (which may include, for example, but are not limited to, cytotoxic chemotherapy alone and / or poly ADP-ribose polymerase (PARP) inhibitors for breast cancer gene (BRCA) 1 or 2 mutations, and / or anti-PDx therapy in MSI-H / dMMR-positive tumors).In some embodiments, head and neck squamous cell carcinoma (HNSCC) includes, for example, but not limited to, cancers progressing during or after treatment with anti-PDx (unless ineligible, e.g., patients who have failed chemotherapy and have a PD-L1 combination positive score (CPS) < 1) and platinum-based chemotherapy (unless ineligible for platinum chemotherapy) for metastatic or recurrent disease. In some embodiments, gastric or gastroesophageal cancer includes cancers progressing during or after cytotoxic chemotherapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents) with or without trastuzumab (for HER2-overexpressing adenocarcinoma) and with or without anti-PD-1 inhibitor therapy. Patients with a CPS ≥ 1 may have received an anti-PD-1-containing regimen (unless intolerant or treatment is unavailable). In some embodiments, esophageal squamous cell carcinoma includes cancers progressing during or after cytotoxic chemotherapy with anti-PD-1 therapy (e.g., but not limited to, paclitaxel, fluoropyrimidines, platinum agents). Patients with a CPS≧10 may have received an anti-PD-1 containing regimen (unless intolerant or treatment is unavailable). In some embodiments, cutaneous squamous cell carcinoma (cSCC) includes recurrent or metastatic cSCC that is incurable by surgery or radiation. In some embodiments, pancreatic adenocarcinoma includes cancers that are unresectable, locally advanced, or metastatic and have received at least first-line chemotherapy (e.g., but not limited to, FOLFIRINOX; unless ineligible or infeasible). In some embodiments, cholangiocarcinoma includes unresectable, locally advanced, or metastatic cancers in patients who may have received first-line or more systemic chemotherapy, unless the patient is ineligible for chemotherapy. In some embodiments, hepatocellular carcinoma (HCC) includes cancers that progress during or after an approved treatment regimen (unless ineligible).In some embodiments, colorectal cancer (CRC) includes: (a) K-Ras wild-type: patients who have progressed during or after, or are ineligible for, both irinotecan-based chemotherapy and oxaliplatin-based chemotherapy, and who have relapsed or are refractory to at least one prior systemic therapy, including an anti-epidermal growth factor receptor (EGFR) antibody, such as cetuximab or panitumumab; or (b) K-Ras mutant: patients who have progressed during or after, or are ineligible for, irinotecan-based chemotherapy and oxaliplatin-based chemotherapy (± bevacizumab). In some embodiments, epithelial ovarian cancer includes cancers that progress during or after at least one prior cytotoxic chemotherapy regimen (unless ineligible) followed by poly ADP-ribose polymerase (PARP) inhibitor therapy (unless ineligible) in BRCA mutation-positive patients. In some embodiments, cervical cancer includes cancers that progress during or after first-line cytotoxic chemotherapy and second-line cytotoxic chemotherapy or anti-PDx therapy (unless ineligible) in PD-L1-positive (CPS≧1) or MSI-H / dMMR-positive tumors. In some embodiments, the patient's endometrial cancer includes cancers that progress during or after cytotoxic chemotherapy (e.g., without limitation, ±trastuzumab) or hormonal therapy, and anti-PDx therapy in MSI-H / dMMR-positive tumors. In some embodiments, thyroid cancer (follicular or papillary histology) includes cancers that are iodine-refractory. In some embodiments, non-small cell lung cancer (NSCLC) includes cancers that progress during or after treatment with platinum-based chemotherapy and anti-PDx therapy for unresectable, locally advanced, or metastatic disease. In some embodiments, NSCLC with activating EGFR mutations (excluding exon 20 insertion mutations) or anaplastic lymphoma kinase (ALK) rearrangements must have progressed following treatment with platinum-based chemotherapy (unless ineligible for platinum therapy) plus available EGFR- or ALK-targeted therapy. In some embodiments, Merkel cell carcinoma includes metastatic Merkel cell carcinoma that is incurable by surgery or radiation.

[0283] In some embodiments of the methods of treating a disease or condition, the immune cells that exhibit differential proliferation include one or more of naive T cells, memory T cells, CD8+ T cells, NK cells, or natural killer T cells. In some embodiments of the methods of treating a disease or condition, the undesirable effects caused by IL-2 include one or more of activation of regulatory T cells, apoptosis of CD25+ T effector cells, IL-2-induced pulmonary edema, IL-2-induced pneumonia, or IL-2-induced vascular leakage. In some embodiments of the methods of treating a disease or condition, the anti-IL-2 antibodies disclosed herein inhibit binding of IL-2 to CD25.

[0284] In some embodiments, the treatment of cancer includes maintenance treatment. In some embodiments, the maintenance treatment is administered to maintain the absence of cancer or tumor. In some embodiments, the maintenance treatment is administered to maintain the absence of metastasis of the cancer or tumor. In some embodiments, the maintenance treatment is administered to inhibit metastasis of the cancer or tumor. In some embodiments, the maintenance treatment is administered to maintain the absence of growth of the cancer or tumor. In some embodiments, the maintenance treatment is administered to inhibit growth of the cancer or tumor.

[0285] In some embodiments, treating a solid cancer in a subject is reducing the size of the tumor, inhibiting or reducing the growth of the tumor, or inhibiting or reducing metastasis of said tumor, or any combination thereof.

[0286] In some embodiments, treating cancer includes preventative treatment, including, but not limited to, subjects who carry one or more genetic markers that are at increased risk of developing cancer. In some embodiments, the genetic markers include mutations in the BRCA1 gene.

[0287] Some embodiments of methods for promoting differential proliferation of immune cells in a subject include preparing and administering a composition comprising an anti-IL-2 antibody disclosed herein.

[0288] Some embodiments of the method for promoting differential proliferation of immune cells in a subject include preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody disclosed herein, wherein the administration of the combination of the anti-IL-2 antibody and the IL-2, or the composition thereof, is simultaneous. Some embodiments of the method for promoting differential proliferation of immune cells in a subject include preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody, wherein the administration of the anti-IL-2 antibody and the IL-2, or the composition thereof, comprises administering the anti-IL-2 antibody or the composition thereof before administering the IL-2 or the composition thereof. Some embodiments of the method for promoting differential proliferation of immune cells in a subject include preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody, wherein the administration of the anti-IL-2 antibody and the IL-2, or the composition thereof, comprises administering the anti-IL-2 antibody or the composition thereof after administering the IL-2 or the composition thereof.

[0289] In some embodiments, the present disclosure provides a method of treating a subject having a disease or condition through the induction of differential proliferation of immune cells. In one embodiment, the disease can be a viral infection, a bacterial infection, or cancer. In one embodiment, the condition can be IL-2-induced pulmonary edema or IL-2-induced vascular leakage. The method comprises: (a) preparing a composition comprising an anti-IL-2 antibody disclosed herein; (b) administering the composition from (a) to the subject, thereby treating the subject through differential proliferation of immune cells in the subject; In certain embodiments, any of the modified anti-IL-2 antibodies disclosed herein may be used in the described methods of treatment.

[0290] In some embodiments, the present disclosure provides a method of treating a subject having a disease or condition through the induction of differential proliferation of immune cells. In one embodiment, the disease can be a viral infection, a bacterial infection, or cancer. In one embodiment, the condition can be IL-2-induced pulmonary edema or IL-2-induced vascular leakage. The method comprises: (a) preparing a composition comprising IL-2 and an anti-IL-2 antibody disclosed herein; (b) administering the composition from (a) to the subject, thereby treating the subject through differential proliferation of immune cells in the subject; In addition to promoting the expansion of a subset of immune effector cells, the antibody / IL-2 complex will also reduce undesirable effects caused by IL-2 (e.g., IL-2-induced pulmonary edema or IL-2-induced vascular leakage). In one embodiment, the subject can be an animal or a human. In certain embodiments, any of the modified anti-IL-2 antibodies disclosed herein can be used in the described methods of treatment.

[0291] In one embodiment, the disclosure provides a method of treating a disease or condition in a subject (e.g., an animal or a human) comprising administering to the subject a composition comprising an anti-IL-2 antibody, wherein the antibody promotes the proliferation of a subset of immune cells and reduces undesirable effects caused by IL-2, thereby treating the disease or condition in the subject. Some embodiments of the method of treating a disease or condition in a subject comprise preparing and administering a composition comprising an anti-IL-2 antibody disclosed herein, and administering a composition comprising the anti-IL-2 antibody. In one embodiment, the composition comprises IL-2 and an anti-IL-2 antibody disclosed herein, or the composition comprises an anti-IL-2 antibody complexed to IL-2.

[0292] In some embodiments of the methods for treating a disease or condition in a subject, comprising preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody disclosed herein, the administration of the combination of the anti-IL-2 antibody and IL-2, or the composition thereof, is simultaneous. In some embodiments of the methods for treating a disease or condition in a subject, comprising preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody, the administration of the anti-IL-2 antibody and IL-2, or the composition thereof, comprises administration of the anti-IL-2 antibody or the composition thereof before the IL-2 or the composition thereof. In some embodiments of the methods for treating a disease or condition in a subject, comprising preparing and administering a composition comprising IL-2 and an anti-IL-2 antibody, the administration of the anti-IL-2 antibody and IL-2, or the composition thereof, comprises administration of the anti-IL-2 antibody or the composition thereof after administration of the IL-2 or the composition thereof.

[0293] In one embodiment, the method of treatment may be effective in treating conditions such as IL-2-induced pulmonary edema or IL-2-induced vascular leakage, hi another embodiment, the method of treatment may be effective in treating pulmonary edema (mild or chronic) resulting from viral or bacterial infection.

[0294] In one embodiment, the disease can be a viral infection, a bacterial infection, cancer, an autoimmune disease or immune disorder. In one embodiment, the disease can be an upper respiratory tract viral infection, an early pulmonary infection, or a late pulmonary infection. Several diseases and cancers are known to be caused by viruses. Examples of disease-causing viruses include, but are not limited to, norovirus, rotavirus, hepatitis A, B, C, D, or E virus, rabies virus, West Nile virus, enterovirus, echovirus, coxsackievirus, herpes simplex virus (HSV), HSV-2, varicella-zoster virus, mosquito-borne virus, arbovirus, St. Louis encephalitis virus, California encephalitis virus, lymphocytic choriomeningitis virus, human immunodeficiency virus (HIV), poliovirus, Zika virus, rubella virus, cytomegalovirus, human papillomavirus (HPV), enterovirus D68, severe acute respiratory syndrome (SARS) coronavirus, Middle East respiratory syndrome coronavirus, SARS coronavirus 2, Epstein-Barr virus, influenza virus, respiratory syncytial virus, polyomavirus (e.g., JC virus, BK virus), Ebola virus, dengue virus, or any combination thereof. In one embodiment, the viral infection is caused by SARS CoV-2. In another embodiment, the cancer may be, but is not limited to, melanoma or renal cell carcinoma.

[0295] In one embodiment, the immune cells expanded by treatment with an anti-IL-2 antibody include naive T cells, memory T cells, CD8 + These include one or more of T cells, NK cells, and natural killer T cells. In one embodiment, treatment with an anti-IL-2 antibody will reduce one or more undesirable effects caused by IL-2, such as activation of regulatory T cells, apoptosis of CD25+ T effector cells, pulmonary edema, pneumonia, and IL-2-induced vascular leakage.

[0296] In one embodiment, the anti-IL-2 antibody administered in the above methods is an altered or modified anti-IL-2 antibody capable of inhibiting binding of IL-2 to CD25. In some embodiments, the altered or modified anti-IL-2 antibody comprises a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, the altered or modified anti-IL-2 antibody comprises a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, the altered or modified anti-IL-2 antibody comprises heavy and light chain variable regions having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37.

[0297] In another embodiment, the altered or modified anti-IL-2 antibody comprises a heavy chain variable region having complementarity determining regions (CDRs) 1, 2, and 3. In one embodiment, heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38-40, respectively; 44-46, respectively; 50-52, respectively; 56-58, respectively; or 62-64, respectively.

[0298] In another embodiment, the altered or modified anti-IL-2 antibody comprises a light chain variable region having complementarity determining regions (CDRs) 1, 2, and 3. In one embodiment, light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41-43, respectively; 47-49, respectively; 53-55, respectively; 59-61, respectively; or 65-67, respectively.

[0299] In some embodiments, the modified anti-IL-2 antibody can be IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, or F(ab')2. The IgG can be of the IgG1, IgG2, IgG3, or IgG4 subclass. In some embodiments, the modified antibody can be part of a minibody, diabody, or triabody antibody.

[0300] In some embodiments, polynucleotide sequences encoding modified anti-IL-2 antibodies are used in methods of treating a subject having a disease or condition described herein, wherein the polynucleotide encodes an antibody comprising a heavy chain variable region having the amino acid sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments, polynucleotide sequences encoding modified anti-IL-2 antibodies are used in methods of treating a subject having a disease or condition described herein, wherein the polynucleotide encodes an antibody comprising a light chain variable region having the amino acid sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments, a polynucleotide sequence encoding a modified anti-IL-2 antibody is used in a method of treating a subject having a disease or condition described herein, wherein the polynucleotide encodes an antibody comprising a heavy chain variable region and a light chain variable region having the amino acid sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37.

[0301] In some embodiments of methods of using polynucleotides to treat the above diseases or conditions, the polynucleotide encodes a modified anti-IL-2 antibody that can be an IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, or F(ab')2. The IgG can be of the IgG1, IgG2, IgG3, or IgG4 subclass. In some embodiments, the polynucleotide encodes a modified antibody that is part of a minibody, diabody, or triabody antibody.

[0302] In some embodiments, a polynucleotide sequence encoding a modified anti-IL-2 antibody is used in a method of treating a subject having a disease or condition described herein, wherein the polynucleotide sequence comprises one of SEQ ID NOs: 1, 2, 3, 4, 5, 31, 32, 33, 34, or 35.

[0303] In some embodiments of the methods of treating a disease or condition described herein, the immune effector cells activated by the treatment are CD8+ cells or NK cells. In one embodiment, the anti-IL-2 antibody disclosed herein, or a conjugate of IL-2 and an anti-IL-2 antibody disclosed herein, activates MPCD8 + It shows a significant effect in inducing proliferation of CD4 cells and NK cells, but + The effect on Tregs was much smaller. In one particular embodiment, CD4 + There is no effect on Tregs.

[0304] In certain embodiments, the methods of use of the anti-IL-2 antibodies disclosed herein provide a pro-stimulatory effect. Those skilled in the art will appreciate that the use of the anti-IL-2 antibodies described and exemplified herein, e.g., Example 1, clearly demonstrates a pro-stimulatory effect, as opposed to an anti-stimulatory or pro-regulatory effect.

[0305] In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36 provides a stimulatory immune effect in a subject in need thereof. In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37 provides a stimulatory immune effect in a subject in need thereof. In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37 provides a stimulatory immune effect in a subject in need thereof. In some embodiments, the use comprises an anti-IL-2 antibody. In some embodiments, the use comprises an anti-IL-2 antibody and IL-2. In some embodiments, the use comprises a complex of an anti-IL-2 antibody and IL-2.

[0306] In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36 provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory or pro-regulatory effect. In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37 provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory or pro-regulatory effect. In some embodiments, the use of an engineered or modified anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region having one of the sequences of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37 provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory effect or a pro-regulatory effect. In some embodiments, the use comprises an anti-IL-2 antibody. In some embodiments, the use comprises an anti-IL-2 antibody and IL-2. In some embodiments, the use comprises a complex of an anti-IL-2 antibody and IL-2.

[0307] In some embodiments, the use of an anti-IL-2 antibody comprising a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 38-40, respectively; SEQ ID NOs: 44-46, respectively; SEQ ID NOs: 50-52, respectively; SEQ ID NOs: 56-58, respectively; or SEQ ID NOs: 62-64, respectively, provides a stimulatory immune effect in a subject in need thereof. In some embodiments, the use of an anti-IL-2 antibody comprising a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 41-43, respectively; SEQ ID NOs: 47-49, respectively; SEQ ID NOs: 53-55, respectively; SEQ ID NOs: 59-61, respectively; or SEQ ID NOs: 65-67, respectively, provides a stimulatory immune effect in a subject in need thereof. Variable regionThe use of an anti-IL-2 antibody comprising: a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; SEQ ID NOs: 44 to 46, respectively; SEQ ID NOs: 50 to 52, respectively; SEQ ID NOs: 56 to 58, respectively; or SEQ ID NOs: 62 to 64, respectively; and a light chain variable region comprising light chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; SEQ ID NOs: 47 to 49, respectively; SEQ ID NOs: 53 to 55, respectively; SEQ ID NOs: 59 to 61, respectively; or SEQ ID NOs: 65 to 67, respectively. Variable region Use of an anti-IL-2 antibody, including: provides a stimulatory immune effect in a subject in need thereof. In some embodiments, the use comprises an anti-IL-2 antibody. In some embodiments, the use comprises an anti-IL-2 antibody and IL-2. In some embodiments, the use comprises a complex of an anti-IL-2 antibody and IL-2.

[0308] In some embodiments, the use of an anti-IL-2 antibody comprising a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 38-40, respectively; SEQ ID NOs: 44-46, respectively; SEQ ID NOs: 50-52, respectively; SEQ ID NOs: 56-58, respectively; or SEQ ID NOs: 62-64, respectively, provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory effect or a pro-regulatory effect. In some embodiments, the use of an anti-IL-2 antibody comprising a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 41-43, respectively; SEQ ID NOs: 47-49, respectively; SEQ ID NOs: 53-55, respectively; SEQ ID NOs: 59-61, respectively; or SEQ ID NOs: 65-67, respectively, provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory effect or a pro-regulatory effect. Variable regionThe use of anti-IL-2 antibodies comprising: a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 38-40, respectively; SEQ ID NOs: 44-46, respectively; SEQ ID NOs: 50-52, respectively; SEQ ID NOs: 56-58, respectively; or SEQ ID NOs: 62-64, respectively; and a light chain variable region comprising light chain CDR1, CDR2, and CDR3 set forth in the amino acid sequences of SEQ ID NOs: 41-43, respectively; SEQ ID NOs: 47-49, respectively; SEQ ID NOs: 53-55, respectively; SEQ ID NOs: 59-61, respectively; or SEQ ID NOs: 65-67, respectively. Variable region The use of an anti-IL-2 antibody, including: provides a pro-stimulatory immune effect in a subject in need thereof, as opposed to an anti-stimulatory or pro-regulatory effect. In some embodiments, the use comprises an anti-IL-2 antibody. In some embodiments, the use comprises an anti-IL-2 antibody and IL-2. In some embodiments, the use comprises a complex of an anti-IL-2 antibody and IL-2.

[0309] Thus, the modified anti-IL-2 antibodies disclosed herein will be useful in modulating immune cell populations and inducing differential proliferation of certain immune effector cells in methods for treating diseases such as viral infections, bacterial infections, or cancer, or in methods for treating conditions such as IL-2-induced pulmonary edema or IL-2-induced vascular leakage.

[0310] Disclosed herein in some embodiments are methods of immunizing a subject, wherein the immunization comprises administering a vaccine comprising an adjuvant, wherein the adjuvant comprises an IL-2 antibody adjuvant. In some embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody and IL-2, or comprises an anti-IL-2 antibody complexed with IL-2. In some embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody and IL-2. In some embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody complexed with IL-2. In some embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody.

[0311] In some embodiments, the subject to be immunized is a mammalian subject. In some embodiments, the subject to be immunized is a human. In some embodiments, the subject to be immunized has a weakened immune system.

[0312] In some embodiments of the immunization methods, the anti-IL-2 antibody comprises a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36. In some embodiments of the immunization methods, the anti-IL-2 antibody comprises a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37. In some embodiments of the immunization methods, the anti-IL-2 antibody comprises an anti-IL-2 antibody comprising heavy and light chain variable regions having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37.

[0313] In some embodiments of the immunization method, the anti-IL-2 antibody includes an anti-IL-2 antibody comprising a heavy chain variable region including complementarity determining regions (CDRs) 1, CDR2, and CDR3, wherein the CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, 44 to 46, 50 to 52, 56 to 58, or 62 to 64, respectively. In some embodiments of the immunization method, the anti-IL-2 antibody includes an anti-IL-2 antibody comprising a light chain variable region including complementarity determining regions (CDRs) 1, CDR2, and CDR3, wherein the CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, 47 to 49, 53 to 55, 59 to 61, or 65 to 67, respectively. In some embodiments of the immunization method, the anti-IL-2 antibody comprises an anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region, each of the heavy chain variable region and the light chain variable region comprising complementarity determining region (CDR) 1, CDR2, and CDR3, wherein the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; SEQ ID NOs: 44 to 46, respectively; SEQ ID NOs: 50 to 52, respectively; SEQ ID NOs: 56 to 58, respectively; or SEQ ID NOs: 62 to 64, respectively; and the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; SEQ ID NOs: 47 to 49, respectively; SEQ ID NOs: 53 to 55, respectively; SEQ ID NOs: 59 to 61, respectively; or SEQ ID NOs: 65 to 67, respectively.

[0314] In some embodiments of the methods of immunizing a subject, immunization comprises administering a vaccine comprising an adjuvant, wherein the adjuvant comprises an IL-2 antibody adjuvant, and wherein the anti-IL-2 antibody comprises an anti-IL-2 antibody disclosed herein. In certain embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody and IL-2, or comprises an anti-IL-2 antibody complexed with IL-2. In some embodiments of the methods of immunizing a subject, the subject has a weakened immune system.

[0315] In some embodiments, subjects for immunization with a vaccine comprising an IL-2 antibody adjuvant include subjects suffering from a condition comprising a genetic predisposition that increases the subject's likelihood of cancer. In some embodiments, the genetic predisposition comprises an alteration in the expression or activity of a gene product. In some embodiments, the genetic predisposition that increases the likelihood of cancer comprises a mutation in a tumor suppressor gene or a mismatch repair (MMR) gene, or a combination thereof. Many hereditary cancers are known in the art, and non-limiting examples include, but are not limited to, hereditary breast and ovarian cancer (HBOC) syndrome, Lynch syndrome (hereditary nonpolyposis colorectal cancer), and Li-Fraumeni syndrome.

[0316] In some embodiments, a subject being treated by the methods disclosed herein for treating a disease or condition is further treated with one or more immune checkpoint inhibitors that target one or more immune checkpoint inhibitors. In some embodiments, the subject is treated with the immune checkpoint inhibitors simultaneously with, prior to, or following treatment with the anti-IL-2 antibody. In some embodiments of the methods of treatment disclosed herein, the immune checkpoints include PD-1, PD-L1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, or a combination thereof.

[0317] As discussed above, in some embodiments, the therapeutic methods disclosed herein further comprise an additional active agent, including a checkpoint inhibitor. One of skill in the art will appreciate that combination therapies, including anti-IL-2 antibody therapy with or without IL-2, and further comprising a checkpoint inhibitor, may utilize any of the therapeutic compositions or formulations comprising an anti-IL-2 antibody + / - IL-2 and a checkpoint inhibitor provided herein. In some embodiments, at least two checkpoint inhibitors are used in the combination therapy.

[0318] Embodiments of the present application include:

[0319] An isolated anti-IL-2 antibody comprising a heavy chain variable region comprising the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36.

[0320] Antibodies, including antibodies, including IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, F(ab')2, minibody, diabody, or triabody antibodies.

[0321] (a) IgG1, IgG2, IgG3, or IgG4; (b) a heavy chain containing a mutation that reduces binding to an Fcγ receptor (FcγR); or (c) lambda or kappa light chain; or (d) Any combination of (a) to (c) Including, IgG.

[0322] A composition comprising an isolated anti-IL-2 antibody and a pharmaceutically acceptable carrier.

[0323] An isolated anti-IL-2 antibody comprising a light chain variable region comprising the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37.

[0324] An isolated anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region having one of the sequences of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37.

[0325] An isolated anti-IL-2 antibody comprising a heavy chain variable region having complementarity determining region 1 (CDR1), CDR2 and CDR3, wherein the CDR1, CDR2 and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; SEQ ID NOs: 44 to 46, respectively; SEQ ID NOs: 50 to 52, respectively; SEQ ID NOs: 56 to 58, respectively; or SEQ ID NOs: 62 to 64, respectively.

[0326] An isolated anti-IL-2 antibody comprising a light chain variable region having complementarity determining region 1 (CDR1), CDR2 and CDR3, wherein the CDR1, CDR2 and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; SEQ ID NOs: 47 to 49, respectively; SEQ ID NOs: 53 to 55, respectively; SEQ ID NOs: 59 to 61, respectively; or SEQ ID NOs: 65 to 67, respectively.

[0327] an isolated anti-IL-2 antibody comprising: a heavy chain variable region having complementarity determining region 1 (CDR1), CDR2, and CDR3, wherein the CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, 44 to 46, 50 to 52, 56 to 58, or 62 to 64, respectively; and a light chain variable region having complementarity determining region 1 (CDR1), CDR2, and CDR3, wherein the CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, 47 to 49, 53 to 55, 59 to 61, or 65 to 67, respectively.

[0328] An isolated polynucleotide sequence encoding a heavy chain variable region of an anti-IL-2 antibody, wherein the heavy chain variable region comprises the amino acid sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36.

[0329] A vector comprising a polynucleotide sequence described herein. A host cell comprising a vector described herein.

[0330] An isolated polynucleotide sequence encoding a light chain variable region of an anti-IL-2 antibody, wherein the light chain variable region comprises the amino acid sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37.

[0331] An isolated polynucleotide sequence encoding a heavy chain variable region of an anti-IL-2 antibody comprising the amino acid sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36, and encoding a light chain variable region of an anti-IL-2 antibody comprising the amino acid sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37.

[0332] An isolated polynucleotide sequence encoding an scFv, the isolated polynucleotide sequence comprising one of SEQ ID NOs: 1, 2, 3, 4, 5, 31, 32, 33, 34, or 35.

[0333] A method for producing a heavy chain variable region of an anti-IL-2 antibody, the method comprising the step of culturing a host cell containing a vector disclosed herein under conditions conducive to expression of the vector in the host cell, thereby producing the heavy chain variable region of the anti-IL-2 antibody.

[0334] A method for producing a light chain variable region of an anti-IL-2 antibody, comprising culturing a host cell under conditions favoring expression of the vector in the host cell, thereby producing the light chain variable region of the anti-IL-2 antibody.

[0335] A method for producing an anti-IL-2 antibody comprising a heavy chain variable region and a light chain variable region of an anti-IL-2 antibody, the method comprising culturing a host cell under conditions conducive to expression of the vector in the host cell, thereby producing the anti-IL-2 antibody. Heavy chain variable region and producing a light chain variable region.

[0336] A method of promoting differential proliferation of immune cells in a subject, comprising administering a composition comprising an anti-IL-2 antibody, thereby promoting differential proliferation of immune cells in the subject. In some embodiments, the composition comprises an anti-IL-2 antibody and IL-2, or an anti-IL-2 antibody conjugated to IL-2.

[0337] A method for treating a subject with cancer through the induction of differential proliferation of immune cells, the method comprising administering a composition comprising an anti-IL-2 antibody, thereby treating the subject with cancer.

[0338] A method for treating a disease or condition in a subject, the method comprising administering to the subject a composition comprising an anti-IL-2 antibody, wherein the antibody promotes the proliferation of a subset of immune cells and reduces undesirable effects caused by IL-2, thereby treating the disease or condition in the subject.

[0339] In some embodiments, the disease comprises a viral infection, a bacterial infection, or cancer. In some embodiments, the viral infection is caused by SARS CoV-2, norovirus, rotavirus, hepatitis A, B, C, D, or E virus, rabies virus, West Nile virus, enterovirus, echovirus, coxsackievirus, herpes simplex virus (HSV), HSV-2, varicella-zoster virus, mosquito-borne virus, arbovirus, St. Louis encephalitis virus, California encephalitis virus, lymphocytic choriomeningitis virus, human immunodeficiency virus (HIV), poliovirus, Zika virus, rubella virus, cytomegalovirus, human papillomavirus (HPV), enterovirus D68, severe acute respiratory syndrome (SARS) coronavirus, Middle East respiratory syndrome coronavirus, Epstein-Barr virus, influenza virus, respiratory syncytial virus, polyomavirus including JC virus, BK virus, Ebola virus, dengue virus, or any combination thereof. In some embodiments, the condition involves a weakened immune system, and said treatment prophylactically boosts the immune system.

[0340] In some embodiments, the condition comprises IL-2 induced pulmonary edema.

[0341] In some embodiments of the methods disclosed herein, the immune cells comprise one or more of naive T cells, memory T cells, CD8+ T cells, NK cells, or natural killer T cells.

[0342] In some embodiments, the undesirable effects caused by IL-2 include one or more of activation of regulatory T cells, apoptosis of CD25+ T effector cells, IL-2-induced pulmonary edema, pneumonia, or IL-2-induced vascular leakage.

[0343] In some embodiments, the anti-IL-2 antibodies disclosed herein inhibit the binding of IL-2 to CD25.

[0344] A method of immunizing a subject, said immunization comprising administering a vaccine comprising an adjuvant, said adjuvant comprising an IL-2 antibody adjuvant.

[0345] In some embodiments, the IL-2 antibody adjuvant comprises an anti-IL-2 antibody and IL-2, or an anti-IL-2 antibody complexed with IL-2.

[0346] In some embodiments, the subject is an animal or a human. In some embodiments, the subject has a weakened immune system.

[0347] In some embodiments of the methods disclosed herein, the immune cells are CD8+ cells or NK cells.

[0348] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a heavy chain variable region having the sequence of one of SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, or 36.

[0349] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a light chain variable region having the sequence of one of SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, 25, 27, or 37.

[0350] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a heavy chain variable region and a light chain variable region having the sequence of one of SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 26 and 27; or SEQ ID NOs: 36 and 37.

[0351] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a heavy chain variable region including complementarity determining regions (CDRs) 1, 2, and 3, wherein the CDRs 1, 2, and 3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; 44 to 46, respectively; 50 to 52, respectively; 56 to 58, respectively; or 62 to 64, respectively.

[0352] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a light chain variable region including complementarity determining regions (CDRs) 1, 2, and 3, wherein the CDRs 1, 2, and 3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; 47 to 49, respectively; 53 to 55, respectively; 59 to 61, respectively; or 65 to 67, respectively.

[0353] In some embodiments of the methods disclosed herein, the anti-IL-2 antibody comprises a heavy chain variable region and a light chain variable region, each of which comprises a complementarity determining region (CDR) 1, CDR2, and CDR3, and the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequences of SEQ ID NOs: 38 to 40, respectively; SEQ ID NOs: 44 to 46, respectively; SEQ ID NOs: 50 to 52, respectively; SEQ ID NOs: 56 to 58, respectively; or SEQ ID NOs: 62 to 64, respectively; The light chain CDR1, CDR2 and CDR3 comprise the amino acid sequences of SEQ ID NOs: 41 to 43, respectively; SEQ ID NOs: 47 to 49, respectively; SEQ ID NOs: 53 to 55, respectively; SEQ ID NOs: 59 to 61, respectively; or SEQ ID NOs: 65 to 67, respectively.

[0354] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0355] Throughout this application, various embodiments of the present invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values ​​within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0356] Whenever a numerical range is indicated herein, this is intended to include any recited number (fractional or integer) within the indicated range. The phrases "ranging between" a first indicated number and a second indicated number and "ranging to" a first indicated number "from" a second indicated number are used interchangeably herein and are intended to include the first indicated number and the second indicated number and all fractional and integer numbers therebetween.

[0357] Those of ordinary skill in the art will understand that the term "about" can encompass deviations of 0.0001 to 5% from the indicated number or range of numbers. In some cases, the term "about" can encompass deviations of 1 to 10% from the indicated number or range of numbers. In some cases, the term "about" encompasses deviations of up to 25% from the indicated number or range of numbers. [Example]

[0358] Example 1 This example describes the generation of modified anti-IL-2 antibodies based on embodiments of the antibodies generated. The examples of generating modified anti-IL-2 antibodies are based on a subset of the antibodies disclosed herein. The description and results shown in Example 1 are exemplary and are not intended to limit the generation of modified anti-IL-2 antibodies disclosed throughout this application.

[0359] Library Design

[0360] A library was designed to introduce mutations into the JES6.1 sequence. The amino acid sequences of the heavy and light chain variable regions of JES6.1 are shown in SEQ ID NOs: 6 and 7, respectively. Briefly, three positions were varied with codons encoding all amino acids (codons NNS). The library design allowed for one mutation in both CDRs L3 and H3, as well as one mutation in one of the following CDRs: H1, H2, or L2. CDRs were defined by meeting the definitions of IMGT or ABR (Kunik et al., 2012). CDR residues that were conserved in the crystal structure of the mIL2-JES6.1 complex (PDB 4YQX) (based on a Blast search against the PDB database) or did not form specific interactions with mouse IL-2 (mIL-2) were excluded from mutation. The theoretical size of the library was 1.38E+7 mutants.

[0361] Library Selection

[0362] Screening and selection using yeast surface display

[0363] The yeast-displayed scFv library was grown in SDCAA selective medium and expression was induced with 2% w / v galactose overnight at 30°C according to established protocols. The library was incubated with 100 nM recombinant human IL-2 with a 6xHis tag (hIL-2-His) (Reprokine, Israel) in PBS 0.1% BSA for 1 hour, then washed three times with PBS 0.1% BSA and labeled with fluorescently labeled antibodies mouse anti-Myc-FITC (Santa Cruze, USA) and mouse monoclonal anti-His APC (Miltenyi Biotec, Germany, catalog 0020130-119-782). After labeling, the library was sorted for high-affinity binders to recombinant human IL-2 using a BioRad S3e fluorescence-activated cell sorter. Clones isolated from the final sort were sequenced by extracting plasmid DNA from yeast clones using a Zymoprep kit (Zymo Research, USA) and sequencing the DNA.

[0364] Koff selection

[0365] To select binders with improved off-rates, clones from round 2 selection were incubated with 10 nM 6xHis tag (hIL-2) for 15 min. Yeast were then washed three times with 1 ml PBS 0.1% BSA and incubated with 100 nM unlabeled IL-2 for 5 min, 4 h, 6 h, and 24 h. At the indicated time points, yeast were washed, labeled with Myc-FITC (Santa Cruze, USA) and monoclonal anti-His APC (Miltenyi Biotec, Germany, catalog 0020130-119-782), and screened with Se3 as described above.

[0366] IgG production

[0367] JES6.1 w.t. was purchased from Thermo Fisher (catalog: 16-7022-81). JES6.1.RMC was cloned as a rat Fv with a mouse IgG2a constant region and produced by GeneScript Antibody Production Services (Genscript NJ, USA). BDG17.0014 was cloned into a human IgG1 constant region and produced by GeneScript Antibody Production Services. The amino acid sequences of the heavy and light chain variable regions of JES6.1.RMC are shown in SEQ ID NO:8 and SEQ ID NO:9, respectively. All other antibodies were produced as described below.

[0368] Reformat

[0369] Selected scFv clones were reformatted to a human IgG1 format. The sequences of the light chain (LC) and heavy chain (HC) variable regions were optimized for mammalian codon usage and ordered as genblocks (GB) from IDT (Integrated DNA Technologies, Coralville, IA, USA). The GBs were cloned into pSF-CMV-HuIgG1_HC (HC plasmid) and pSF-CMV-HuLambda_LC (LC plasmid) using standard cloning techniques (Oxford Genetics, Oxford, UK). Where indicated, the variable heavy chain was cloned into pSF-CMV-HuIgG1_HC_LALA (HC plasmid), in which the DNA encoding L234 and L235 of the heavy chain was mutated to alanine codons (L234A, L235A).

[0370] IgG expression

[0371] Expi-CHO cells (Thermo Fisher Scientific, USA) were transfected with the LC and HC plasmids at a 2:1 ratio, and expression was performed according to the manufacturer's instructions. Briefly, 50 ml of Expi-CHO cells were cultured at 37°C, 120 rpm, and 8% CO2 at 6 × 10 6The cells were cultured to a density of 1000 cells / ml. Then, 50 μg of heavy and light chain expression plasmids were transfected into CHO cells at a 1:2 ratio. After transfection, booster enhancers and feed materials were added to the culture, and the growth conditions were changed to 32°C, 120 rpm, and 5% CO2. Ten days after transfection, cells were harvested. IgG was purified from the supernatant using Protein A beads (Tosoh Bioscience GmbH, Germany), followed by size-exclusion chromatography (SEC) purification on a Superdex 200 10 / 300 enrichment column (GE Healthcare, USA) using PBS as the mobile phase.

[0372] array

[0373] The DNA sequence encoding the scFv of clone 1 (17.021) is shown in SEQ ID NO: 1. The DNA sequence encoding the scFv of clone 2 (17.022) is shown in SEQ ID NO: 2. The DNA sequence encoding the scFv of clone 4 (17.023) is shown in SEQ ID NO: 3. The DNA sequence encoding the scFv of clone 5 (17.030) is shown in SEQ ID NO: 4. The DNA sequence encoding the scFv of clone 6 (17.035) is shown in SEQ ID NO: 5.

[0374] The original JES6_1 starting sequence and the amino acid sequences of the heavy and light chain variable regions of various anti-IL-2 clones are shown in the table below and in Figures 11 and 12.

[0375] [Table 2]

[0376] Measurement of IgG binding to human IL-2

[0377] SPR analysis was performed on a Biacore 200 (GE Healthcare, USA) on a CM5 chip (catalog number br10005-30, GE Healthcare, USA). The chip was crosslinked with a primary capture antibody against human IgG (catalog number br-1008-39, GE Healthcare, USA) or a primary capture antibody against mouse IgG (catalog number BR-1008-38, GE Healthcare, USA) to a target of 8000 RU. After crosslinking of the primary Ab, mouse and human test antibodies were immobilized on the primary Ab to a target of approximately 500 RU. JES6.1 was directly crosslinked onto the CM5 chip. Human IL-2 (catalog number 60568, Reprokine, Israel) analyte was run at concentrations ranging from 128 nM to 0.03 nM in a 2- or 3-fold dilution series, one concentration per cycle, in HEB-EP or PBS 0.05% Tween-20 (PBS-T) buffer. Mouse IL-2 (catalog: RKP04351, Reprokine, Israel) was flowed in HEB-EP or PBS-T buffer at concentrations ranging from 0.5 nM to 40 nM. At the end of each cycle, the analyte and tested antibody were stripped from the chip using 3 M MgCl2, and a new Ab to be tested was loaded onto the chip as described above. Where indicated, instead of stripping the antibody, kinetics were determined by injecting a range of analyte concentrations in one cycle using the single-cycle kinetic method. Binding kinetics were determined using a 1:1 binding model using Biacore T200 evaluation software.

[0378] Binding of IgG to cynomolgus monkey IL-2

[0379] SPR analysis was performed on a Biacore 200 (GE Healthcare, USA) on a CM5 chip (catalog number: br10005-30, GE Healthcare, USA). The chip was crosslinked with a primary capture Ab against human IgG (catalog number: br-1008-39, GE Healthcare, USA) up to 5000 RU of target, and cynomolgus IL-2 (cIL-2) was tested by the multi-cycle method under the same conditions as above.

[0380] SEC analysis.

[0381] To analyze IgG, 100 μg of sample was loaded onto a Superdex 200 10 / 300 gain column (GE Healthcare, USA) at a flow rate of 0.8 ml / min on a GE AKTA Explorer chromatography system (GE Healthcare, USA). Antibody retention time was monitored at 280 nm.

[0382] Test for specific binding to CD25 and CD122

[0383] To test specific binding to CD25, BDG17.023 was immobilized on a CM5 chip to a target RU of approximately 300 RU as described above. 50 nM IL-2 was then injected until BDG17.023 or a control antibody was saturated. The Ab-IL-2 complex was then washed with PBS-T buffer for 10 seconds, and 1000 nM CD25 was injected and the response monitored.

[0384] To test specific binding to CD122, BDG17.023 was immobilized on a CM5 chip to a target RU of approximately 300-500 RU, as described above. 50 nM hIL-2 was then injected until the BDG17.023 antibody was saturated with hIL-2. The Ab-IL-2 complex was then washed with PBS-T buffer for 10 seconds, and 1000 nM CD122 was injected and the response was monitored.

[0385] To test the specific binding of humanized antibody-IL-2 conjugates to CD122 and CD25, antibodies BDG17.038, BDG17.043, BDG17.053, BDG17.054, BDG17.067, and BDG17.069 (see Tables 6 and 7 in Example 2 for sequence information for these clones) were immobilized to a target RU of approximately 300 RU on a capture antibody bound CM5 chip channel as described above. 50 nM IL-2 was then injected until each antibody was saturated with hIL-2. The Ab-IL-2 conjugates were then washed with PBS-T buffer for 60 seconds, and 1000 nM CD25 was injected and the response monitored. Running buffer was then injected for 60 seconds to reach a steady baseline, after which 1000 nM CD122 was injected for 30 seconds at a flow rate of 30 μl / min. To test for CD122 binding, the same experiment was repeated in the reverse order, with CD122 injected first, followed by CD25.

[0386] DSF analysis of IgG Tm

[0387] To determine the T-start and Tm of the humanized anti-hIL-2 antibody, the antibody was diluted to 0.5 mg / ml in PBS and analyzed using a NanoDSF Prometheus NT.48 (Nanotemper, Germany) at a heating rate of 1 °C / min.

[0388] In vivo experiments

[0389] Treatment of mice with IL-2 / Ab complexes

[0390] Groups of six 7-8 week-old male C57BL / 6 mice were injected intraperitoneally (ip) daily for four consecutive days with BDG17.023 / hIL-2 or JES6.1 / mIL-2 immunoconjugates. PBS and free hIL-2 or mIL-2 served as controls. At the end of the fourth day, mice were sacrificed, and spleens were harvested and homogenized to a single cell suspension. Cells were filtered, centrifuged (400 g for 5 minutes), resuspended in 5 ml of PBS, and diluted to 5 × 10 6The final concentration of lymphocytes / ml was 1. Experiments were performed in accordance with the guidelines of the Israeli National Council on Animal Care and Use Committee (IACUC).

[0391] Groups of six 7-8 week-old male C57BL / 6 mice were injected intraperitoneally (IP) daily for four consecutive days with BDG17.038 / hIL-2, BDG17.043 / hIL-2, BDG17.054 / hIL-2, BDG17.038 / hIL-2, or isotype control / hIL-2 immunoconjugates. To form the complexes, 10 μg of antibody was preincubated with 0.5 μg of hIL-2 at 37°C for 30 minutes before injection. At the end of the fourth day, mice were sacrificed, and spleens were harvested and homogenized to a single cell suspension. Cells were filtered, centrifuged (400 g for 5 minutes), resuspended in 5 ml of PBS, and diluted to 5 × 10 6 The final concentration of lymphocytes / ml was 1. Experiments were performed in accordance with the guidelines of the Israeli National Council on Animal Care and Use Committee (IACUC).

[0392] B16F10 mouse melanoma tumor xenograft model

[0393] Female C57BL / 6 mice were injected with 2 × 10 5 B16-F10 tumor cells were inoculated subcutaneously. The tumor volume was approximately 30-50 mm. 3 Five days after inoculation, mice were randomized into experimental groups (n=10 / group) and received a single dose of 10 μg of anti-IL-2 antibody / 1 μg of the indicated antibody hIL-2 conjugate or PBS control intraperitoneally daily for four consecutive days. Mice were monitored for tumor volume growth, weight loss, and nonspecific clinical signs throughout the experiment.

[0394] Determination of immune cell populations by FACS

[0395] To identify immune cell ...

Claims

1. 1. A method of treating a solid cancer in a subject, comprising the step of administering to the subject a composition comprising an anti-IL-2 antibody, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3; The CDRs are (a) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, the LCDR2 comprises the amino acid sequence of DAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 54, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; having the amino acid sequence wherein the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the cancer in the subject.

2. The method of claim 1 , wherein multiple doses of the composition comprising an anti-IL2 antibody are administered.

3. and administering at least a single low dose of IL-2, wherein said low dose of IL-2 is about 15 x 10 per kg of said subject. ３ IU ~ 500 x 10 ３ 3. The method of claim 1 or claim 2, comprising IU.

4. 4. The method of claim 3, wherein said administering IL-2 comprises subcutaneous administration.

5. The method of claim 3 or 4, wherein the IL-2 is administered before, simultaneously with, or after administration of the anti-IL-2 antibody.

6. The method of any one of claims 3 to 5, wherein the IL-2 is administered as multiple doses.

7. 7. The method of claim 6, wherein the multiple doses of IL-2 are administered before, simultaneously with, or after administration of the anti-IL-2 antibody, or any combination thereof.

8. 8. The method of any one of claims 3 to 7, further comprising administering a checkpoint inhibitor.

9. 9. The method of claim 8, wherein the checkpoint inhibitor comprises PD-L1, PD-1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1.

10. 10. The method of any one of claims 1 to 9, wherein the solid cancer comprises melanoma, metastatic melanoma, primary and metastatic melanoma, renal cell carcinoma (RCC), non-small cell lung cancer (NSCLC), urothelial carcinoma, adrenocortical carcinoma, clear cell renal cell carcinoma (ccRCC), triple-negative breast cancer, head and neck squamous cell carcinoma (HNSCC), gastric or gastroesophageal cancer, esophageal squamous cell carcinoma, cutaneous squamous cell carcinoma (cSCC), pancreatic adenocarcinoma, cholangiocarcinoma, hepatocellular carcinoma (HCC), colorectal cancer (CRC), epithelial ovarian cancer, cervical cancer, endometrial cancer, thyroid cancer (follicular or papillary histology), or Merkel cell carcinoma.

11. 11. The method of any one of claims 1 to 10, comprising a second line treatment or a third line treatment, or a combination thereof.

12. 12. The method of any one of claims 1 to 11, wherein treating the subject reduces tumor size, inhibits or reduces tumor growth, or inhibits or reduces metastasis of the tumor, or any combination thereof.

13. The VH and VL are (a) the VH comprises the amino acid sequence of SEQ ID NO: 26 and the VL comprises the amino acid sequence of SEQ ID NO: 27; (b) the VH comprises the amino acid sequence of SEQ ID NO: 20 and the VL comprises the amino acid sequence of SEQ ID NO: 21; (c) the VH comprises the amino acid sequence of SEQ ID NO: 22 and the VL comprises the amino acid sequence of SEQ ID NO: 23; (d) the VH comprises the amino acid sequence of SEQ ID NO: 24 and the VL comprises the amino acid sequence of SEQ ID NO: 25; or (e) the VH comprises the amino acid sequence of SEQ ID NO: 36 and the VL comprises the amino acid sequence of SEQ ID NO: 37; 13. The method of any one of claims 1 to 12, wherein the amino acid sequence is:

14. The antibody is selected from the group consisting of IgG, IgA, IgM, IgE, IgD, Fv, scFv, Fab, and F(ab'). ２ 14. The method of any one of claims 1 to 13, comprising a fusion protein comprising a fusion protein, a minibody, a diabody, or a triabody.

15. 15. The method of any one of claims 1 to 14, wherein the antibody comprises a heavy chain comprising a mutation that reduces binding to an Fcγ receptor.

16. 16. The method of claim 15, wherein the mutations include L234A, L235A mutations.

17. When the HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, the LCDR2 comprises the amino acid sequence of DAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 67, the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 72 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 73; When the HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 49, the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 68 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO: 69; and When the HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 54, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 55, the amino acid sequence of the full-length heavy chain is set forth in SEQ ID NO: 70 and the amino acid sequence of the full-length light chain is set forth in SEQ ID NO:

71.

17. The method of any one of claims 1 to 16.

18. 1. A method of treating cancer in a subject, comprising administering to the subject an anti-IL-2 antibody and IL-2, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, the LCDR2 comprises the amino acid sequence of DAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 54, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; having the amino acid sequence wherein the IL-2 is administered by subcutaneous injection, the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, and the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, thereby treating the cancer in the subject.

19. 1. A method of treating a solid cancer in a subject, comprising the step of administering to the subject an anti-IL-2 antibody, IL-2, and a checkpoint inhibitor, wherein the IL-2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein the CDRs are: (a) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 64, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 65, the LCDR2 comprises the amino acid sequence of DAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 67; (b) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 38, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 39, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 40, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 43; (c) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 44, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 45, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 46, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 47, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 49; (d) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 50, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 51, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 52, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 53, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 54, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 55; or (e) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 56, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 57, the HCDR3 comprises the amino acid sequence of SEQ ID NO: 58, the LCDR1 comprises the amino acid sequence of SEQ ID NO: 59, the LCDR2 comprises the amino acid sequence of YAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 61; having the amino acid sequence the IL-2 is administered by subcutaneous injection, the dose of the anti-IL-2 antibody is about 0.5 mg to 12 mg per kg of the subject, the antibody promotes differential proliferation of immune cell subsets and reduces undesirable effects caused by IL-2, and the checkpoint inhibitor comprises PD-L1, PD-1, CTLA-4, TIGIT, TIM-3, B7-H3, CD73, LAG3, CD27, CD70, 4-1BB, GITR, OX40, SIRP-alpha (CD47), CD39, ILDR2, VISTA, BTLA, or VTCN-1, thereby treating the cancer in the subject.

20. 20. The method of claim 19, wherein the checkpoint inhibitor comprises a PD-L1 checkpoint inhibitor.

Images