Methods for Treating Tumors

Abstract

The present disclosure relates to a method of treating a tumor in a subject in need thereof, comprising administering an IL-27 inhibitor, wherein the subject is identified as having a tumor that (i) expresses WSX-1, (ii) contains one or more immune cells that express IL-27, or (iii) is both (i) and (ii).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 381,893, filed November 1, 2022, the entire contents of which are incorporated herein by reference.

[0002] Sequence Listing This application contains a computer-readable sequence listing entitled "01219-0017-00PCT_ST26", created on October 26, 2022, and having a size of 240,443 bytes, which is incorporated herein by reference.

[0003] Technical Field The present disclosure relates generally to methods of treating a tumor in a subject in need thereof, comprising administering a composition that modulates IL-27 signaling. More particularly, the present disclosure relates to methods of treating a tumor in a subject in need thereof, comprising administering an immunogenic composition (e.g., an antibody, antibody fragment, etc.) that binds to IL-27 and modulates IL-27 signaling. [Background technology]

[0004] In recent years, increasing evidence has suggested that the immune system serves as an important barrier against tumor formation and progression. The principle that natural T cells with antitumor potential or activity exist in cancer patients has rationalized the development of immunotherapeutic approaches in oncology. Immune cells, such as T cells, macrophages, and natural killer cells, exhibit antitumor activity and can effectively suppress the development and growth of malignant tumors. Tumor-specific or tumor-associated antigens can induce immune cells to recognize and eliminate malignant tumors (Chen & Mellman, (2013) Immunity 39(1):1-10). Despite the existence of tumor-specific immune responses, malignant tumors often evade or evade immune attack through various immunoregulatory mechanisms, resulting in uncontrolled tumor development and progression (Motz & Coukos, (2013) Immunity 39(1):61-730). Indeed, new hallmarks of cancer utilize these immunomodulatory mechanisms to neutralize antitumor immune responses, thereby allowing tumors to escape and evade immunological killing (Hanahan and Weinberg (2011) Cell 144(5):646-674).

[0005] IL-27 is a heterodimer composed of two subunits (EBI3 and IL-27p28). IL-27 is structurally related to both the IL-12 and IL-6 cytokine families. IL-27 binds to a heterodimeric receptor consisting of the IL-27Rα (WSX1) chain and the gp130 chain, and mediates signaling primarily through STAT1 and STAT3. Early reports characterized IL-27 as an immune-enhancing cytokine that supports CD4+ T cell proliferation, T helper (Th)1 cell differentiation, and IFN-γ production, often acting in concert with IL-12. Subsequent studies have revealed that IL-27 exhibits complex immunoregulatory functions, exerting either pro- or anti-inflammatory effects depending on the biological context and experimental model used. IL-27 promotes the expression of various immunoregulatory molecules in human cancer cells, which may support localized perturbation of immune responses in vivo (Fabbi et al., (2017) Mediators Inflamm 3958069. Published online 2017 Feb 1. doi:10.1155 / 2017 / 3958069 and references therein). Summary of the Invention [Problem to be solved by the invention]

[0006] Despite significant advances in the treatment and management of cancer, there remains a need for new and effective therapies for the treatment and management of cancer. [Means for solving the problem]

[0007] Some embodiments of the present disclosure relate to a method for treating a tumor in a subject, comprising administering an IL-27 inhibitor to the subject, wherein the tumor is identified as a WSX-1-positive tumor. Some embodiments of the present disclosure relate to a method for treating a tumor in a subject in need of treatment, comprising: (i) identifying a subject with a WSX-1-positive tumor; and (ii) administering an IL-27 inhibitor to the subject. In some embodiments, the WSX-1-positive tumor is identified by detecting WSX-1 expression in a tumor sample obtained from the subject.

[0008] Some embodiments of the present disclosure relate to a method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, comprising detecting WSX-1 expression in a tumor sample obtained from the subject. In some embodiments, the method further comprises administering an IL-27 inhibitor to the subject identified as having a WSX-1-positive tumor.

[0009] In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy. In some embodiments, the tumor sample obtained from the subject is a formalin-fixed, paraffin-embedded tumor sample. In some embodiments, the tumor sample obtained from the subject comprises tumor cells, tumor-infiltrating immune cells, or both.

[0010] In some embodiments, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of the cells in a tumor sample express WSX-1. In some embodiments, at least about 1% of the cells in a tumor sample express WSX-1.

[0011] In some embodiments, WSX-1 expression is detected using an immunohistochemistry (IHC) assay. In some embodiments, WSX-1 expression is detected using an automated IHC assay. In some embodiments, WSX-1 expression is scored using a tumor proportion score (TPS) and / or a combined positive score (CPS). In some embodiments, the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50%, or at least 60%. In some embodiments, WSX-1 expression is detected by contacting a tumor sample with an antibody or antigen-binding portion thereof that specifically binds to human WSX-1.

[0012] Some embodiments of the present disclosure relate to methods for treating a tumor in a subject, comprising administering to the subject an IL-27 inhibitor, wherein one or more immune cells in a tumor sample obtained from the subject express IL-27.

[0013] Some embodiments of the present disclosure relate to methods for treating a tumor in a subject in need thereof, the method comprising: (i) identifying a subject having a tumor in which one or more immune cells in a tumor sample obtained from the subject express IL-27; and (ii) administering to the subject an IL-27 inhibitor.

[0014] Some embodiments of the present disclosure relate to a method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, the method comprising detecting IL-27 expression in a tumor sample obtained from the subject. In some embodiments, the method further comprises administering an IL-27 inhibitor to a subject identified as having a tumor sample comprising one or more immune cells that express IL-27.

[0015] In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy.In some embodiments, the tumor sample obtained from the subject is a formalin-fixed paraffin-embedded tumor sample.In some embodiments, the tumor sample obtained from the subject comprises tumor cells, tumor-infiltrating immune cells, or both.In some embodiments, one or more immune cells in the tumor sample comprise macrophages.

[0016] In some embodiments, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 1% of immune cells in a tumor sample express IL-27.

[0017] In some embodiments, IL-27 expression is detected using an immunohistochemistry (IHC) assay. In some embodiments, IL-27 expression is detected using an automated IHC assay. In some embodiments, IL-27 expression is assessed using a tumor proportion score (TPS) and / or a combined positive score (CPS). In some embodiments, the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50%, or at least 60%. In some embodiments, IL-27 expression is detected by contacting a tumor sample with an antibody or antigen-binding portion thereof that specifically binds human IL-27.

[0018] In some embodiments, the IL-27 inhibitor reduces or blocks the interaction of IL-27 with WSX-1. In some embodiments, the IL-27 inhibitor comprises a polypeptide or a small molecule. In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27 (an "anti-IL-27 antibody").

[0019] In some embodiments, an IL-27 inhibitor increases the expression of GBP5 and IRF1. In some embodiments, an IL-27 inhibitor increases the expression of GBP5 and IRF1 in NK cells and / or CD8+ T cells.

[0020] In some embodiments, the anti-IL-27 antibody specifically binds to an epitope on human IL-27 that includes one or more amino acids of (i) amino acids 37 to 56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142 to 164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii). In some embodiments, the epitope comprises one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, or Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, the epitope comprises Asp146, Arg149, and / or Phe153 of SEQ ID NO: 2 (IL-27p28). In some embodiments, the epitope further comprises His150 and / or Leu156 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope further comprises Gln37, Leu38, Glu42, Leu142, and / or Glu164 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope further comprises Glu46, Val49, Ser50, and / or Leu162 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phel53, Leu156, Leu162, and Glu164 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope further comprises one or more amino acids of Leu53, Lys56, Asp143, Leu147, Arg152, Ala157, Gly159, Phe160, or Asn161 of SEQ ID NO: 2 (IL-27p28).In some embodiments, the epitope further comprises Leu53, Lys56, Asp143, Arg145, Leu147, Arg152, Ala157, Gly159, Phe160, Asn161, or Pro163 of SEQ ID NO: 2 (IL-27p28). In some embodiments, the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0021] In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3, a light chain CDR1, a light chain CDR2, and a light chain CDR3, wherein (i) the light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-C and the light chain CDR3 consists of N-XXXASAXXX-C; the heavy chain CDR2 consists of N-XXSSSXSYXYXXXXXXX-C; and the heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, where X is any amino acid.

[0022] In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR3 comprising the sequence set forth in SEQ ID NO: 121 or 124. In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR2 comprising the sequence set forth in SEQ ID NO: 120 or 123. In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the sequence set forth in SEQ ID NO: 119 or 122. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR3 comprising the sequence set forth in SEQ ID NO: 129 or 132. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR2 comprising the sequence set forth in SEQ ID NO: 128 or 131. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the sequence set forth in SEQ ID NO: 127 or 130.

[0023] In some embodiments, the antibody or antigen-binding portion thereof comprises (i) a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121; or (i) a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 122, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124.

[0024] In some embodiments, the antibody or antigen-binding portion thereof comprises (i) a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; or (ii) a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 132.

[0025] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121, a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129.

[0026] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 122, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 123, a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124, a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 132.

[0027] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 125. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 125. In some embodiments, the antibody or antigen-binding portion thereof is a light chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 133. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 125 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 133. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 135. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 139. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 135 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 139 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 137.

[0028] In some embodiments, the cancer is selected from lung cancer (e.g., non-small cell lung cancer), sarcoma, testicular cancer, ovarian cancer, pancreatic cancer, breast cancer (e.g., triple-negative breast cancer), melanoma, head and neck cancer (e.g., head and neck squamous cell carcinoma), colon cancer, bladder cancer, endometrial cancer, prostate cancer, thyroid cancer, hepatocellular carcinoma (HCC), gastric cancer, brain tumor, lymphoma (e.g., DL-BCL), leukemia (e.g., AML), kidney cancer (e.g., renal cell carcinoma (RCC), e.g., clear cell RCC and / or non-clear cell RCC), and any combination thereof.

[0029] In some embodiments, the method further comprises administering an additional therapeutic agent to the subject. In some embodiments, the additional therapeutic agent is administered before, after, or simultaneously with the antibody or antigen-binding portion thereof. In some embodiments, the additional therapeutic agent comprises chemotherapy, targeted anti-cancer therapy, oncolytic agent, cytotoxic agent, immune-based therapy, cytokine, surgery, radiation treatment, activator of costimulatory molecules, inhibitor of inhibitory molecules, vaccine, cellular immunotherapy, biologic agent, or a combination thereof.

[0030] In some embodiments, the additional therapeutic agents comprise a PD-1 antagonist, a PD-L1 inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor, a TIGIT inhibitor, a CD112R inhibitor, a TAM inhibitor, a STING agonist, a 4-1BB agonist, a multi-tyrosine kinase inhibitor (e.g., a VEGFR inhibitor), an anti-VEGF blocking antibody, a CTLA-4 antagonist, a HIF2 antagonist, a TGFb antagonist, an mTOR inhibitor, an adenosine pathway inhibitor (e.g., an anti-CD73 antibody, an anti-CD39 antibody, an anti-A2AR antibody, an anti-A2BR, or any combination thereof), an anti-CCR8 antibody, a cytokine-based regimen (e.g., IL-2 or IFN-a), a PARP inhibitor, or a combination thereof.

[0031] In some embodiments, the additional therapeutic agent comprises a PD-1 antagonist, hi some embodiments, the PD-1 antagonist is selected from the group consisting of PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, and AMP-224.

[0032] In some embodiments, the PD-L1 inhibitor is selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559.

[0033] In some embodiments, the additional therapeutic agent is sunitinib (SUTENT®), cabozantinib (CABOMETYX®), axitinib (INLYTA®), lenvatinib (LENVIMA®), everolimus (AFINITOR®), bevacizumab (AVASTIN®), epacadostat, NKTR-214 (CD-122 biased agonist), tivozanib (FOTIVDA®), abexinostat, ipilimumab ( YERVOY®), tremelimumab, pazopanib (VOTRIENT®), sorafenib (NEXAVAR®), temsirolimus (TORISEL®), ramucirumab (CYRAMZA®), niraparib, savolitinib, borolinib (X-82), regorafenib (STIVARGO®), donafenib (multikinase inhibitor), camrelizumab (SHR-1210), pexastimogene devasilepvec (pexastimogene devacirepvec (JX-594), ramucirumab (CYRAMZA®), apatinib (YN968D1), encapsulated doxorubicin (THERMODOX®), tivantinib (ARQ197), ADI-PEG 20, binimetinib, apatinib mesylate, nintedanib, lirilumab, nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFIMZI®), cemiplimab-rwlc (LIBTAYO®), tislelizumab, and spartalizumab.

[0034] In some embodiments, the additional therapeutic agent is a TIM-3 inhibitor. In some embodiments, the TIM-3 inhibitor is MGB453 or TSR-022.

[0035] In some embodiments, the additional therapeutic agent is a LAG-3 inhibitor. In some embodiments, the LAG-3 inhibitor is selected from the group consisting of LAG525, BMS-986016, and TSR-033.

[0036] In some embodiments, the additional therapeutic agent is a TIGIT inhibitor. In some embodiments, the additional therapeutic agent is a CD112R inhibitor. In some embodiments, the additional therapeutic agent is a TAM (Axl, Mer, Tyro) inhibitor. In some embodiments, the additional therapeutic agent is a 4-1BB agonist. In some embodiments, the additional therapeutic agent is a tyrosine kinase inhibitor (TKI).

[0037] Some embodiments of the present disclosure relate to kits that include: (i) an antibody or antigen-binding portion thereof that specifically binds to human WSX-1; (ii) an IL-27 inhibitor; and (iii) instructions for using (i) and (ii) in the methods disclosed herein.

[0038] Some embodiments of the present disclosure relate to kits that include: (i) an antibody or antigen-binding portion thereof that specifically binds to human IL-27; (ii) an IL-27 inhibitor; and (iii) instructions for using (i) and (ii) in the methods disclosed herein. [Brief explanation of the drawings]

[0039] [Figure 1-1]Figures 1A–1J present data showing WSX-1 mRNA expression (Figure 1A) and protein expression (Figures 1B–1J). Figure 1A is a boxplot showing WSX-1 mRNA expression in non-heme cell lines. Figures 1B–1F are IHC images of WSX-1 protein in FFPE tumor samples from lung adenocarcinoma (Figure 1B), lung squamous cell carcinoma (SCC), ovarian cancer, head and neck squamous cell carcinoma (HNSCC), and triple-negative breast cancer (TNBC). Figure 1G is a bar graph summarizing the percentage of cases that showed tumor cell staining for WSX-1 by IHC. Figures 1H–1I are example images of lung adenocarcinoma showing tumor cell staining for WSX-1 in both the primary tumor (Figure 1H) and synchronous lymph node metastasis (Figure 1I). FIG. 1J is a bar graph showing the percentage of lymph node metastatic NSCLC cases showing tumor cell staining for WSX-1 by IHC, grouped by WSX-1 status of the primary tumor. [Figure 1-2] Same as above. [Figure 1-3] Same as above. [Figure 1-4] Same as above. [Figure 1-5] Same as above. [Figure 2-1] Figures 2A–2F are example anti-IL-27 immunohistochemistry images of tumor samples from lung SCC (Figure 2A), clear cell renal cell carcinoma (ccRCC, Figure 2B), gastric cancer (Figure 2C), hepatocellular carcinoma (HCC, Figure 2D), HNSCC (Figure 2E), and lung adenocarcinoma (Figure 2F), showing positive cells within the tumor microenvironment (TME) morphologically consistent with tumor-associated macrophages (TAMs). Figure 2G is a graphical representation of the density (cells / mm) of IL-27+ cells in various cancer types. Figure 2H is an sc-RNA-seq analysis of the Immune Atlas on various cancer tumor cells for overall IL-27 expression across all cancer types studied. [Figure 2-2] Same as above. [Figure 2-3] Same as above. [Figure 3-1]Figures 3A-3C are graphical representations of the correlation between IL-27 and PD-L1 expression in NSCLC (Figure 3A), gastric cancer (Figure 3B), and HCC (Figure 3C). **p<0.01; ***p<0.001. Figures 3D-3I show examples of IHC for PD-L1 and IL-27 in gastric cancer samples with various CPS scores. The CPS score correlates with the density of IL-27+ cells (left and center), but a proportion of tumor samples with a CPS of less than 1 (PD-L1 negative) contain IL-27+ cells (right). All images are at 17x magnification. [Figure 3-2] Same as above. [Figure 3-3] Same as above. [Figure 3-4] Same as above. [Figure 4-1] Figures 4A-4I show example images of IHC for IL-27 (Figures 4A, 4D, and 4G), WSX-1 (Figures 4B, 4E, and 4H), and PD-L1 (Figures 4C, 4F, and 4I) performed on serial sections of normal human tonsil (Figures 4A-4C) and lung SCC (Figures 4D-4I). [Figure 4-2] Same as above. [Figure 5] Figure 5 shows example images of IHC for IL-27 performed on NSCLC tumor samples. Middle image: 0.9x magnification, inset: 12x magnification. [Figure 6] Figures 6A-6D show example images of WSX-1 IHC performed on NSCLC tumor samples. WSX-1 cellular staining is demonstrated in the peritumoral immune cell infiltrate (Figure 6A, 0.55x magnification), tertiary lymphoid structures (TLS; Figure 6B, 3.5x magnification), follicular dendritic cells (FDCs) within the germinal centers of TLS (Figure 6C, 12x magnification), and outside the TLS (Figure 6D, 20x magnification). [Figure 7-1] Figure 7A provides example images of IHC for IL-27 (left) and WSX-1 (right) performed on NSCLC tumor samples obtained as whole sections from individual lobectomy specimens. Figure 7B provides a graph of semiquantitative scoring of immune cells in tumor tissue areas. Bars represent the mean ± SEM of n=44 NSCLC lobectomy specimens. [Figure 7-2] Same as above. [Figure 8-1] Figures 8A-8B provide example images of IHC for IL-27 (left) and WSX-1 (right) on FFPE samples of NSCLC draining lymph nodes obtained from individual lobectomy specimens in lymph node metastases (Figure 8A; 10x magnification) and tumor-free draining lymph nodes (Figure 8B; 7x magnification). Figure 8C provides example images of IHC for IL-27 (left) and WSX-1 (right) in control lymph nodes. [Figure 8-2] Same as above. [Figure 8-3] Same as above. [Figure 9-1] Figures 9A-9B provide example images of IHC for IL-27 (Figure 9A, 20x magnification) and WSX-1 (Figure 9B, 20x magnification) in a subset of NSCLC samples. Figure 9C provides a graph showing the percentage of cases with positive staining in tumor cells. Bars represent the mean ± SEM of n=44 NSCLC lobectomy specimens. [Figure 9-2] Same as above. [Figure 10-1] Figure 10A provides example images of IHC for IL-27 (left), WSX-1 (center), and CD8 (right) in an NSCLC case (2.2x magnification). Figure 10B provides example images of IHC for IL-27 (left), PD-L1 (center), and CD8 (right) in an NSCLC case (4x magnification [top] and 10x magnification [bottom]). [Figure 10-2] Same as above. [Figure 11-1] Figures 11A-11B provide example images of IHC for IL-27 and PD-L1 in NSCLC cases. Figure 11A shows IHC in lung adenocarcinoma (4x magnification [top] and 20x magnification [bottom]). Figure 11B shows IHC in lung squamous cell carcinoma (6x magnification [top] and 18x magnification [bottom]). [Figure 11-2] Same as above. [Figure 12-1]Figures 12A-12D are graphical representations of the correlation between IL-27 and WSX-1 expression and PD-L1 expression in tumors classified using the tumor proportion score (TPS) system (Figures 12A, 12C) or the composite proportion score (CPS) system (Figures 12B, 12C). [Figure 12-2] Same as above. [Figure 12-3] Same as above. [Figure 12-4] Same as above. [Figure 13-1] Figures 13A-13B are graphical representations of the correlation between IL-27 and WSX-1 in patients who subsequently respond to immune checkpoint blockade (ICP) (Figure 13A), and the correlation between response to ICP and PD-L1 status (Figure 13B). Data points are mean ± SEM for n=24 NSCLC lobectomy specimens. Figures 13C-13D are graphical representations of the correlation between immune cell expression of IL-27 and WSX-1 in patients who received ICP as first-line therapy (Figure 13C) and in patients who received ICP as second-line or more advanced therapy (Figure 13D). TPS: tumor proportion score; CPS: composite proportion score. *: p<0.01 by Student's t-test. All other comparisons were not statistically significant. PD: progressive disease; SD: stable disease; PR: partial response; CR: complete response. The number of patients in each response category is indicated below the response label. [Figure 13-2] Same as above. [Figure 13-3] Same as above. [Figure 13-4] Same as above. [Figure 14-1]Figures 14A-14B are graphical representations of IHC scoring on archival specimens from NSCLC patients subsequently treated with SRF388 using TAM scoring (Figure 14A) or positive IL-27+ immune cells as a percentage of tumor area (Figure 14B). Squares indicate archival biopsy specimens; circles indicate archival resection specimens. Black circles / squares indicate partial responses to SRF388 monotherapy. Gray circles / squares indicate stable disease. White circles / squares indicate progressive disease. Figures 14C-14F provide IHC images of IL-27 in an archival lung resection specimen (Figure 14C, 20x magnification), including those adjacent to tertiary lymphoid structures (TLS) (Figure 14D, 15x magnification; asterisks indicate TLS) and residual lymph node tissue in Figure 14F (Figure 14E, 8x magnification, and 20x magnification). [Figure 14-2] Same as above. [Figure 14-3] Same as above. [Figure 15-1] Figure 15A is a graphical representation of IHC scoring of an archived specimen from a hepatocellular carcinoma (HCC) patient subsequently treated with SRF388 in combination with atezolizumab and bevacizumab using TAM scoring. Figures 15B-15F provide example images of IHC for IL-27 in a patient who responded to SRF388 combination therapy (Figures 15B, 15E, 15F, 20x magnification). Figures 15C and 15D focus on the IL-27+ macrophages in Figure 15B. [Figure 15-2] Same as above. [Figure 15-3] Same as above. [Figure 15-4] Same as above. [Figure 15-5] Same as above. [Figure 16] Figure 16A provides an experimental overview of a single-cell RNA sequencing experiment comparing IL-27 to various interferons. Seurat-based clustering was used to identify cell subsets assigned based on differential gene expression. Figure 16B provides a UMAP view of aggregated data from all conditions identifying distinct cell populations. [Figure 17-1]Figure 17A provides a schematic of IL-27 and type 2 / IFNG and type 1 / IFNB1 interferon transcript expression in activated PBMCs based on scRNA-seq. Figure 17B provides a heatmap projection of IL-27, IFNG, and IFNB1 transcript expression by various cell types in activated PBMCs. Figure 17C provides a dot plot of IL-27 expression in activated PBMCs after stimulation with various cytokines. [Figure 17-2] Same as above. [Figure 17-3] Same as above. [Figure 18-1] Figures 18A-C provide data showing that IL-27 and interferons upregulate the expression of several canonical interferon-stimulated genes. Figure 18A provides a volcano plot showing genes up- or down-regulated in PBMCs by the indicated cytokines. Figure 18B provides a Venn diagram showing the overlap in the top 100 genes up-regulated by IFNB1, IFNG, and IL-27. Figure 18C provides a heatmap representation of the 17 common genes among the top 100 genes up-regulated by IFNB1, IFNG, and IL-27. [Figure 18-2] Same as above. [Figure 18-3] Same as above. [Figure 19-1]Figure 19A provides a heatmap representation of immune checkpoint receptor gene expression showing similar upregulation of immunoregulatory receptors TIM-3 (HAVCR2), PD-L1 (CD274), TIGIT, and LAG3 by IL-27 or type 1 or type 2 IFN in CD4+ T cells, CD8+ T cells, and CD14hi monocytes. Figure 19B provides a heatmap representation of cytokine gene expression showing differential changes in transcripts for GM-CSF (CSF2), IFNG, IL17A, IL17F, and IL10 by IL-27 or type 1 or type 2 interferons in CD4+ T cells and proliferating T cells (defined by MK167+ or having G2M or S phase Seurat assignment, but with similar IFNG upregulation in CD8+ T cells by all three cytokines). [Figure 19-2] Same as above. [Figure 20-1] Figure 20A provides a UMAP display from cells of activated PBMCs cultured with various cytokines (Ctrl, IFNB1, IFNG, IL-27). Darker colors represent higher expression of the composite IFN gene signature, while lighter colors indicate lower expression. Figure 20B provides a heatmap display of cytokine receptor expression across various immune cell populations. Note the low expression of type 2 interferon receptor (IFNR) on CD4+ / CD8+ T cells and NK cells. [Figure 20-2] Same as above. [Figure 21-1] Figure 21A provides flow cytometry data measuring STAT1 phosphorylation in human PBMCs after 30 minutes of incubation with the indicated cytokines. Quantitative statistical analysis of cytokine-induced pSTAT1 (geometric mean fluorescence intensity, gMFI) in various cell types from PBMCs of multiple healthy donors (n=12). Figure 21B provides graphical data showing that type 1 and IL-27 robustly induce STAT1 phosphorylation in T cells, NK cells, and monocytes, whereas type 2 (IFNG)-mediated STAT1 phosphorylation occurs primarily in monocytes (representative data from several experiments shown). [Figure 21-2] Same as above. [Figure 22-1] Figure 22A provides cytokine gene expression signatures displayed as composite signatures (dark = high, light = low) and highlights the CD8+ and NK cell populations used for volcano plot analysis. Figure 22B provides a volcano plot analysis of differential gene expression by IL-27 compared to IFNB1 or IFNγ in NK cells and CD8+ T cells. Figure 22C is a heatmap representation of genes preferentially regulated by IL-27, type 1, or type 2 IFNγ in various immune cell populations. Note that T cells include both CD8+ and CD4+ T cells. [Figure 22-2] Same as above. [Figure 22-3] Same as above. [Figure 23-1] Figure 23A provides a violin plot of GBP5 transcript expression in T cells and NK cells after stimulation with IL-27 or type 1 or type 2 IFN. Figure 23B provides flow cytometry data of intracellular GBP5 expression in various cell types after 24 hours of stimulation of PBMCs with the indicated cytokines (representative data from several experiments are shown). Figure 23C provides an example immunohistochemistry (IHC) image from a treatment-naive NSCLC patient sample, showing GBP5+ T cell-rich areas within the TME colocalizing with IL-27+ macrophages and PD-L1+ immune cells. The T cell-rich areas are highlighted by CD4 IHC. [Figure 23-2] Same as above. [Figure 23-3] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0040] Some embodiments of the present disclosure relate to methods for treating a tumor in a subject, comprising administering an IL-27 inhibitor to the subject, wherein the tumor is identified as (i) a WSX-1-positive tumor or (ii) containing one or more immune cells within the tumor that express IL-27. Some embodiments of the present disclosure relate to methods for treating a tumor in a subject in need of treatment, comprising (i) identifying a subject having (a) a WSX-1-positive tumor, or (b) a tumor containing one or more immune cells within the tumor that express IL-27, and (ii) administering an IL-27 inhibitor to the subject. Some embodiments of the present disclosure relate to methods for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, comprising detecting (i) WSX-1 expression or (ii) IL-27 expression in a tumor sample obtained from the subject.

[0041] I. Definition Terms used in the claims and specification are defined as follows, unless otherwise specified.

[0042] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly indicates otherwise.

[0043] As used herein, "about" will be understood by one of ordinary skill in the art and will vary to some extent depending on the context in which it is used. If there are uses of the term that are not clear to a person of ordinary skill in the art given the context in which the term is used, "about" will mean up to ±10% of the particular value.

[0044] As used herein, the term "agonist" refers to any molecule that partially or completely promotes, induces, enhances, and / or activates the biological activity of a native polypeptide disclosed herein. Suitable agonist molecules include, in particular, agonist antibodies or antibody fragments, fragments or amino acid sequence variants of native polypeptides, peptides, or proteins. In some embodiments, activation in the presence of an agonist is observed in a dose-dependent manner. In some embodiments, the measured signal (e.g., biological activity) is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% higher than the signal measured using a negative control under comparable conditions. Also disclosed herein are methods for identifying agonists suitable for use in the methods of the present disclosure. For example, these methods include, but are not limited to, binding assays such as enzyme-linked immunosorbent assay (ELISA), FORTE BIO® system, and radioimmunoassay (RIA). These assays determine the ability of an agonist to bind to a polypeptide of interest (e.g., a receptor or ligand) and thereby promote, increase, or activate the activity of the polypeptide. The effectiveness of an agonist can also be determined using a functional assay, such as the ability of the agonist to activate or promote the function of a polypeptide. For example, a functional assay can include contacting a polypeptide with a candidate agonist molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide. The potency of an agonist is typically measured using an EC 50 The EC value is defined as the concentration required to activate 50% of the agonist response. 50 The lower the value, the more potent the agonist and the lower the concentration required to activate a maximal biological response.

[0045] The term "amelioration" refers to any therapeutically beneficial result in the treatment of a disease state, such as cancer, including prevention, reduction in severity or progression, remission, or cure thereof.

[0046] As used herein, the term "amino acid" refers to natural and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to natural amino acids. Natural amino acids are those encoded by the genetic code, as well as amino acids that are subsequently modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as natural amino acids, i.e., carbon atoms bonded to hydrogen, carboxyl groups, amino groups, and R groups, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as natural amino acids. Amino acid mimetics refer to compounds that have a structure different from the general chemical structure of amino acids but function similarly to natural amino acids.

[0047] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides may also be referred to by their commonly accepted single-letter codes.

[0048] As used herein, "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a predetermined amino acid sequence (the amino acid sequence of the starting polypeptide) with a second, different, "substitution" amino acid residue. "Amino acid insertion" refers to the incorporation of at least one additional amino acid into a predetermined amino acid sequence. Insertions typically consist of the insertion of one or two amino acid residues, although larger "peptide insertions," e.g., of about 3 to about 5, or up to about 10, 15, or 20 amino acid residues, can also be made. The inserted residue(s) can be natural or non-natural, as disclosed above. "Amino acid deletion" refers to the removal of at least one amino acid residue from a predetermined amino acid sequence.

[0049] When referring to a protein, mRNA, or marker as described herein, the terms "level of expression" or "expression level" are generally used interchangeably and generally refer to the detectable amount of a protein, mRNA, or marker in a biological sample. In some embodiments, the detectable amount or detectable level of a protein, mRNA, or marker is associated with the likelihood of a response to an agent as described herein. "Expression" generally refers to the process by which the information contained within a gene is converted into a structure that is present and functional within a cell (e.g., a protein marker such as WSX-1 or IL-27). Thus, as used herein, "expression" can refer to transcription into a polynucleotide, translation into a polypeptide, or modification of a polynucleotide and / or polypeptide (e.g., post-translational modification of a polypeptide). Fragments of a transcribed polynucleotide, a translated polypeptide, or modifications of a polynucleotide and / or polypeptide (e.g., post-translational modification of a polypeptide) are also considered expressed, regardless of whether they are derived from a transcript or degraded transcript generated by alternative splicing, or from post-translational processing of a polypeptide, such as by proteolysis. "Expressed genes" include those that are transcribed into polynucleotides as mRNA and then translated into polypeptides, as well as those that are transcribed into RNA but not translated into polypeptides (e.g., transfer RNA and ribosomal RNA). "Increased expression," "increased expression level," or "increased level" refers to increased expression or increased level of a substance in a sample relative to a control sample, such as an individual or individuals not afflicted with a disease or disorder (e.g., cancer) or an internal control.In some embodiments, increased expression of a substance (e.g., a protein marker such as WSX-1 or IL-27) in a sample refers to an increase in the amount of the substance of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% relative to the amount of the substance in a control sample, as determined by techniques known in the art (e.g., FACS). "Reduced expression," "reduced expression level," or "reduced level" refers to a decrease in expression or level of a substance (e.g., a protein marker) in an individual relative to a control, such as one or more individuals not afflicted with a disease or disorder (e.g., cancer) or an internal control. In some embodiments, reduced expression refers to little or no expression. In some embodiments, a reduction in expression of a substance (e.g., a protein marker) in a sample refers to a decrease in the amount of the substance of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% relative to the amount of the substance in a control sample, as determined by techniques known in the art (e.g., FACS).

[0050] As used herein, the term "angiogenesis" or "angiogenesis" refers to the process of new blood vessels developing from pre-existing blood vessels (Varner et al., (1999) Angiogen. 3:53-60; Mousa et al., (2000) Angiogen. Stim. Inhib. 35:42-44; Kim et al., (2000) Amer. J. Path. 156:1345-1362; Kim et al., (2000) J. Biol. Chem. 275:33920-33928; Kumar et al. (2000) Angiogenesis: From Molecular to Integrative Pharm. 169-180). Endothelial cells from pre-existing blood vessels or circulating endothelial stem cells (Takahashi et al., (1995) Nat. Med. 5:434-438; Isner et al., (1999) J. Clin. Invest. 103:1231-1236) are activated to migrate, proliferate, and differentiate into lumen-bearing structures to form new blood vessels in response to growth factor or hormonal cues or hypoxic or ischemic conditions. During ischemia, such as that occurring in cancer, the need for increased oxygen supply and nutrient delivery induces the secretion of angiogenic factors by the affected tissue. These factors stimulate the formation of new blood vessels. Several additional terms are related to angiogenesis.

[0051] As used herein, the terms "inhibitor," "inhibitor," and "antagonist" can be used interchangeably to refer to any molecule that partially or completely blocks, reduces, inhibits, or neutralizes the biological activity of a native polypeptide disclosed herein. Suitable inhibitors specifically include antagonist antibodies or antibody fragments, fragments or amino acid sequence variants of native polypeptides, peptides, or proteins. In some embodiments, inhibition in the presence of an inhibitor is observed in a dose-dependent manner. In some embodiments, the measured signal (e.g., biological activity) is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% lower than the signal measured using a negative control under comparable conditions. Also disclosed herein are methods for identifying inhibitors suitable for use in the methods of the present disclosure. For example, these methods include, but are not limited to, binding assays such as enzyme-linked immunosorbent assays (ELISAs), ForteBio® systems, radioimmunoassays (RIAs), Meso Scale Discovery assays [e.g., Meso Scale Discovery Electrochemiluminescence (MSD-ECL)], and bead-based Luminex® assays. These assays determine the ability of an inhibitor to bind to a polypeptide of interest (e.g., a receptor or ligand), thereby indicating the inhibitor's ability to inhibit, neutralize, or block the activity of the polypeptide. The effectiveness of an inhibitor can also be determined using a functional assay, such as the ability of an agent to inhibit the function of a polypeptide or agonist. For example, a functional assay can involve contacting a polypeptide with a candidate inhibitor molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide. The potency of an inhibitor is typically measured using an IC50 It is defined by the IC value (the concentration required to inhibit 50% of the agonist response). 50 The lower the value, the more potent the inhibitor and the lower the concentration required to inhibit the maximal biological response.

[0052] As used herein, the phrase "antibody, or antigen-binding portion thereof, that antagonizes human IL-27" refers to an antibody that antagonizes at least one art-recognized activity of human IL-27 (e.g., IL-27 biological activity and / or downstream pathway(s) mediated by IL-27 signaling or other IL-27-mediated function), e.g., a decrease (or reduction) of human IL-27 activity that is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. Additional examples of IL-27 biological activity and / or downstream pathways mediated by IL-27 signaling or other IL-27-mediated function are described in more detail below and elsewhere herein.

[0053] As used herein, the term "anti-IL-27 antagonist antibody" (interchangeably referred to as "anti-IL-27 antibody") refers to an antibody that specifically binds to IL-27 and inhibits the biological activity of IL-27 and / or downstream pathways mediated by IL-27 signaling or other IL-27-mediated functions. Anti-IL-27 antagonist antibodies encompass antibodies that block, antagonize, suppress, inhibit, or reduce IL-27 biological activity (e.g., ligand binding, enzymatic activity), including receptor binding and / or downstream pathways mediated by IL-27 signaling or function, such as eliciting a cellular response to IL-27 or its metabolites. In some embodiments, the anti-IL-27 antagonist antibodies provided by the present disclosure bind to human IL-27 and prevent, block, or inhibit binding of human IL-27 to its cognate or normal receptor (e.g., IL-27 receptor), or one or more receptor subunits [e.g., gp130 and / or IL-27Rα (also known as WSX1 / TCCR)]. In some embodiments, the anti-IL-27 antagonist antibody prevents, blocks, or inhibits the binding of human IL-27 to gp130. In some embodiments, the anti-IL-27 antagonist antibody prevents, blocks, or inhibits the binding of human IL-27 to IL-27Rα. In some embodiments, the anti-IL-27 antagonist antibody prevents, blocks, or inhibits the dimerization of IL-27 monomers. In some embodiments, the anti-IL-27 antibody does not specifically bind to EBI3 monomers. In some embodiments, the anti-IL-27 antibody specifically binds to IL-27p28 monomers. In some embodiments, the anti-IL-27 antibody specifically binds to a discontinuous epitope comprising p28 but does not bind to EBI3 monomers. In some embodiments, the anti-IL-27 antibody inhibits or reduces the phosphorylation of STAT1 and / or STAT3 in cells. In some embodiments, the anti-IL-27 antibody inhibits or reduces inhibition of CD161 expression in the cell (e.g., ameliorate or alleviate IL-27-mediated inhibition of CD161 expression in the cell). In some embodiments, the anti-IL-27 antibody inhibits or reduces PD-L1 expression in the cell.In some embodiments, the anti-IL-27 antibody induces or enhances PD-1-mediated secretion of one or more cytokines from cells. In some embodiments, the anti-IL-27 antibody alters the expression of TIM-3 in cells. In some embodiments, the anti-IL-27 antagonist antibody binds to human IL-27 and stimulates or enhances an anti-tumor response. In some embodiments, the anti-IL-27 antagonist antibody binds to human IL-27 with an affinity of 15 nM or less. In some embodiments, the anti-IL-27 antagonist antibody binds to human IL-27 and comprises a wild-type or mutated IgG1 heavy chain constant region, or a wild-type or mutated IgG4 heavy chain constant region. Examples of anti-IL-27 antagonist antibodies are provided herein.

[0054] As used herein, the term "antibody" refers to a whole antibody comprising two light and two heavy polypeptide chains. Whole antibodies include various antibody isotypes, including IgM, IgG, IgA, IgD, and IgE antibodies. The term "antibody" includes polyclonal, monoclonal, chimeric, humanized, primatized, deimmunized, and fully human antibodies. Antibodies can be produced or derived from any of a variety of mammalian species, including humans, non-human primates (e.g., orangutans, baboons, and chimpanzees), horses, cows, pigs, sheep, goats, dogs, cats, rabbits, guinea pigs, gerbils, hamsters, rats, and mice. Antibodies may be purified or recombinant. As used herein, the terms "antibody fragment," "antigen-binding fragment," or equivalent terms refer to fragments of antibodies that retain the ability to bind to and inhibit the activity of a target antigen (e.g., IL-27). Such fragments include, for example, single-chain antibodies, single-chain Fv fragments (scFv), Fd fragments, Fab fragments, Fab' fragments, or F(ab')2 fragments. An scFv fragment is a single polypeptide chain containing both the heavy and light chain variable regions of the antibody from which the scFv is derived. Additionally, intrabodies, minibodies, triabodies, and diabodies are also included within the definition of antibody and are suitable for use in the methods described herein. See, e.g., Todorovska et al., (2001) J. Immunol. Methods 248(1):47-66; Hudson and Kortt, (1999) J. Immunol. Methods 231(1):177-189; Poljak, (1994) Structure 2(12):1121-1123; Rondon and Marasco, (1997) Annu. Rev. Microbiol. 51:257-283. the disclosures of each of which are incorporated herein by reference.

[0055] As used herein, the term "antibody fragment" also includes single-domain antibodies, such as camelized single-domain antibodies. See, for example, Muyldermans et al., (2001) Trends Biochem. Sci. 26:230-235; Nuttall et al., (2000) Curr. Pharm. Biotech. 1:253-263; Reichmann et al., (1999) J. Immunol. Meth. 231:25-38; PCT Application Publication Nos. WO94 / 04678 and WO94 / 25591, and U.S. Patent No. 6,005,079, all of which are incorporated herein by reference in their entireties. In some embodiments, the present disclosure provides single-domain antibodies comprising two VH domains modified to form single-domain antibodies.

[0056] In some embodiments, the antigen-binding fragment comprises the variable region of a heavy chain polypeptide and the variable region of a light chain polypeptide. In some embodiments, the antigen-binding fragment described herein comprises the CDRs of the light chain polypeptide and the heavy chain polypeptide of the antibody.

[0057] The term "antigen-presenting cell" or "APC" refers to a cell that presents a foreign antigen complexed with MHC on its surface. T cells recognize this complex using the T cell receptor (TCR). Examples of APCs include, but are not limited to, B cells, dendritic cells (DCs), peripheral blood mononuclear cells (PBMCs), monocytes (e.g., THP-1), B lymphoblastoid cells (e.g., C1R.A2, 1518 B-LCL), and monocyte-derived dendritic cells (DCs). Some APCs internalize antigens by phagocytosis or receptor-mediated endocytosis.

[0058] The term "antigen presentation" refers to the process by which APCs capture antigens and make them available for recognition by T cells, for example as components of MHC-I and / or MHC-II complexes.

[0059] As used herein, the term "apoptosis" refers to the process of programmed cell death that occurs in multicellular organisms (e.g., humans). The highly regulated biochemical and molecular events that trigger apoptosis can result in cells undergoing observable and characteristic morphological changes, including membrane blebbing, cell volume reduction, chromosomal DNA condensation and fragmentation, and mRNA decay. A common method for identifying cells, including T cells, undergoing apoptosis is to expose them to a fluorophore-binding protein (annexin V). Annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine in the outer leaflet of the cell membrane, which is an early indicator that cells are undergoing apoptosis.

[0060] As used herein, the term "B cell" (or "B lymphocyte") refers to a type of white blood cell of the lymphocyte subtype. B cells function as the humoral immune component of the adaptive immune system by secreting antibodies. B cells also present antigens and secrete cytokines. B cells differ from two other types of lymphocytes, T cells and natural killer cells, in that they express a B cell receptor (BCR) on their cell membrane. The BCR enables B cells to bind to specific antigens and mount an antibody response against them.

[0061] As used herein, the term "binds to immobilized IL-27" refers to the ability of an antibody of the present disclosure to bind to IL-27 that is, for example, expressed on the surface of a cell or attached to a solid support.

[0062] As used herein, the term "bispecific" or "bifunctional antibody" refers to an artificial hybrid antibody having two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by various methods, including fusion of hybridomas or linking of Fab' fragments. See, for example, Songsivilai & Lachmann, (1990) Clin. Exp. Immunol. 79:315-321; Kostelny et al., (1992) J. Immunol. 148:1547-1553.

[0063] Traditionally, recombinant production of bispecific antibodies has been based on the coexpression of two immunoglobulin heavy / light chain pairs, with the two heavy / light chain pairs having different specificities (Milstein and Cuello, (1983) Nature 305:537-539). Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The heavy chain variable region fusion is preferably with an immunoglobulin heavy chain constant domain, including at least part of the hinge, CH2, and CH3 regions. For further details of exemplary methods currently known for producing bispecific antibodies, see, e.g., Suresh et al., (1986) Methods Enzymol. 121:210; PCT Application WO 96 / 27011; Brennan et al., (1985) Science 229:81; Shalaby et al., J. Exp. Med. (1992) 175:217-225; Kostelny et al., (1992) J. Immunol. 148(5):1547-1553; Hollinger et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Gruber et al., (1994) J. Immunol. 152:5368; and Tutt et al., (1991) J. Immunol. 147:60. Bispecific antibodies also include cross-linked or heteroconjugate antibodies. Heteroconjugate antibodies can be made using any convenient cross-linking method. Suitable cross-linking agents are well known in the art and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques.

[0064] Various techniques for producing and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been generated using leucine zippers. See, e.g., Kostelny et al. (1992) J Immunol 148(5):1547-1553. The leucine zipper peptides from Fos and Jun proteins can be linked to the Fab' portions of two different antibodies by gene fusion. Antibody homodimers can be reduced at the hinge region to form monomers and then reoxidized to form antibody heterodimers. This method can also be used to produce antibody homodimers. The "bispecific antibody" technology described by Hollinger et al. (1993) Proc Natl Acad Sci USA 90:6444-6448 provides an alternative mechanism for generating bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) by a linker that is too short to allow pairing between the two domains on the same chain. Thus, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for generating bispecific antibody fragments using single-chain Fv (scFv) dimers has also been reported. See, e.g., Gruber et al. (1994) J Immunol 152:5368. Alternatively, the antibody may be a "linear antibody," as described, for example, in Zapata et al. (1995) Protein Eng. 8(10):1057-1062. Briefly, these antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1) that form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0065] Antibodies with more than two valencies (eg, trispecific antibodies) are contemplated and are described, for example, in Tutt et al. (1991) J Immunol 147:60.

[0066] The present disclosure also encompasses variant forms of multispecific antibodies, such as the dual variable domain immunoglobulin (DVD-Ig) molecules described in Wu et al. (2007) Nat Biotechnol 25(11): 1290-1297. DVD-Ig molecules are designed such that two different light chain variable domains (VL) from two different parent antibodies are linked in tandem by recombinant DNA technology, either directly or via a short linker, followed by a light chain constant domain. Similarly, the heavy chain contains two different heavy chain variable domains (VH) linked in tandem, followed by a constant domain CH1 and an Fc region. Methods for generating DVD-Ig molecules from two parent antibodies are further described, for example, in PCT Application Publication Nos. WO08 / 024188 and WO07 / 024715. In some embodiments, bispecific antibodies are Fabs-in-Tandem immunoglobulins in which a light chain variable region with a second specificity is fused to a heavy chain variable region of a whole antibody. Such antibodies are described, for example, in International Patent Application Publication No. WO2015 / 103072.

[0067] As used herein, "cancer antigen" or "tumor antigen" refers to (i) tumor-specific antigens, (ii) tumor-associated antigens, (iii) cells expressing tumor-specific antigens, (iv) cells expressing tumor-associated antigens, (v) embryonic antigens on tumors, (vi) autologous tumor cells, (vii) tumor-specific membrane antigens, (viii) tumor-associated membrane antigens, (ix) growth factor receptors, (x) growth factor ligands, and (xi) any other type of antigen or antigen-presenting cell or substance associated with cancer.

[0068] As used herein, the term "cancer-specific immune response" refers to an immune response induced by the presence of tumors, cancer cells, or cancer antigens. In certain embodiments, this response includes the proliferation of cancer antigen-specific lymphocytes. In certain embodiments, this response includes the expression and upregulation of antibodies and T cell receptors, and the formation and release of lymphokines, chemokines, and cytokines. Both the innate immune system and the adaptive immune system interact to initiate antigen responses against tumors, cancer cells, or cancer antigens. In certain embodiments, the cancer-specific immune response is a T cell response.

[0069] The term "cancer" is art-recognized and refers to malignant tumors of epithelial or endocrine tissues, including respiratory, digestive, genitourinary, testicular, breast, prostate, endocrine, and melanoma. The anti-IL-27 antibodies described herein can be used to treat patients with any type of cancer, such as kidney cancer or melanoma, or patients suspected of having or at risk for developing any viral disease. Exemplary cancers include those formed from cervical, lung, prostate, breast, head and neck, colon, and ovarian cancer tissues. The term also includes carcinosarcomas, which include malignant tumors composed of carcinomatous and sarcomatous tissues. "Adenocarcinoma" refers to cancers derived from glandular tissue or in which tumor cells form recognizable glandular structures.

[0070] As used herein, the term "CD112R" refers to a member of the poliovirus receptor-like protein family and is a co-inhibitory receptor for human T cells. CD112R is an inhibitory receptor primarily expressed by T cells and NK cells, competing with the activating receptor CD226 for CD112 binding. The interaction between CD112 and CD112R has higher affinity than the interaction with CD226, thereby effectively controlling CD226-mediated cell activation. Anti-CD112R antagonists that inhibit the interaction with CD112 inhibit direct inhibitory signaling downstream of CD112R and simultaneously promote immune cell activation by increasing the interaction between CD226 and CD112. As used herein, the term "CD112R inhibitor" refers to an agent that disrupts, blocks, or inhibits the biological function or activity of CD112R.

[0071] As used herein, the term "CD137" (or "4-1BB") refers to a member of the tumor necrosis factor (TNF) receptor superfamily. 4-1BB is a costimulatory immune checkpoint molecule primarily for activated T cells. Cross-linking of CD137 enhances T cell proliferation, IL-2 secretion, survival, and cytolytic activity. As used herein, the term "4-1BB agonist" refers to an agent that stimulates, induces, or increases one or more functions of 4-1BB. An exemplary 4-1BB agonist is utomilumab (PF-05082566), a fully human IgG2 monoclonal antibody that targets 4-1BB and stimulates T cells.

[0072] As used herein, "CD161" (alternatively known as killer cell lectin-like receptor subfamily B, member 1 (KLRB1); NK1.1, or NKR-P1A) refers to a member of the C-type lectin superfamily. CD161 is a marker for T cells, and CD161 expression is associated with T cell infiltration into the tumor microenvironment in many different cancer types. CD161 is further described in Fergusson et al., (2014) Cell Reports 9(3):1075-1088, which is incorporated herein by reference in its entirety.

[0073] As used herein, the term "IL-27" or "interleukin-27" refers to the IL-27 cytokine. IL-27 is related to the IL-6 / IL-12 cytokine family and is a heterodimeric cytokine containing a first subunit known as Epstein-Barr virus-induced gene 3 (EBI3; also known as IL-27 subunit β and IL-27B) and a second subunit known as IL-27p28 (also known as IL30, IL-27 subunit α and IL-27A). IL-27 is primarily synthesized by activated antigen-presenting cells, including monocytes, endothelial cells, and dendritic cells (Jankowski et al. (2010) Arch Immunol. Ther. Exp. 58:417-425, Diakowski et al. (2013) Adv. Clin. Exp. Med. (2013) 22(5): 683-691). Although IL-27 can have proinflammatory effects, numerous studies suggest an important role for IL-27 as an immunosuppressant (Shimizu et al. (2006) J. Immunol. 176:7317-7324, Hisada et al. (2004) Cancer Res. 64:1152-1156, Diakowski (2013) supra). IL-27 was initially described as a factor promoting the initiation of Th1 responses, but was later found to fulfill a primary T cell suppressive function by limiting Th1 responses, inhibiting the differentiation of Th2 and Th17 cells, and controlling the development of Tr1 and other T regulatory cell populations (Dietrich et al. (2014) J. Immunol. 192:5382-5389). In addition to its role as an immunomodulator, IL-27 also regulates angiogenesis, hematopoiesis, and osteoclastogenesis.

[0074] IL-27 signals through a heterodimeric type I cytokine receptor (IL-27 receptor or IL-27R) that contains a first subunit called WSX-1 (also known as IL-27 receptor subunit α, IL-27RA, T-cell cytokine receptor type 1 (TCCR), and cytokine receptor-like 1 (CRL1)) and a second subunit called gp130 (also known as interleukin-6 signaling factor (IL6ST), interleukin-6 receptor subunit β (IL-6RB), and oncostatin M receptor). gp130 is also a receptor subunit for IL-6 family cytokines (Liu et al. (2008) Scan. J. Immunol. 68:22-299, Diakowski (2013) supra). IL-27 signaling through IL-27R activates multiple signaling cascades, including the JAK-STAT pathway and the p38MAPK pathway.

[0075] EBI3 is also thought to have biological functions independent of p28 or IL-27 heterodimers. For example, EBI3 also interacts with p35 to form the heterodimeric cytokine IL-35 (Yoshida et al. (2015) Annu. Rev Immunol. 33:417-43), and has been shown to be selectively overexpressed in certain cell types without a corresponding increase in p28 or IL-27 (Larousserie et al. (2005) Am. J. Pathol. 166(4):1217-28).

[0076] The amino acid sequence of an exemplary human EBI3 protein is provided in SEQ ID NO: 1 (NCBI Reference Sequence: NP_005746.2; N-mtpqlllalvlwascppcsgrkgppaaltlprvqcrasrypiavdcswtlppapnstspvsfiatyrlgmaarghswpclqqtptstsctitdvqlfsmapyvlnvtavhpwgssssfvpfitehiikpdppegvrlsplaerqlqvqweppgswpfpeifslkywirykrqgaarfhrvgpieatsfilravrpraryyvqvaaqdltdygelsdwslpatatmslgk-C). The amino acid sequence of an exemplary human p28 protein is provided in SEQ ID NO: 2 (NCBI Reference Sequence: NP_663634.2; N-mgqtagdlgwrlsllllplllvqagvwgfprppgrpqlslqelrreftvslhlarkllsevrgqahrfaeshlpgvnlyllplgeqlpdvsltfqawrrlsdperlcfisttlqpfhallgglgtqgrwtnmermqlwamrldlrdlqrhlrfqvlaagfnlpeeeeeeeeeeeeeeerkgllpgalgsalqgpaqvswpqllstyrllhslelvlsravrellllskaghsvwplgfptlspqp-C).The amino acid sequence of an exemplary human WSX1 protein is provided in SEQ ID NO: 3 (NCBI Reference Sequence: NP_004834.1; N--C).The amino acid sequence of an exemplary human gp130 protein is provided in SEQ ID NO: 4 (NCBI Reference Sequence: NP_002175.2; N--C).

[0077] As used herein, the term "combined positive score" or "CPS" refers to the number of positive tumor cells, lymphocytes, and macrophages divided by the total number of viable tumor cells multiplied by 100.

[0078] As used herein, the term "competition," when used in the context of antigen binding proteins (e.g., immunoglobulins, antibodies, or antigen-binding fragments thereof) that compete for binding to the same epitope, refers to an interaction between antigen binding proteins as determined by an assay (e.g., competitive binding assay; cross-blocking assay) in which a test antigen binding protein (e.g., test antibody) inhibits (e.g., reduces or blocks) the specific binding of a reference antigen binding protein (e.g., reference antibody) to a common antigen (e.g., IL-27 or a fragment thereof).

[0079] A polypeptide or amino acid sequence "derived from" a specified polypeptide or protein refers to the origin of the polypeptide. Preferably, a polypeptide or amino acid sequence derived from a particular sequence has an amino acid sequence essentially identical to that sequence, or a portion thereof, where the portion consists of or originates from at least 10-20 amino acids, preferably at least 20-30 amino acids, and more preferably at least 30-50 amino acids, discernible by one of skill in the art. A polypeptide derived from another peptide may have one or more mutations relative to the starting polypeptide, such as a mutation having one or more amino acid residues substituted with another amino acid residue, or an insertion or deletion of one or more amino acid residues.

[0080] Polypeptides can include amino acid sequences that do not occur in nature. Such variants necessarily have less than 100% sequence identity or similarity with the starting molecule. In certain embodiments, variants have about 75% to less than 100% amino acid sequence identity or similarity, more preferably about 80% to less than 100%, even more preferably about 85% to less than 100%, even more preferably about 90% to less than 100% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%), and most preferably about 95% to less than 100% amino acid sequence identity or similarity with the amino acid sequence of the starting polypeptide, e.g., over the length of the variant molecule.

[0081] In certain embodiments, the antibodies of the present disclosure are encoded by nucleotide sequences. The nucleotide sequences of the present disclosure may be useful for a number of applications, including cloning, gene therapy, protein expression and purification, mutagenesis, DNA vaccination of a host in need thereof, antibody production for, for example, passive immunization, PCR, primer and probe production, etc.

[0082] Those skilled in the art will also understand that antibodies suitable for use in the methods disclosed herein may be modified to differ in sequence from the native or natural sequence from which they are derived while retaining the desired activity of the native sequence. For example, nucleotide or amino acid substitutions leading to conservative substitutions or changes in "non-essential" amino acid residues may be made. Mutations may be introduced by standard techniques such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0083] Antibodies suitable for use in the methods disclosed herein may contain conservative amino acid substitutions at one or more amino acid residues, for example, at essential or non-essential amino acid residues. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues with similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, non-essential amino acid residues in a binding polypeptide are preferably replaced with another amino acid residue from the same side chain family. In certain embodiments, a string of amino acids can be replaced with a structurally similar string that differs in the order and / or composition of the side chain family members. Alternatively, in certain embodiments, mutations can be randomly introduced along all or part of the coding sequence, such as by saturation mutagenesis, and the resulting mutations can be incorporated into a binding polypeptide of the present disclosure and screened for their ability to bind to a desired target.

[0084] As used herein, the term "antigen cross-presentation" refers to the presentation of exogenous protein antigens to T cells via MHC class I and class II molecules on APCs.

[0085] As used herein, the term "cross-reactivity" refers to the ability of an antibody of the present disclosure to bind to IL-27 of a different species. For example, an antibody of the present disclosure that binds to human IL-27 may also bind to IL-27 of another species. As used herein, cross-reactivity is measured by detecting specific reactivity with purified antigen in a binding assay (e.g., SPR, ELISA), or by binding to or functionally interacting with cells that physiologically express IL-27. Methods for determining cross-reactivity include standard binding assays as described herein, such as Biacore™ surface plasmon resonance (SPR) analysis using a Biacore™ 2000 SPR instrument (Biacore AB, Uppsala, Sweden), or flow cytometry techniques.

[0086] As used herein, the term "cytotoxic T lymphocyte (CTL) response" refers to an immune response induced by cytotoxic T cells. CTL responses are primarily CD8 + Mediated by T cells.

[0087] As used herein, the term "dendritic cell" or "DC" refers to a type of antigen-presenting cell, a bone marrow (BM)-derived white blood cell that is the most potent type of antigen-presenting cell. DCs capture and process antigens, converting proteins into peptides that are displayed on major histocompatibility complex (MHC) molecules for recognition by T cells. DCs are heterogeneous, e.g., myeloid DCs and plasmacytoid DCs. While all DCs are capable of uptake, processing, and presentation of antigens to naive T cells, DC subtypes possess distinct markers and differ in their location, migration route, detailed immune function, and dependence on infectious or inflammatory stimuli for their generation. During the development of the adaptive immune response, DC phenotype and function play a role in the initiation of tolerance, memory, and polarization into T helper 1 (Th1), Th2, and Th17.

[0088] As used herein, the term "dendritic cell activation" refers to the transition from immature dendritic cells to mature dendritic cells. Activated dendritic cells include mature dendritic cells and dendritic cells in the transitional process. Expression of CD80 and CD86, which induce costimulatory signals, increases upon activation. Mature human dendritic cells are cells that are positive for CD40, CD80, CD86, and HLA class II (e.g., HLA-DR). Immature dendritic cells can be distinguished from mature dendritic cells based on markers selected from the group consisting of CD80 and CD86. Immature dendritic cells are weakly positive, or preferably negative, for these markers, while mature dendritic cells are positive. The identification of mature dendritic cells is routinely performed by those skilled in the art, and methods for measuring the above-mentioned markers and their expression are also well known to those skilled in the art.

[0089] As used herein, "EC 50 The term "antibody" refers to the concentration of an antibody or antigen-binding portion thereof that induces 50% of the maximal response, i.e., a response halfway between the maximal response and baseline, in either an in vitro or in vivo assay.

[0090] As used herein, the term "effective amount" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The term "therapeutically effective amount" is defined as an amount sufficient to cure or at least partially arrest the disease and its complications in patients already suffering from the disease. The amount effective for this use will vary depending on the severity of the disease being treated and the general condition of the patient's own immune system.

[0091] As used herein, the term "epitope" or "antigenic determinant" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds. The term "epitope mapping" refers to the process or method of identifying the binding site, i.e., epitope, of an antibody or its antigen-binding fragment on its target protein antigen. Epitope mapping methods and techniques are provided herein. Epitopes can be formed from contiguous amino acids or non-contiguous amino acids juxtaposed by tertiary folding of a protein. Epitopes formed from contiguous amino acids are typically retained when exposed to denaturing solvents, while epitopes formed by tertiary folding are typically lost when treated with denaturing solvents. An epitope typically contains at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. Methods for determining which epitope is bound by a given antibody (i.e., epitope mapping) are well known in the art and include, for example, immunoblotting and immunoprecipitation assays, in which overlapping or consecutive peptides from IL-27 are tested for reactivity with a given anti-IL-27 antibody. Methods for determining the spatial conformation of epitopes include techniques in the art and described herein, such as X-ray crystallography and two-dimensional nuclear magnetic resonance (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, G. E. Morris, Ed. (1996)).

[0092] Also encompassed by the present disclosure are antibodies that bind to epitopes on IL-27 that include all or part of (e.g., the same or overlapping regions, or regions between or spanning) the epitopes recognized by the specific antibodies described herein.

[0093] Also encompassed by the present disclosure are antibodies that bind to the same epitope and / or antibodies that compete with the antibodies described herein for binding to human IL-27. Antibodies that recognize the same epitope or that compete for binding can be identified using routine techniques. Such techniques include, for example, immunoassays that demonstrate the ability of one antibody to inhibit the binding of another antibody to a target antigen, i.e., competitive binding assays. Competitive binding is determined in an assay in which the immunoglobulin under test inhibits the specific binding of a reference antibody to a common antigen, such as IL-27. Many types of competitive binding assays are known, including, for example, solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid-phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid-phase direct label assays, solid-phase direct label sandwich assays (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid-phase direct label RIA using I-125 labels (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid-phase direct biotin-avidin EIA (see Cheung et al., Virology 176:546 (1986)); (1990)); and direct labeling RIA (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)). Typically, such assays involve the use of purified antigen bound to a solid surface or cells bearing either an unlabeled test immunoglobulin and a labeled reference immunoglobulin. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cells in the presence of the test immunoglobulin. Typically, the test immunoglobulin is present in excess.Typically, when a competing antibody is present in excess, it inhibits specific binding of a reference antibody to a common antigen by at least 50-55%, 55-60%, 60-65%, 65-70%, 70-75% or more.

[0094] Other techniques include epitope mapping, such as X-ray analysis of crystals of antigen-antibody complexes, which provides atomic resolution of epitopes, and mass spectrometry combined with hydrogen / deuterium (H / D) exchange to study the conformation and dynamics of antigen-antibody interactions. Other methods monitor antibody binding to antigen fragments or mutant variations of antigens, where loss of binding due to modification of amino acid residues within the antigen sequence is often considered indicative of epitope components. Additionally, computational combinatorial methods for epitope mapping can also be used. These methods rely on the ability of a subject antibody to affinity isolate specific short peptides from a combinatorial phage-display peptide library. These peptides are then considered leads to define the epitope corresponding to the antibody used to screen the peptide library. For epitope mapping, computational algorithms have also been developed that have been shown to map conformationally discontinuous epitopes.

[0095] As used herein, the term "Fc-mediated effector function" or "Fc effector function" refers to the primary function of an antibody and other biological activities of the antibody besides its intended target. For example, the effector function of a therapeutic agnostic antibody is a biological activity other than the activation of a target protein or pathway. Examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors); lack of activation of platelets expressing Fc receptors; and activation of B cells. Many effector functions are initiated by Fc binding to Fcγ receptors. In some embodiments, tumor antigen-targeting antibodies have effector functions, such as ADCC activity. In some embodiments, the tumor antigen-targeting antibodies described herein comprise a variant constant region having an increased effector function (e.g., an increased ability to mediate ADCC) compared to an unmodified form of the constant region.

[0096] As used herein, the term "Fc receptor" refers to a polypeptide found on the surface of immune effector cells and coupled to the Fc region of an antibody. In some embodiments, the Fc receptor is an Fcγ receptor. There are three subclasses of Fcγ receptors: FcγRI (CD64), FcγRII (CD32), and FcγcRIII (CD16). All four IgG isotypes (IgG1, IgG2, IgG3, and IgG4) bind to and activate Fc receptors FcγRI, FcγRIIA, and FcγRIIIA. FcγRIIB is an inhibitory receptor; therefore, antibodies bound to this receptor do not activate complement and cellular responses. FcγRI is a high-affinity receptor that binds to IgG in monomeric form, while FcγRIIA and FcγRIIA are low-affinity receptors that only bind to IgG in multimeric form and have weakly low affinity. Antibody binding to Fc receptors and / or C1q is determined by specific residues or domains within the Fc region. Binding also depends on residues located within the hinge region and CH2 portion of the antibody. In some embodiments, the agonistic and / or therapeutic activity of the antibodies described herein depends on the binding of the Fc region to an Fc receptor (e.g., FcγR). In some embodiments, the agonistic and / or therapeutic activity of the antibodies described herein is enhanced by the binding of the Fc region to an Fc receptor (e.g., FcγR).

[0097] A list of certain Fc receptor sequences used in this disclosure is provided below in Table 1B.

[0098] As used herein, the term "GBP5" or "guanylate-binding protein 5" refers to a member of guanylate-binding proteins (GBPs). GBP5 has various biological functions, and the encoded protein functions as an activator of NLRP3 inflammasome assembly and plays a role in innate immunity and inflammation. (See, for example, Li, Xiang et al. Frontiers in Genetics vol. 13 984615. 30 Sep. 2022, which is incorporated herein by reference in its entirety.)

[0099] As used herein, the term "glycosylation pattern" is defined as the pattern of carbohydrate units covalently attached to a protein, more particularly to an immunoglobulin protein. If a person skilled in the art recognizes that the glycosylation pattern of a heterologous antibody is more similar to the glycosylation pattern in the species of the non-human transgenic animal than to the species from which the transgenic CH gene is derived, the glycosylation pattern of the heterologous antibody can be characterized as being substantially similar to the glycosylation pattern naturally occurring on antibodies produced by the species of the non-human transgenic animal.

[0100] As used herein, the term "human antibody" includes antibodies having variable and constant regions (if present) of human germline immunoglobulin sequences. The human antibodies of the present disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo) (see, e.g., Lonberg et al., (1994) Nature 368(6474): 856-859; Lonberg, (1994) Handbook of Experimental Pharmacology 113:49-101; Lonberg & Huszar, (1995) Intern. Rev. Immunol. 13:65-93, and Harding & Lonberg, (1995) Ann. NY Acad. Sci. 764:536-546). However, the term "human antibody" does not include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences (i.e., humanized antibodies).

[0101] As used herein, the term "heterologous antibody" is defined in relation to the transgenic non-human organism producing such an antibody. This term refers to an antibody having an amino acid sequence or encoding nucleic acid sequence that corresponds to that found in an organism not consisting of a transgenic non-human animal, generally an organism derived from a species other than the transgenic non-human animal.

[0102] As used herein, the term "interferon regulatory factor 1" or "IRF1" refers to a nuclear factor that binds to and activates the promoter of type I interferon genes. IRF1 is an early target gene downstream of IFNγ signaling and regulates IFNγ-mediated gene induction. IRF-1 is a member of the interferon regulatory factor (IRF) family. IRF-1 was first identified in 1988 as a transcription factor capable of inducing the expression of the gene interferon beta (IFN-B). See Dou, Lei et al. Human Immunology Vol. 75,11 (2014): 1110-4, which is incorporated herein by reference in its entirety.

[0103] As used herein, the term "immune cell" refers to any cell of the human immune system. The term "immune cell" includes, but is not limited to, lymphocytes (e.g., T cells, B cells, and tumor-infiltrating lymphocytes (TILs)), macrophages, basophils, eosinophils, neutrophils, monocytes, and natural killer (NK) cells.

[0104] The terms "inducing an immune response" and "enhancing an immune response" are used interchangeably and refer to the stimulation of an immune response (i.e., passive or adaptive) to a particular antigen. The term "induction" when used in reference to inducing CDC or ADCC refers to the stimulation of a specific direct cell-killing mechanism.

[0105] As used herein, the term "immunogenic cell death" (also known as "immunogenic apoptosis") refers to a mode of cell death associated with the activation of one or more signaling pathways that induce the pre-death expression and release of damage-associated molecular pattern (DAMP) molecules (e.g., adenosine triphosphate, ATP) from tumor cells, resulting in increased immunogenicity of the tumor cells and their death in an immunogenic manner (e.g., by phagocytosis). As used herein, the term "immunogenic cell death inducer" refers to a chemical, biological, or pharmacological agent that induces the process, pathway, or aspect of immunogenic cell death.

[0106] As used herein, the terms "inhibit," "reduce," or "block" (e.g., referring to the inhibition or reduction of human IL-27-mediated phosphorylation of STAT1 and / or STAT3 in a cell) are used interchangeably and encompass both partial and complete inhibition / blocking. Inhibition / blocking of IL-27 reduces or alters the normal level or type of activity that occurs without inhibition or blockage. Inhibition and blocking are also intended to include a measurable reduction in the binding affinity of IL-27 when contacted with an anti-IL-27 antibody compared to IL-27 not contacted with the anti-IL-27 antibody, e.g., inhibiting IL-27 binding by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0107] As used herein, the terms "inhibiting angiogenesis," "reducing angiogenesis," and "reducing angiogenesis" refer to reducing the level of angiogenesis in a tissue to at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or less than the amount in a corresponding control tissue, and most preferably to a level similar to that observed in the control tissue.

[0108] As used herein, "inhibiting proliferation" (e.g., with respect to cells) is intended to include any measurable decrease in proliferation of the cells, e.g., an inhibition of proliferation of the cells by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%.

[0109] As used herein, a subject "in need of prevention," "in need of treatment," or "in need of" refers to a subject who, as determined by a competent medical practitioner (e.g., in the case of a human, a physician, nurse, or advanced practice nurse practitioner; in the case of a non-human mammal, etc., a veterinarian), would reasonably benefit from a given treatment (e.g., treatment with a composition comprising an anti-IL-27 antibody).

[0110] The term "in vivo" refers to a process that takes place inside a living organism.

[0111] As used herein, the term "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to human IL-27 is substantially free of antibodies that specifically bind to antigens other than IL-27). However, an isolated antibody that specifically binds to an epitope may exhibit cross-reactivity to other IL-27 proteins from different species. However, the antibody continues to exhibit specific binding to human IL-27 in the specific binding assays described herein. Furthermore, an isolated antibody is typically substantially free of other cellular material and / or chemicals. In some embodiments, a combination of "isolated" antibodies with different IL-27 specificities is combined in a well-defined composition.

[0112] As used herein, the term "isolated nucleic acid molecule" refers to an antibody or antibody portion that binds IL-27 (e.g., V H , V L, CDR3), where the nucleotide sequence encoding the antibody or antibody portion does not include other nucleotide sequences encoding antibodies or antibody portions that bind to antigens other than IL-27, and other sequences may naturally flank the nucleic acid in human genomic DNA. For example, a sequence selected from the sequences set forth in Table 1A may be selected from the heavy chain (V , CDR4) of an anti-IL-27 antibody monoclonal antibody described herein. H ) and light chain (V L ) corresponds to a nucleotide sequence containing the variable region.

[0113] As used herein, "isotype" refers to the antibody class (e.g., IgM or IgG1) encoded by the heavy chain constant region genes. In some embodiments, the human monoclonal antibodies of this disclosure are of the IgG1 isotype. In some embodiments, the human monoclonal antibodies of this disclosure are of the IgG2 isotype. In some embodiments, the human monoclonal antibodies of this disclosure are of the IgG3 isotype. In some embodiments, the human monoclonal antibodies of this disclosure are of the IgG4 isotype. As will be apparent to those of skill in the art, identifying antibody isotypes (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE) is routine in the art and generally involves a combination of known antibodies, published Fc variant sequences, and sequence alignment with conserved sequences.

[0114] As used herein, the term "isotype switching" refers to the phenomenon in which the class or isotype of an antibody changes from one Ig class to one of the other Ig classes.

[0115] As used herein, "KD" or "K D The term "K" refers to the equilibrium dissociation constant of the binding reaction between an antibody and an antigen. DThe value of K is the ratio of the antibody off-rate constant (kd) to the antibody on-rate constant (ka). D The value is inversely proportional to the binding affinity of the antibody to the antigen. D The smaller the value, the higher the affinity of the antibody for the antigen. Affinity is the strength of binding of a single molecule to its ligand and is typically expressed as the equilibrium dissociation constant (K D ) and is used to assess and rank the strength of bimolecular interactions.

[0116] As used herein, "kd" or "k d (or "koff" or "k off The term k is intended to refer to the off-rate constant for dissociation of an antibody from the antibody / antigen complex. d The value of is a numerical representation of the rate at which the complex disintegrates or dissociates per second, and is expressed in units of sec -1 is.

[0117] As used herein, "ka" or "k a (or "kon" or "k on The term ka is intended to refer to the on-rate constant for the association of an antibody with an antigen. The value of ka is a numerical representation of the number of antibody / antigen complexes formed per second in a 1 molar (1M) solution of antibody and antigen, and is expressed in units of M -1 sec -1 is.

[0118] As used herein, the term "leukocyte" refers to a type of white blood cell that is involved in defending the body against infectious organisms and foreign substances. Leukocytes are produced in the bone marrow. There are five main types of white blood cells, divided into two main groups: polymorphonuclear leukocytes (neutrophils, eosinophils, basophils) and mononuclear leukocytes (monocytes and lymphocytes).

[0119] As used herein, the term "lymphocyte" refers to a type of white blood cell or leukocyte that is involved in the body's immune defenses. There are two main types of lymphocytes: B cells and T cells.

[0120] As used herein, the terms "linked," "fused," or "fusion" are used interchangeably. These terms refer to the joining of two or more elements, components, or domains to one another by any means, including chemical or recombinant means. Methods of chemical conjugation (e.g., the use of heterobifunctional cross-linkers) are known in the art.

[0121] As used herein, " local administration " or " local delivery " refers to delivery that does not rely on transporting composition or drug to intended target tissue or site through vascular system.For example, composition can be delivered by injection or implantation of composition or drug, or by injection or implantation of device containing composition or drug.After local administration near target tissue or site, composition or drug, or one or more components thereof, can diffuse to intended target tissue or site.

[0122] As used herein, "MHC molecule" refers to two types of molecules: MHC class I and MHC class II. MHC class I molecules present antigens to specific CD8+ T cells, and MHC class II molecules present antigens to specific CD4+ T cells. Antigens delivered exogenously to APCs are processed primarily for binding to MHC class II. In contrast, antigens delivered endogenously to APCs are processed primarily for binding to MHC class I.

[0123] As used herein, the term "monoclonal antibody" refers to an antibody that exhibits a single binding specificity and affinity for a specific epitope. Thus, the term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity and has a variable region and any constant region derived from a human germline immunoglobulin sequence. In some embodiments, human monoclonal antibodies are produced by hybridomas that contain B cells obtained from transgenic non-human animals, such as transgenic mice, whose genomes contain human heavy chain transgenes and light chain transgenes fused to immortalized cells.

[0124] As used herein, the term "monocyte" refers to a type of white blood cell that can differentiate into macrophages and dendritic cells to trigger an immune response.

[0125] As used herein, the term "natural killer (NK) cells" refers to a type of cytotoxic lymphocyte. These are large, usually granular, non-T, non-B lymphocytes that kill certain tumor cells and play an important role in innate immunity against viruses and other intracellular pathogens, as well as antibody-dependent cell-mediated cytotoxicity (ADCC).

[0126] As used herein, the term "naturally occurring" as applied to an object refers to the fact that the object can be found in nature. For example, a polypeptide or polynucleotide sequence that can be isolated from a natural source and exists in an organism (including a virus) that has not been artificially modified in a laboratory is naturally occurring.

[0127] As used herein, the term "non-switched isotype" refers to the heavy chain isotype class generated when isotype switching has not occurred, and the CH gene encoding the non-switched isotype is typically the first CH gene immediately downstream of the functionally rearranged VDJ gene. Isotype switching is classified as classical isotype switching or non-classical isotype switching. Classical isotype switching occurs through recombination events involving at least one switch sequence region within the transgene. Non-classical isotype switching occurs, for example, in human σ μ and human Σ μ Alternative non-classical switching events, such as intergenic and interchromosomal recombination, among others, can occur, resulting in isotype switching.

[0128] As used herein, the term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides, and their polymers, in either single-stranded or double-stranded form.Unless otherwise limited, this term encompasses nucleic acids that contain known analogs of natural nucleotides, which have similar binding properties to reference nucleic acids and are metabolized in a manner similar to natural nucleotides.Unless otherwise indicated, a specific nucleic acid sequence also implicitly encompasses its conservatively modified variants (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more (or all) selected codons is replaced with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., Biol. Chem. 260:2605-2608, 1985; and Cassol et al., 1992; Rossolini et al., Mol. Cell. Probes 8:91-98, 1994). For arginine and leucine, modifications to the second base can also be conservative. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene.

[0129] As used herein, a polynucleotide can be composed of any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA, or modified RNA or DNA. For example, a polynucleotide can be composed of single-stranded and double-stranded DNA, DNA that is a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, RNA that is a mixture of single-stranded and double-stranded regions, or hybrid molecules containing DNA and RNA that may be single-stranded, more typically double-stranded, or a mixture of single-stranded and double-stranded regions. Furthermore, a polynucleotide can be composed of RNA or DNA, or triple-stranded regions containing both RNA and DNA. A polynucleotide can also contain one or more modified bases, or DNA or RNA backbones modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. Various modifications can be made to DNA and RNA, and therefore "polynucleotide" includes chemically, enzymatically, or metabolically modified forms.

[0130] A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of that coding sequence. With respect to transcriptional regulatory sequences, operably linked means that the linked DNA sequences are contiguous and, where necessary to join two protein-coding regions, contiguous and in reading frame. With respect to switch sequences, operably linked indicates that the sequences are capable of undergoing switch recombination.

[0131] As used herein, the terms "parenteral administration," "parenterally administered," and other grammatically equivalent phrases refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intranasal, intraocular, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, intracerebral, intracranial, intracarotid, and intrasternal injection and infusion.

[0132] As used herein, the term "patient" includes human and other mammalian subjects receiving prophylactic or therapeutic treatment.

[0133] As used herein, the term "PD-1 antagonist" refers to any chemical compound or biological molecule that inhibits the PD-1 signaling pathway or inhibits PD-1 function in a cell (e.g., an immune cell). In some embodiments, the PD-1 antagonist blocks the binding of PD-L1 to PD-1 and / or PD-L2 to PD-1. In some embodiments, the PD-1 antagonist specifically binds to PD-1. In some embodiments, the PD-1 antagonist specifically binds to PD-L1.

[0134] The term "percent identity," in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that have a specified percentage of the same nucleotides or amino acid residues when compared and aligned for maximum correspondence, as determined using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN, or other algorithms available to those skilled in the art) or by visual inspection. Depending on the application, the "percent identity" may be over a region of the sequences being compared, such as a functional domain, or over the entire length of the two sequences being compared. In sequence comparison, typically, one sequence serves as a reference sequence to which a test sequence is compared. When using a sequence comparison algorithm, the test sequence and the reference sequence are input into a computer, subsequence coordinates are designated as necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the designated program parameters.

[0135] Optimal alignment of sequences for comparison can be performed, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computer implementations of these algorithms (GAP, BESTFIT, FASTA, TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).

[0136] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the website of the National Center for Biotechnology Information.

[0137] As generally used herein, "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and / or body fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0138] As used herein, "pharmaceutically acceptable carrier" refers to and includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. The compositions may include pharmaceutically acceptable salts, such as acid addition salts or base addition salts (see, e.g., Berge et al. (1977) J Pharm Sci 66:1-19).

[0139] As used herein, the terms "polypeptide," "peptide," and "protein" are used interchangeably to refer to polymers of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as to naturally occurring and unnatural amino acid polymers.

[0140] As used herein, the term "prevention" when used in reference to a condition refers to the administration of a composition that reduces the frequency or delays the onset of symptoms of the condition in a subject compared to a subject not administered the composition.

[0141] As used herein, the terms "purified" or "isolated" when applied to any protein (antibody or fragment) described herein refers to a polypeptide that has been separated or purified from components that naturally accompany it (e.g., proteins or other naturally occurring biological or organic molecules), such as other proteins, lipids, and nucleic acids, in the prokaryotic organism that expresses the protein. Typically, a polypeptide is purified when it constitutes at least 60% (e.g., at least 65, 70, 75, 80, 85, 90, 92, 95, 97, or 99%) by weight of the total protein in a sample.

[0142] As used herein, the term "programmed cell death protein 1" or "PD-1" refers to the programmed cell death protein 1 polypeptide, an immunosuppressive receptor belonging to the CD28 family and encoded by the PDCD1 gene in humans. Other names or synonyms for PD-1 include PDCD1, PD1, CD279, and SLEB2. PD-1 is primarily expressed in vivo on preactivated T cells, B cells, and myeloid cells and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one shared epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank accession number AAC51773.

[0143] As used herein, the term "programmed death ligand-1" or "PD-L1" refers to one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2), which downregulates T cell activation and cytokine secretion when bound to PD-1. Alternative names and synonyms for PD-L1 include PDCD1L1, PDL1, B7H1, B7-4, CD274, and B7-H. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, as well as analogs that share at least one shared epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank accession number Q9NZQ7.

[0144] PD-1 is known as an immunosuppressive protein that negatively regulates TCR signaling (Ishida, Y. et al. (1992) EMBO J. 11:3887-3895; Blank, C. et al. (Epub 2006 Dec. 29) Immunol. Immunother. 56(5):739-745). The interaction of PD-1 and PD-L1 functions as an immune checkpoint and can lead to a decrease in T cell receptor-mediated proliferation (Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100). Immune suppression can be reversed by inhibiting the local interaction of PD-1 with PD-L1 or PD-L2, and the effect is additive when the interaction of PD-1 with PD-L2 is also inhibited (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0145] In some cancers, tumor survival and growth are maintained by tumor-mediated immune checkpoint regulation. This regulation can disrupt the function of the anti-cancer immune system. For example, recent studies suggest that expression of immune checkpoint receptor ligands, such as PD-L1 or PD-L2, by tumor cells can promote immune evasion by downregulating immune system activity in the tumor microenvironment, particularly by suppressing T cells. PD-L1 is abundantly expressed in various human cancers (Dong et al., (2002) Nat Med 8:787-789). PD-L1 receptor PD-1 is expressed on lymphocytes (e.g., activated T cells) and is normally involved in downregulating the immune system and promoting self-tolerance, particularly by suppressing T cells. However, when PD-1 receptors expressed on T cells bind to their cognate PD-L1 ligands on tumor cells, the resulting suppression of T cells contributes to impaired immune responses against tumors (e.g., reduction of tumor-infiltrating lymphocytes or establishment of immune evasion by cancer cells).

[0146] For example, in large sample sets of ovarian, renal, colorectal, pancreatic, and liver cancers, and melanoma, PD-L1 expression has been shown to correlate with poor prognosis and reduce overall survival regardless of subsequent treatment (e.g., Dong et al., (2002) Nat Med 8(8):793-800; Yang et al., (2008) Invest Ophthalmol Vis Sci 49(6):2518-2525; Ghebeh et al., (2006) Neoplasia 8:190-198; Hamanishi et al., (2007) Proc Nat Acad Sci USA 104:3360-3365; Thompson et al., (2006) Clin Genitourin Cancer 5:206-211; Nomi et al., (2005) Clin Cancer Res 11:2947-2953; Inman et al., (2007) Cancer 109:1499-1505; Shimauchi et al., (2007) Int J Cancer 121:2585-2590; Gao et al., (2009) Clin Cancer Res 15:971-979; Nakanishi et al., (2007) Cancer Immunol Immunother 56:1173-1182; see Hino et al., (2010) Cancer 116(7):1757-1766). Similarly, PD-1 expression on tumor lymphocytes has been found to mark dysfunctional T cells in breast cancer (Kitano et al., (2017) ESMO Open 2(2):e000150) and melanoma (Kleffel et al., (2015) Cell 162(6):1242-1256). PD-1 antagonists, such as those that affect the function of the PD-1 / PD-L1 / PD-L2 signaling axis and / or disrupt the interaction between PD-1 and PD-L1 and / or PD-L2, have been developed and represent a novel class of antitumor inhibitors that function through modulation of immune cell-tumor cell interactions.

[0147] As used herein, the term "recombinant host cell" (or simply "host cell") is intended to refer to a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but to the progeny of such a cell. Because certain modifications may occur in the progeny, either due to mutational or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0148] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated, or isolated by recombinant means, including, for example, (a) antibodies isolated from or hybridomas prepared from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes, (b) antibodies isolated from host cells, e.g., transfectomas, transformed to express the antibody, (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated, or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies are composed of variable and constant regions utilizing specific human germline immunoglobulin sequences encoded by germline genes, but including subsequent rearrangements and mutations that occur, for example, during antibody maturation. As known in the art (see, e.g., Lonberg (2005) Nature Biotech. 23(9):1117-1125), variable regions contain antigen-binding domains, which are encoded by various genes that rearrange to form antibodies specific to foreign antigens. In addition to rearrangement, variable regions can be further modified by multiple single amino acid changes (called somatic mutations or hypermutations) to increase the affinity of the antibody for the foreign antigen. The constant regions change in further response to the antigen (i.e., isotype switching). Thus, rearranged and somatically mutated nucleic acid molecules encoding light and heavy immunoglobulin polypeptides in response to an antigen may not have sequence identity with the original nucleic acid molecule, but instead are substantially identical or similar (i.e., have at least 80% identity).

[0149] As used herein, the term "reference antibody" (used interchangeably with "reference mAb") or "reference antigen-binding protein" refers to an antibody or antigen-binding fragment thereof that binds to a particular epitope on IL-27 and is used to establish a relationship between itself and one or more different antibodies, the relationship being the binding of the reference antibody and the one or more different antibodies to the same epitope on IL-27. As used herein, the term refers to an anti-IL-27 antibody that is useful as a competitor in a test or assay such as those described herein (e.g., a competitive binding assay), which is useful for discovering, identifying, or developing one or more different antibodies that bind to the same epitope.

[0150] As used herein, the terms "specific binding," "selective binding," "selectively binds," and "specifically binds" refer to an antibody that binds to an epitope on a given antigen. Typically, the antibody binds to an epitope of about 10% or more as measured by surface plasmon resonance (SPR) technology on a BIACORE 2000 instrument using recombinant human IL-27 as the analyte and the antibody as the ligand. -6 Less than m, e.g., about 10 -7 , 10 -8 M, 10 - 9M or 10 -10 M or lower equilibrium dissociation constant (K D ) and binds to a predetermined antigen with an affinity at least two-fold higher than its affinity to bind to a nonspecific antigen other than the predetermined antigen or a closely related antigen (e.g., BSA, casein). In certain embodiments, an antibody that specifically binds to IL-27 has an affinity of about 100 nM (10) as measured by surface plasmon resonance (SPR) technology on a BIACORE 2000 instrument using recombinant human IL-27 as the analyte and the antibody as the ligand. -7 Less than 50 nM (5 x 10 -8 M), approximately 15 nM (1.5 x 10 -8 less than about 10 nM (10 -8 Less than 5 nM (5 x 10 -9less than 1 nM (10 -9 less than about 0.1 nM (10 -10 less than about 0.01 nM (10 -11 M) or even lower than the equilibrium dissociation constant (K D ), where binding to a predetermined antigen occurs with an affinity that is at least two-fold higher than its affinity for binding to a nonspecific antigen other than the predetermined antigen or a closely related antigen (e.g., BSA, casein). The phrases "antibody recognizing an antigen" and "antibody specific for an antigen" are used interchangeably herein with the term "antibody that specifically binds to an antigen."

[0151] As used herein, the term "STAT1 phosphorylation" refers to the phosphorylation of the signal transduction and activator of transcription 1 (STAT1) polypeptide, a transcription factor encoded by the human STAT1 gene. STAT molecules are phosphorylated by receptor-associated kinases, leading to activation and dimerization by forming homodimers or heterodimers, which then translocate to the nucleus and act as transcription factors. STAT1 can be activated (phosphorylated) in response to signal transduction via several ligands, including IL-27. IL-27 signaling via IL-27R results in the phosphorylation of STAT1 (pSTAT1). STAT1 plays an important role in the expression of genes involved in cell survival, viability, or pathogen response. Methods for determining STAT1 phosphorylation as a result of IL-27 signaling include, but are not limited to, flow cytometry analysis of cells labeled with an antibody that specifically recognizes phosphorylated STAT1 (see, for example, Tochizawa et al., (2006) J Immunol Methods 313(1-2):29-37).

[0152] As used herein, the term "STAT3 phosphorylation" refers to the phosphorylation of signal transduction and activator of transcription 3 (STAT3) polypeptide, a transcription factor encoded by the human STAT3 gene. STAT3 mediates the expression of various genes in response to cell stimulation and plays an important role in many cellular processes, such as cell proliferation and apoptosis. Methods for determining STAT3 phosphorylation as a result of IL-27 signal transduction include, but are not limited to, the analysis of cells or cell extracts labeled with an antibody that specifically recognizes phosphorylated STAT3 (see, for example, Fursov et al., (2011) Assay Drug Dev Technol 9(4):420-429).

[0153] As used herein, the term "subject" includes any human or non-human animal. For example, the methods and compositions of the present disclosure can be used to treat subjects with immune disorders. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, including non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0154] With respect to nucleic acids, the term "substantial homology" indicates that at least about 80% of the nucleotides, usually at least about 90% to 95%, and more preferably at least about 98% to 99.5% of the nucleotides are identical, with appropriate nucleotide insertions or deletions, when two nucleic acids or their designated sequences are optimally aligned and compared. Alternatively, substantial homology exists when the segments will hybridize under selective hybridization conditions to the complement of the strand.

[0155] The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap (i.e., % homology = # of identical positions / total # of positions x 100). The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0156] The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at http: / / www.gcg.com) using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80, and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)) as incorporated into the ALIGN program (version 2.0), using a weight residue table of PAM120, a gap length penalty of 12, and a gap penalty of 4. Furthermore, percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm incorporated into the GAP program of the GCG software package (available at http: / / www.gcg.com) using either a Blossum62 matrix or a PAM250 matrix with gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6.

[0157] The nucleic acid and protein sequences of the present disclosure can also be used as "query sequences" to perform searches against public databases, for example, to identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the XBLAST program, score = 50, word length = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules of the present disclosure. BLAST protein searches can be performed with the XBLAST program, score = 50, word length = 3, to obtain amino acid sequences homologous to the protein molecules of the present disclosure. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov.

[0158] Nucleic acids can be present in whole cells, in cell lysates, or in partially purified or substantially pure form. Nucleic acids are "isolated" or "substantially purified" when they have been purified from other cellular components or other contaminants, such as other cellular nucleic acids or proteins, by standard techniques, including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art. See F. Ausubel, et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0159] The nucleic acid compositions of the present disclosure are often native sequences (except for modified restriction sites, etc.), but may be mutated according to standard techniques for providing gene sequences from either cDNA, genomes, or mixtures thereof. For coding sequences, these natural mutations may affect the amino acid sequence as desired. In particular, DNA sequences that are substantially homologous to or derived from the native V, D, J, constant, switch, and other such sequences described herein are contemplated (where "derived" indicates that the sequence is identical to or modified from another sequence).

[0160] As used herein, the term "STING" (or TMEM173) refers to stimulator of interferon genes, a protein that functions both as a direct cytoplasmic DNA sensor and as an adaptor protein. In humans, STING is encoded by the TMEM173 gene. STING plays an important role in innate immunity. STING induces type I interferon production when cells are infected with intracellular pathogens such as viruses, mycobacteria, and intracellular parasites. STING-mediated type I interferon protects infected cells and neighboring cells from local infection by binding to the same cell that secretes it and neighboring cells. An exemplary amino acid sequence of STING is provided by the NCBI Genbank database under accession number NP_001288667.

[0161] The term "T cell" refers to a type of white blood cell that can be distinguished from other white blood cells by the presence of a T cell receptor on the cell surface. There are several subsets of T cells, including, but not limited to, helper cells (also known as T H cells or CD4 + T cells) and T H 1. T H 2. T H 3. T H 17, T H 9. T FH A subtype containing cytotoxic T cells (also known as TC cells, CD8 + T cells, cytotoxic T lymphocytes, T killer cells, killer T cells), memory T cells and central memory T cells (T CM cells), effector memory T cells (T EM and T EMRA cells), and resident memory T cells (T RM subtypes, including regulatory T cells (also known as T reg cells or suppressor T cells) and CD4 + FOXP3 + T reg cells, CD4 + FOXP3 - T reg cells, Tr1 cells, Th3 cells, and T reg Subtypes include γδ T cells, including Vγ9 / Vδ2 T cells, natural killer T cells (also known as NKT cells), mucosal-associated invariant T cells (MAIT), and gamma delta T cells (γδ T cells), including Vγ9 / Vδ2 T cells. Any one or more of the foregoing T cells or T cells not mentioned may be the target cell type for the methods of use of the present disclosure.

[0162] As used herein, the term "T cell-mediated response" includes, but is not limited to, effector T cells (e.g., CD8 + cells) and helper T cells (e.g., CD4 + T cell-mediated responses include, for example, T cell cytotoxicity and proliferation.

[0163] As used herein, the term "therapeutically effective amount" or "therapeutically effective dose," or similar terms used herein, is intended to mean an amount of an agent (e.g., an anti-IL-27 antibody or antigen-binding fragment thereof) that elicits a desired biological or medical response (e.g., amelioration of one or more symptoms of cancer).

[0164] As used herein, the term "TAM receptor" refers to TAM receptor protein tyrosine kinases (TYRO3, AXL, and MER). TAM receptors are involved in controlling the homeostasis of the immune system. In the context of cancer, TAM receptors have a dual regulatory role: regulating the initiation and progression of tumor development while regulating the associated anti-tumor response of various immune cells. Further description of TAM receptors can be found in Paolino and Penninger (2016) Cancers 8(97): doi:10.3390 / cancers8100097). As used herein, the term "TAM receptor inhibitor" or "TAM inhibitor" refers to an agent that inhibits, blocks, or reduces the function or activity of TAM receptors.

[0165] As used herein, the term "TIGIT" or "T cell immunoreceptor having Ig and ITIM domains" refers to any native TIGIT from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. TIGIT is also known in the art as DKFZp667A205, FLJ39873, V-set and immunoglobulin domain-containing protein 9, V-set and transmembrane domain-containing protein 3, VSIG9, VSTM3, and WUCAM. This term also encompasses natural variants of TIGIT, such as splice variants or allelic variants. The amino acid sequence of an exemplary human TIGIT can be found under UniProt accession number Q495A1.

[0166] As used herein, the terms "treat," "treating," and "treatment" refer to therapeutic or preventative measures as described herein. The method of "treatment" involves administering a human antibody of the present disclosure to a subject in need of such treatment, e.g., a subject in need of an enhanced immune response to a particular antigen, or a subject who may ultimately suffer from such a disorder, to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of the disorder or recurring disorder, or prolong the subject's survival beyond that expected in the absence of such treatment.

[0167] As used herein, the term "tumor microenvironment" (or "cancer microenvironment," abbreviated TME) refers to the cellular milieu or environment in which a tumor or neoplasm resides, including surrounding blood vessels and non-cancerous cells, including, but not limited to, immune cells, fibroblasts, bone marrow-derived inflammatory cells, and lymphocytes. Signaling molecules and the extracellular matrix also comprise the TME. Tumors and the surrounding microenvironment are closely associated and constantly interact. Tumors can influence the microenvironment by releasing extracellular signals, promoting tumor angiogenesis, and inducing peripheral immune tolerance, while immune cells within the microenvironment can influence the growth and evolution of tumor cells.

[0168] As used herein, the term "tumor proportion score" (TPS) refers to the number of positive tumor cells divided by the total number of viable tumor cells multiplied by 100%.

[0169] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, in which additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Furthermore, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. As used herein, "plasmid" and "vector" may be used interchangeably, as the plasmid is the most commonly used form of vector. However, the present disclosure is intended to include such other forms of expression vectors, such as viral vectors (eg, replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

[0170] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as that commonly understood by those skilled in the art in the field to which this disclosure pertains.Preferred methods and materials are described below, but similar or equivalent methods and materials to those described herein can also be used in the implementation or testing of the method and composition currently disclosed.All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety.

[0171] II. Methods of the Disclosure Some embodiments of the present disclosure relate to methods of identifying subjects suitable for treatment comprising an IL-27 inhibitor, e.g., an anti-IL-27 antibody disclosed herein. The present disclosure surprisingly found that expression of various biomarkers can indicate a patient's responsiveness to an IL-27 inhibitor, e.g., an anti-IL-27 antibody disclosed herein. In some embodiments, a patient suitable for treatment comprising an IL-27 inhibitor, e.g., an anti-IL-27 antibody disclosed herein, is identified as having a tumor comprising tumor cells that express WSX-1, referred to herein as a "WSX-1-positive tumor." In some embodiments, a patient suitable for treatment comprising an IL-27 inhibitor, e.g., an anti-IL-27 antibody disclosed herein, is identified as having a tumor comprising one or more immune cells that express IL-27 within the tumor.

[0172] A. WSX-1 positive tumor Some embodiments of the present disclosure relate to a method for treating a tumor in a subject, comprising administering an IL-27 inhibitor to the subject, wherein the tumor is identified as a WSX-1-positive tumor. Some embodiments of the present disclosure relate to a method for treating a tumor in a subject in need of treatment, comprising: (i) identifying a subject with a WSX-1-positive tumor; and (ii) administering an IL-27 inhibitor to the subject. In some embodiments, the WSX-1-positive tumor is identified by detecting WSX-1 expression in a tumor sample obtained from the subject.

[0173] Some embodiments of the present disclosure relate to a method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, the method comprising detecting WSX-1 in a tumor sample obtained from the subject. In some embodiments, the method further comprises administering an IL-27 inhibitor to the subject identified as having a WSX-1-positive tumor.

[0174] In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy. In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy. In some embodiments, the tumor sample obtained from the subject comprises tumor cells, tumor-infiltrating lymphocytes, or both. In some embodiments, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of the cells in the tumor sample express WSX-1. In some embodiments, at least about 1% of the cells in the tumor sample express WSX-1. In some embodiments, at least about 2% of the cells in the tumor sample express WSX-1. In some embodiments, at least about 3% of the cells in the tumor sample express WSX-1. In some embodiments, at least about 4% of the cells in the tumor sample express WSX-1. In some embodiments, at least about 5% of cells in the tumor sample express WSX-1. In some embodiments, at least about 10% of cells in the tumor sample express WSX-1. In some embodiments, at least about 15% of cells in the tumor sample express WSX-1. In some embodiments, at least about 20% of cells in the tumor sample express WSX-1. In some embodiments, at least about 25% of cells in the tumor sample express WSX-1. In some embodiments, at least about 30% of cells in the tumor sample express WSX-1. In some embodiments, at least about 35% of cells in the tumor sample express WSX-1. In some embodiments, at least about 40% of cells in the tumor sample express WSX-1. In some embodiments, at least about 45% of cells in the tumor sample express WSX-1. In some embodiments, at least about 50% of cells in the tumor sample express WSX-1. In some embodiments, at least about 60% of cells in the tumor sample express WSX-1.In some embodiments, at least about 70% of cells in a tumor sample express WSX-1. In some embodiments, at least about 80% of cells in a tumor sample express WSX-1. In some embodiments, at least about 90% of cells in a tumor sample express WSX-1. In some embodiments, cells in a tumor sample that express WSX-1 are tumor cells.

[0175] Any means for detecting WSX-1 expression can be used in the methods disclosed herein. In some embodiments, WSX-1 protein expression is detected. In some embodiments, WSX-1 expression is detected using an immunohistochemistry (IHC) assay. In some embodiments, WSX-1 expression is detected using an automated IHC assay. In some embodiments, WSX-1 expression is detected by contacting a tumor sample with an antibody or antigen-binding portion thereof that specifically binds to human WSX-1. In some embodiments, expression of RNA encoding WSX-1 is detected.

[0176] In some embodiments, a tumor sample obtained from a subject is characterized by WSX-1 expression measured by an assay (e.g., an immunohistochemistry (IHC) assay) of 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) measured by an assay of 1% or more. In some embodiments, a sample is characterized by a tumor proportion score (TPS) measured by an assay of 5% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) measured by an assay of 10% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) measured by an assay of 25% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) measured by an assay of 50% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a TPS of 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more WSX-1 expression as measured by an assay (e.g., an immunohistochemistry (IHC) assay). In some embodiments, WSX-1 expression is assessed using a tumor proportion score. In some embodiments, the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50%, or at least 60%. In some embodiments, the TPS or CPS is at least 10%. In some embodiments, the TPS or CPS is at least 20%. In some embodiments, the TPS or CPS is at least 30%. In some embodiments, the TPS or CPS is at least 40%. In some embodiments, the TPS or CPS is at least 50%. In some embodiments, the TPS or CPS is at least 60%. In some embodiments, the TPS is higher than the CPS for the same sample.In some embodiments, TPS or CPS correlates with response to treatment with an IL-27 inhibitor.

[0177] The percentage of cells that express WSX-1 in tumor sample can be determined by any means.In some embodiments, IHC assay is used to mark all WSX-1-expressing cells in tumor sample, then count the total cells in tumor sample (for example, PPFE sample), then count the cells that express WSX-1, and divide the number of cells that express WSX-1 by the total number of cells in tumor sample, and multiply by 100.In some embodiments, the percentage of cells that express WSX-1 is averaged over multiple PPFE samples that are prepared from a single tumor sample (for example, tumor biopsy).

[0178] B.IL-27 positive immune cells Some embodiments of the present disclosure relate to a method for treating a tumor in a subject, the method comprising administering an IL-27 inhibitor to the subject, wherein one or more immune cells in a tumor sample obtained from the subject express IL-27. Some embodiments of the present disclosure relate to a method for treating a tumor in a subject in need thereof, the method comprising: (i) identifying a subject having a tumor in which one or more immune cells in a tumor sample obtained from the subject express IL-27; and (ii) administering an IL-27 inhibitor to the subject.

[0179] Some embodiments of the present disclosure relate to a method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, the method comprising detecting IL-27 expression in a tumor sample obtained from the subject. In some embodiments, the method further comprises administering an IL-27 inhibitor to a subject identified as having a tumor sample comprising one or more immune cells that express IL-27.

[0180] In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy. In some embodiments, the tumor sample obtained from the subject is a tumor tissue biopsy. In some embodiments, the tumor sample obtained from the subject comprises tumor cells, tumor-infiltrating lymphocytes, or both. One or more immune cells in the tumor sample comprise macrophages.

[0181] In some embodiments, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 1% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 2% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 3% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 4% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 5% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 10% of immune cells in a tumor sample express IL-27. In some embodiments, at least about 15% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 20% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 25% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 30% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 35% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 40% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 45% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 50% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 60% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 70% of immune cells in the tumor sample express IL-27. In some embodiments, at least about 80% of immune cells in the tumor sample express IL-27.In some embodiments, at least about 90% of immune cells in the tumor sample express IL-27.

[0182] Any means of detecting IL-27 expression can be used in the methods disclosed herein. In some embodiments, expression of IL-27 protein is detected. In some embodiments, IL-27 expression is detected using an immunohistochemistry (IHC) assay. In some embodiments, IL-27 expression is detected using an automated IHC assay. In some embodiments, v expression is detected by contacting a tumor sample with an antibody or antigen-binding portion thereof that specifically binds to human IL-27. In some embodiments, expression of RNA encoding IL-27 is detected.

[0183] The percentage of immune cells expressing IL-27 in tumor samples can be determined by any means.In some embodiments, IHC assay is used to label all IL-27-expressing cells in tumor samples, and then the total number of immune cells in tumor samples (e.g., PPFE samples) is counted, and then the cells expressing IL-27 are counted, and the number of cells expressing IL-27 is divided by the total number of immune cells in tumor samples and multiplied by 100.In some embodiments, the percentage of immune cells expressing IL-27 is the average of multiple PPFE samples prepared from a single tumor sample (e.g., tumor biopsy).

[0184] In some embodiments, a tumor sample obtained from a subject is characterized by IL-27 expression as measured by an assay (e.g., an immunohistochemistry (IHC) assay) of 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) as measured by an assay of 1% or more. In some embodiments, the sample is characterized by a tumor proportion score (TPS) as measured by an assay of 5% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) as measured by an assay of 10% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) as measured by an assay of 25% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a tumor proportion score (TPS) as measured by an assay of 50% or more. In some embodiments, a tumor sample obtained from a subject is characterized by a TPS of 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more IL-27 expression as measured by an assay (e.g., an immunohistochemistry (IHC) assay). In some embodiments, IL-27 expression is assessed using a tumor proportion score. In some embodiments, the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50%, or at least 60%. In some embodiments, the TPS or CPS is at least 10%. In some embodiments, the TPS or CPS is at least 20%. In some embodiments, the TPS or CPS is at least 30%. In some embodiments, the TPS or CPS is at least 40%. In some embodiments, the TPS or CPS is at least 50%. In some embodiments, the TPS or CPS is at least 60%. In some embodiments, the TPS is higher than the CPS for the same sample.In some embodiments, TPS or CPS correlates with response to treatment with an IL-27 inhibitor.

[0185] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.003 mg / kg to at least about 20 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to an epitope comprising one or more amino acids of (i) amino acids 37 to 56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142 to 164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii). In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose sufficient to maintain an IC90 of pSTAT1 inhibition level, i.e., greater than about 0.7 μg / mL, for the duration of treatment, e.g., 28 days, 56 days, or 84 days.

[0186] In some embodiments, the antibody or antigen-binding portion thereof is at least about 0.006 mg / kg to at least about 20 mg / kg, at least about 0.009 mg / kg to at least about 20 mg / kg, at least about 0.01 mg / kg to at least about 20 mg / kg, at least about 0.03 mg / kg to at least about 20 mg / kg, at least about 0.06 mg / kg to at least about 20 mg / kg, at least about 0.09 mg / kg to at least about 20 mg / kg, at least about 0.1 mg / kg to at least about 20 mg / kg, at least about 0.3 mg / kg to at least about 20 mg / kg, at least about 0.6 mg / kg to at least about 20 mg / kg, at least about 0.9 mg / kg to at least about 20 mg / kg, at least about 1 mg / kg to at least about 20 mg / kg, at least about 1 mg / kg to at least about 20 mg / kg, / kg to at least about 20 mg / kg, at least about 3 mg / kg to at least about 20 mg / kg, at least about 6 mg / kg to at least about 20 mg / kg, at least about 10 mg / kg to at least about 20 mg / kg, at least about 13 mg / kg to at least about 20 mg / kg, at least about 13 mg / kg to at least about 18 mg / kg, at least about 13 mg / kg to at least about 16 mg / kg, at least about 16 mg / kg to at least about 20 mg / kg, at least about 16 mg / kg to at least about 18 mg / kg, at least about 3 mg / kg to at least about 18 mg / kg, at least about 6 mg / kg to at least about 15 mg / kg, at least about 13 mg / kg to at least about 18 mg / kg, or at least about 10 mg / kg to at least about 15 mg / kg.In some embodiments, the antibody or antigen-binding portion thereof is at least about 0.006 mg / kg to at least about 10 mg / kg, at least about 0.009 mg / kg to at least about 10 mg / kg, at least about 0.01 mg / kg to at least about 10 mg / kg, at least about 0.03 mg / kg to at least about 10 mg / kg, at least about 0.06 mg / kg to at least about 10 mg / kg, at least about 0.09 mg / kg to at least about 10 mg / kg, at least about 0.1 mg / kg to at least about 10 mg / kg, at least about The dose may be administered at a dose of 0.3 mg / kg to at least about 10 mg / kg, at least about 0.6 mg / kg to at least about 10 mg / kg, at least about 0.9 mg / kg to at least about 10 mg / kg, at least about 1 mg / kg to at least about 10 mg / kg, at least about 1 mg / kg to at least about 9 mg / kg, at least about 3 to at least about 9 mg / kg, at least about 1 mg / kg to at least about 6 mg / kg, at least about 3 mg / kg to at least about 6 mg / kg, or at least about 1 mg / kg to at least about 3 mg / kg.

[0187] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.003 mg / kg, at least about 0.006 mg / kg, at least about 0.009 mg / kg, at least about 0.01 mg / kg, at least about 0.03 mg / kg, at least about 0.06 mg / kg, at least about 0.09 mg / kg, at least about 0.1 mg / kg, at least about 0.3 mg / kg, at least about 0.6 mg / kg, at least about 0.9 mg / kg, at least about 1.0 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, or at least about 10 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, at least about 18 mg / kg, at least about 19 mg / kg, or at least about 20 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.003 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.006 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.009 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.01 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.03 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.06 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.09 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.1 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.3 mg / kg.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.6 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 0.9 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 1.0 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 2 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 3 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 4 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 5 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 6 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 7 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 8 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 9 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 10 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 11 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 12 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 13 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 14 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 15 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 16 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 17 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 18 mg / kg.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 19 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of at least about 20 mg / kg.

[0188] In some embodiments, the antibody or antigen-binding portion thereof is administered about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every six weeks, about once every eight weeks, or about once every twelve weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered about once every four weeks.

[0189] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 0.3 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 1 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 2 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 3 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 4 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 5 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 6 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 7 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 8 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 9 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 11 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 12 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 13 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 14 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 15 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 16 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 17 mg / kg about once per week.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 18 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 19 mg / kg about once per week. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 20 mg / kg about once per week.

[0190] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 0.3 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 1 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 2 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 3 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 4 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 5 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 6 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 7 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 8 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 9 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 11 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 12 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 13 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 14 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 15 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 16 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 17 mg / kg about once every two weeks.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 18 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 19 mg / kg about once every two weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 20 mg / kg about once every two weeks.

[0191] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 0.3 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 1 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 2 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 3 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 4 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 5 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 6 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 7 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 8 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 9 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 11 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 12 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 13 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 14 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 15 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 16 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 17 mg / kg about once every three weeks.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 18 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 19 mg / kg about once every three weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 20 mg / kg about once every three weeks.

[0192] In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 0.3 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 1 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 2 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 3 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 4 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 5 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 6 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 7 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 8 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 9 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 10 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 11 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 12 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 13 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 14 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 15 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 16 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 17 mg / kg about once every four weeks.In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 18 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 19 mg / kg about once every four weeks. In some embodiments, the antibody or antigen-binding portion thereof is administered at a dose of about 20 mg / kg about once every four weeks.

[0193] Certain aspects of the present disclosure relate to methods of treating cancer in a subject in need thereof. In some embodiments, the cancer is selected from the group consisting of Kaposi's sarcoma, leukemia, acute lymphocytic leukemia, acute myeloid leukemia, myeloblastic promyelocytic monocytic erythroleukemia, chronic leukemia, chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt's lymphoma and marginal zone B-cell lymphoma, polycythemia vera lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, solid tumors, sarcomas, and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic s ... sarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colorectal sarcoma, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma (RCC), hepatocellular carcinoma (HCC), hepatoma, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular tumor, lung cancer, small cell lung cancer, non-small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma , meningioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal cancer, esophageal cancer, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer, intraepithelial neoplasia, kidney cancer, laryngeal cancer, liver cancer, lung cancer (small cell carcinoma, large cell carcinoma), melanoma, neuroblastoma, oral cancer (e.g., lip, tongue, oral cavity, and pharynx), ovarian cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, cancer of the respiratory system, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, cancer of the urinary system, and any combination thereof.In some embodiments, the cancer is selected from lung cancer (e.g., non-small cell lung cancer), sarcoma, testicular cancer, ovarian cancer, pancreatic cancer, breast cancer (e.g., triple-negative breast cancer), melanoma, head and neck cancer (e.g., head and neck squamous cell carcinoma), colon cancer, bladder cancer, endometrial cancer, prostate cancer, thyroid cancer, hepatocellular carcinoma, gastric cancer, brain tumor, lymphoma (e.g., DL-BCL), leukemia (e.g., AML), or kidney cancer (e.g., renal cell carcinoma, e.g., clear cell RCC and / or non-clear cell RCC). In some embodiments, the method can be performed in combination with other cancer treatments. For example, the composition can be administered to a subject simultaneously with, before, or after radiation therapy, surgery, targeted chemotherapy or cytotoxic chemotherapy, chemoradiotherapy, hormone therapy, immunotherapy, gene therapy, cell transplantation therapy, precision medicine, genome editing therapy, or other drug therapy.

[0194] In some embodiments, the compositions disclosed herein are administered to a subject, e.g., a human subject, using a variety of methods, depending in part on the route of administration. The route can be, for example, intravenous injection or infusion (IV), subcutaneous injection (SC), intraperitoneal (IP) injection, intramuscular injection (IM), or intrathecal injection (IT). The injection can be a bolus or continuous infusion.

[0195] Administration can be achieved, for example, by local infusion, injection, or implant. The implant can be a porous, non-porous, or gelatinous material, including a membrane such as a silastic membrane or a fiber. The implant can be configured to continuously or periodically release the composition to the subject. See, for example, U.S. Patent Application Publication No. 20080241223; U.S. Patent Nos. 5,501,856 and 3,710,795, EP488401, and EP430539. The disclosures of each of these are incorporated herein by reference in their entirety. The composition can be delivered to the subject by, for example, a diffusion system, an erodible system, or a convective system, such as an osmotic pump, a biodegradable implant, an electrodiffusion system, an electroosmotic system, a vapor pressure pump, an electrolytic pump, an effervescent pump, a piezoelectric pump, an erosion-based system, or an implantable device based on an electromechanical system.

[0196] In some embodiments, the anti-IL-27 antibody or antigen-binding fragment thereof is therapeutically delivered to the subject by local administration.

[0197] In certain embodiments, the administration route follows known methods, such as oral, intravenous, intraperitoneal, intracerebral (intraparenchymal), intracerebroventricular, intramuscular, subcutaneous, intraocular, intraarterial, intraportal or intralesional injection; by sustained release system or by implantation device.In certain embodiments, the composition can be administered by bolus injection, or continuously by infusion, or by implantation device.In certain embodiments, the individual components of the combination therapy can be administered by different routes.

[0198] In certain embodiments, the composition may be administered locally via implantation of a membrane, sponge, or other suitable material into which the desired molecule has been absorbed or encapsulated. In certain embodiments, when an implantation device is used, the device may be implanted into any suitable tissue or organ, and delivery of the desired molecule may be by diffusion, sustained bolus, or continuous administration. In certain embodiments, it may be desirable to use a pharmaceutical composition comprising an anti-IL-27 antibody in an ex vivo method. In such cases, cells, tissues, and / or organs removed from a patient are exposed to a pharmaceutical composition comprising an anti-IL-27 antibody, and then the cells, tissues, and / or organs are subsequently transplanted back into the patient.

[0199] In certain embodiments, anti-IL-27 antibodies can be delivered by implanting certain cells that have been genetically engineered using methods such as those described herein to express and secrete the polypeptide. In certain embodiments, such cells can be animal or human cells, and can be autologous, heterologous, or xenogeneic. In certain embodiments, the cells can be immortalized. In certain embodiments, to reduce the likelihood of an immune response, the cells can be encapsulated to avoid infiltration of surrounding tissues. In certain embodiments, the encapsulating material is typically a biocompatible, semipermeable polymeric enclosure or membrane, which can release the protein product(s) but prevent destruction of the cells by the patient's immune system or other harmful factors from surrounding tissues.

[0200] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of one or more biomarkers selected from the group consisting of eotaxin-1 (CCL11), TARC (CCL17), VEGF-A, IL-7, IL-8, MCP-1, MCP-4, and any combination thereof, wherein the increased expression of the one or more biomarkers is relative to the expression of the one or more biomarkers prior to administration. In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of eotaxin-1 (CCL11), wherein the increased expression of eotaxin-1 (CCL11) is relative to the expression of eotaxin-1 (CCL11) prior to administration.

[0201] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of TARC (CCL17), wherein the increased expression of TARC (CCL17) is relative to the expression of TARC (CCL17) prior to administration.

[0202] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of VEGF-A, wherein the increased expression of VEGF-A is relative to the expression of VEGF-A prior to administration.

[0203] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of IL-7, wherein the increased expression of IL-7 is relative to the expression of IL-7 before administration.

[0204] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of IL-8, wherein the increased expression of IL-8 is relative to the expression of IL-8 before administration.

[0205] In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of MCP-1, wherein the increased expression of MCP-1 is relative to the expression of MCP-1 prior to administration. In some embodiments, after administration of the antibody or antigen-binding portion thereof, the subject exhibits increased expression of MCP-4, wherein the increased expression of MCP-4 is relative to the expression of MCP-4 prior to administration.

[0206] C. IL-27 inhibitors Some embodiments of the present disclosure relate to methods of administering an IL-27 inhibitor to a subject, wherein the subject is identified as suitable for IL-27 inhibitor treatment. Any molecule capable of inhibiting the activity of IL-27 or its receptor (WSX-1) can be used in the methods disclosed herein. In some embodiments, the IL-27 inhibitor reduces or blocks the interaction between IL-27 and WSX-1. In some embodiments, the inhibitor reduces or blocks downstream IL-27 signaling.

[0207] In some embodiments, the IL-27 inhibitor comprises a polypeptide. In some embodiments, the IL-27 inhibitor comprises a small molecule. In some embodiments, the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27 (an "anti-IL-27 antibody"). In some embodiments, the antibody or antigen-binding portion thereof specifically binds to IL-27p28 and antagonizes IL-27, particularly human IL-27.

[0208] In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces phosphorylation of STAT1 and / or STAT3 in a cell of the subject. In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces pSTAT1 signaling (e.g., IL-27-mediated pSTAT1 signaling). In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces inhibition of CD161 expression in a cell of the subject. In some embodiments, the antibody or antigen-binding portion thereof inhibits or reduces PD-L1 expression in a cell of the subject. In some embodiments, the antibody or antigen-binding portion thereof induces or enhances PD-1-mediated secretion of one or more cytokines from a cell of the subject. In some embodiments, the antibody or antigen-binding portion thereof alters TIM-3 expression in a cell of the subject. In some embodiments, the cell is a tumor cell or an immune cell.

[0209] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to an epitope comprising one or more amino acids of (i) amino acids 37 to 56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142 to 164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii). In some embodiments, an isolated antibody of the present disclosure, or antigen-binding portion thereof, that antagonizes human IL-27 specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, or Glu164 of SEQ ID NO: 2 (IL-27p28).

[0210] In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR3 comprising the sequence set forth in SEQ ID NO: 121. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR3 comprising the sequence set forth in SEQ ID NO: 124. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR2 comprising the sequence set forth in SEQ ID NO: 120 or 123. In some embodiments, the antibody, or portion thereof, comprises a heavy chain CDR2 comprising the sequence set forth in SEQ ID NO: 120. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR2 comprising the sequence set forth in SEQ ID NO: 123. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR1 comprising the sequence set forth in SEQ ID NO: 119 or 122. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR1 comprising the sequence set forth in SEQ ID NO: 119. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR1 comprising the sequence set forth in SEQ ID NO: 122. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a light chain CDR3 comprising the sequence set forth in SEQ ID NO: 129 or 132. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a light chain CDR3 comprising the sequence set forth in SEQ ID NO: 129. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR3 comprising the sequence set forth in SEQ ID NO: 132. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR2 comprising the sequence set forth in SEQ ID NO: 128 or 131. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR2 comprising the sequence set forth in SEQ ID NO: 128. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR2 comprising the sequence set forth in SEQ ID NO: 131. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the sequence set forth in SEQ ID NO: 127 or 130. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the sequence set forth in SEQ ID NO: 127. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the sequence set forth in SEQ ID NO: 130.

[0211] In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 122, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124.

[0212] In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 132.

[0213] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121, a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129.

[0214] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 125. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 125.

[0215] In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 133. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 133.

[0216] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 135. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 135.

[0217] In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 139. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 139.

[0218] In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 137. In some embodiments, the antibody or antigen-binding portion thereof comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 137.

[0219] In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 0.003 mg / kg to at least about 20 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 1 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 2 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 3 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 4 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 5 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 6 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 7 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 8 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 9 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 10 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 11 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 12 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 13 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 14 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 15 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 16 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 17 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 18 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 19 mg / kg. In some embodiments, the anti-IL-27 antibody is administered at a dose of at least about 20 mg / kg.

[0220] In some embodiments, the antibody or antigen-binding portion thereof comprises an amino acid sequence set forth in Table 1A.

[0221] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14] [Table 1-15]

[0222] In some embodiments, the antibody, or antigen-binding portion thereof, comprises an Fc sequence set forth in Table 1B. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain, wherein the heavy chain comprises an Fc region having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to the sequence set forth in SEQ ID NO:5, 6, 7, or 8. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain, wherein the heavy chain comprises an Fc region comprising the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain, wherein the heavy chain comprises an Fc region comprising the amino acid sequence set forth in SEQ ID NO:6. In some embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain, wherein the heavy chain comprises an Fc region comprising the amino acid sequence set forth in SEQ ID NO:7. Some antibodies, or antigen-binding portion thereof, comprise a heavy chain, wherein the heavy chain comprises an Fc region comprising the amino acid sequence set forth in SEQ ID NO:8.

[0223] [Table 2]

[0224] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Asp146 and Arg149 of SEQ ID NO:2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Asp146 and Phe153 of SEQ ID NO:2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Arg149 and Phe153 of SEQ ID NO:2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Asp146, Arg149, and / or Phe153 of SEQ ID NO:2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Asp146, Arg149, and / or Phe153 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope comprises Asp146, Arg149, His150, and Phe153 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope comprises Asp146, Arg149, Phe153, and Leu156 of SEQ ID NO:2 (IL-27p28). In some embodiments, the epitope comprises Asp146, Arg149, His150, Phe153, and Leu156 of SEQ ID NO:2 (IL-27p28).

[0225] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising at least one, at least two, at least three, at least four, at least five, or at least six amino acids of IL-27p28 selected from Leu142, Asp146, Arg149, His150, Phe153, Leu156, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Leu142, Asp146, Arg149, His150, Phe153, Leu156, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, the epitope comprises Gln37, Leu38, Glu42, Asp146, Arg149, His150, Phe153, and Leu156 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Gln37, Leu38, Glu42, Leu142, Asp146, Arg149, His150, Phe153, Leu156, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0226] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising at least one, at least two, at least three, at least four, at least five, or at least six, at least seven, at least eight, or at least nine amino acids of IL-27p28 selected from Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phel53, Leu156, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising at least one, at least two, at least three, at least four, at least five, or at least six, at least seven, at least eight, or at least nine amino acids of IL-27p28 selected from Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phel53, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28). In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope including Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0227] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope consisting of or consisting essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0228] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising at least one residue selected from the group consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), and Leu53, Lys56, Asp143, Leu147, Arg152, Ala157, Gly159, Phe160, or Asn161 of SEQ ID NO: 2 (IL-27p28).

[0229] In some embodiments, an antibody or antigen-binding portion thereof of the present disclosure specifically binds to an epitope comprising at least one residue selected from the group consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), and Leu53, Lys56, Asp143, Arg145, Leu147, Arg152, Ala157, Gly159, Phe160, Asn161, or Pro163 of SEQ ID NO: 2 (IL-27p28).

[0230] In some embodiments, an antibody of the disclosure, or an antigen-binding portion thereof, specifically binds to an epitope consisting of or consisting essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0231] In some embodiments, an antibody or antigen-binding portion thereof of the disclosure specifically binds to an epitope consisting of or consisting essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0232] In some embodiments, the disclosure provides antibodies or antigen-binding portions thereof that specifically bind to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), and have the following properties: (i) an equilibrium dissociation constant (K D(ii) binds to human IL-27 at a specific site; (ii) blocks the binding of IL-27 to the IL-27 receptor; (iii) inhibits or reduces phosphorylation of STAT1 and / or STAT3 in a cell; (iv) inhibits or reduces the inhibition of CD161 expression in a cell; (v) inhibits or reduces PD-L1 expression in a cell; (vi) induces or enhances PD-1-mediated secretion of one or more cytokines from a cell; (vii) alters TIM-3 expression in a cell; and (viii) a combination of (i)-(vii).

[0233] In some embodiments, the isolated antibody or antigen-binding portion thereof binds to an epitope of one or more amino acids selected from Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (human IL-27p28) with an equilibrium dissociation constant (K D ) to join them.

[0234] In some embodiments, the isolated antibody or antigen-binding portion thereof binds to recombinant human IL-27p28. In some embodiments, the isolated antibody or antigen-binding portion thereof binds to murine IL-27p28.

[0235] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in cells. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in cells. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 and STAT3 phosphorylation in cells. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in a cell by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% compared to STAT1 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in a cell by at least about 50% compared to STAT1 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in a cell by at least about 60% compared to STAT1 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in a cell by at least about 70% compared to STAT1 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in a cell by at least about 75% compared to STAT1 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof.In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in cells by at least about 80% compared to STAT1 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in cells by at least about 85% compared to STAT1 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in cells by at least about 90% compared to STAT1 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT1 phosphorylation in cells by at least about 95% compared to STAT1 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof eliminates STAT1 phosphorylation in cells.

[0236] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 50%, compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 60% compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 70% compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 75% compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in a cell by at least about 80% compared to STAT3 phosphorylation in the cell before contacting the cell with the antibody or antigen-binding portion thereof.In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in cells by at least about 85% compared to STAT3 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in cells by at least about 90% compared to STAT3 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces STAT3 phosphorylation in cells by at least about 95% compared to STAT3 phosphorylation in the cells before contacting the cells with the antibody or antigen-binding portion thereof. In some embodiments, the isolated antibody or antigen-binding portion thereof eliminates STAT3 phosphorylation in cells.

[0237] In some embodiments, the cell is an immune cell. In some embodiments, the cell is a cancer cell.

[0238] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces inhibition of CD161 expression in the cell (e.g., ameliorates or alleviates inhibition of CD161 expression in the cell). In some embodiments, the cell is an immune cell.

[0239] In some embodiments, the isolated antibody or antigen-binding portion thereof inhibits or reduces PD-L1 expression in a cell. In some embodiments, PD-L1 expression is inhibited or reduced. In some embodiments, TIM-3 expression is altered. In some embodiments, both PD-L1 expression and TIM-3 expression are altered. In some embodiments, the cell is an immune cell. In some embodiments, the antibody is a monoclonal antibody.

[0240] In some embodiments, the isolated antibody or antigen-binding portion thereof induces or enhances PD-1-mediated secretion of one or more cytokines from a cell. In some embodiments, the one or more cytokines are TNFα. In some embodiments, the one or more cytokines are IL-6. In some embodiments, the one or more cytokines are TNFα and IL-6. In some embodiments, the cell is an immune cell.

[0241] In some embodiments, the isolated portion or antigen-binding portion thereof is selected from the group consisting of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE antibodies. In some embodiments, the antibody is an IgG1 antibody or an IgG4 antibody. In some embodiments, the antibody comprises a wild-type IgG1 heavy chain constant region. In some embodiments, the antibody comprises a wild-type IgG4 heavy chain constant region. In some embodiments, the antibody comprises an Fc domain comprising at least one mutation. In some embodiments, the antibody comprises a mutated IgG1 heavy chain constant region. In some embodiments, the antibody comprises a mutated IgG4 heavy chain constant region. In some embodiments, the naturally mutated IgG4 heavy chain constant region comprises any one of the substitutions S228P, L235E, L235A, or a combination thereof, according to EU numbering.

[0242] In some embodiments, the present disclosure provides an isolated antibody, or antigen-binding portion thereof, that binds to substantially the same epitope on IL-27 as an antibody, or antigen-binding portion thereof, according to any one of the preceding embodiments.

[0243] In some embodiments, the present disclosure provides an isolated antibody or antigen-binding portion thereof that binds to at least one amino acid residue selected from the group consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), which is bound by the antibody or antigen-binding portion thereof according to any one of the preceding embodiments.

[0244] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof, wherein a mutation of the epitope bound by the antibody or antigen-binding portion thereof (Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28)) inhibits, reduces, or blocks binding to both the antibody or antigen-binding portion thereof and an antibody or antigen-binding portion thereof according to any one of the preceding embodiments.

[0245] In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, where light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-C. In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, where light chain CDR3 consists of N-XXXASAXXX-C. In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, where heavy chain CDR2 consists of N-XXSSSXSYXYXXXXXXX-C. In some embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3, a light chain CDR1, a light chain CDR2, and a light chain CDR3, wherein the heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, where X is any amino acid.

[0246] In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, where light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-C and light chain CDR3 consists of N-XXXASAXXX-C. In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, where heavy chain CDR2 consists of N-XXXSSSXSYXYXXXXXXX-C and heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, where X is any amino acid.

[0247] In some embodiments, the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, wherein light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-C, light chain CDR3 consists of N-XXXASAXXX-C, heavy chain CDR2 consists of N-XXSSSXSYXYXXXXXXX-C, and heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, where X is any amino acid.

[0248] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 31, 32 and 33, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 39, 40 and 41, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 53, 54 and 55, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 61, 62 and 63, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 75, 76 and 77, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 83, 84 and 85, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 97, 98 and 99, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 105, 106 and 107, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 119, 120, and 121, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0249] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 31, 32 and 33, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 39, 40 and 41, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 53, 54 and 55, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 61, 62 and 63, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 75, 76 and 77, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 83, 84 and 85, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 97, 98 and 99, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 105, 106 and 107, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 119, 120, and 121, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0250] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 9, 10, and 11, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 17, 18, and 19, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 31, 32 and 33, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 39, 40 and 41, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 53, 54 and 55, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 61, 62 and 63, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 75, 76 and 77, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 83, 84 and 85, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 97, 98 and 99, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 105, 106 and 107, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 119, 120, and 121, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0251] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 12, 13, and 14, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 34, 35 and 36, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 42, 43 and 44, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 56, 57 and 58, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 64, 65 and 66, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 78, 79 and 80, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 86, 88 and 89, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 100, 101 and 102, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 108, 109 and 110, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 122, 123, and 124, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 130, 131, and 132, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0252] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 12, 13, and 14, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 34, 35 and 36, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 42, 43 and 44, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 56, 57 and 58, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 64, 65 and 66, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 78, 79 and 80, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 86, 88 and 89, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 100, 101 and 102, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 108, 109 and 110, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 122, 123, and 124, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 130, 131, and 132, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0253] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 12, 13, and 14, respectively; and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 20, 21, and 22, respectively; (ii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 34, 35 and 36, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 42, 43 and 44, respectively; (iii) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 56, 57 and 58, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 64, 65 and 66, respectively; (iv) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 78, 79 and 80, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 86, 88 and 89, respectively; (v) heavy chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 100, 101 and 102, respectively, and light chain CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 108, 109 and 110, respectively; or (vi) heavy chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 122, 123, and 124, respectively, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 130, 131, and 132, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not comprise a heavy chain CDR and a light chain CDR selected from the group consisting of:

[0254] In some embodiments, the present disclosure provides an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). Provided is an isolated antibody or antigen-binding portion thereof that specifically binds, wherein the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, and wherein heavy chain CDR1 does not consist of N-GFTF[S / A / R][S / R][T / Y][G / S]-C (SEQ ID NO: 144) and / or heavy chain CDR2 does not consist of N-ISSS[S / G][S / A]YI-C (SEQ ID NO: 146).

[0255] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof and wherein the heavy chain CDR1 does not consist of N-GFTF[S / A / R][S / R][T / Y][G / S]-C (SEQ ID NO: 144) and / or the heavy chain CDR2 does not consist of N-ISSS[S / G][S / A]YI-C (SEQ ID NO: 146).

[0256] In some embodiments, the present disclosure provides an antibody specifically targeting an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). Provided is an isolated antibody or antigen-binding portion thereof that binds, wherein the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, and wherein heavy chain CDR1 does not consist of N-GFTF[S / A / R][S / R][T / Y][G / S]-C (SEQ ID NO: 144) and / or heavy chain CDR2 does not consist of N-ISSS[S / G][S / A]YI-C (SEQ ID NO: 146).

[0257] In some embodiments, the present invention provides an antibody specifically targeting an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). An isolated antibody or antigen-binding portion thereof is provided, wherein the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, and wherein heavy chain CDR1 does not comprise N-FTF[S / A / R][S / R][T / Y][G / S]MN-C (SEQ ID NO: 148) and / or heavy chain CDR2 does not comprise N-[G / S]ISSS[S / G][S / A]YI[L / Y]YADSVKG-C (SEQ ID NO: 149).

[0258] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is Provided is an isolated antibody or antigen-binding portion thereof comprising heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, wherein heavy chain CDR1 does not comprise N-FTF[S / A / R][S / R][T / Y][G / S]MN-C (SEQ ID NO: 148) and / or heavy chain CDR2 does not comprise N-[G / S]ISSS[S / G][S / A]YI[L / Y]YADSVKG-C (SEQ ID NO: 149).

[0259] In some embodiments, the present disclosure specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). Provided is an isolated antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof comprises heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3, and wherein heavy chain CDR1 does not comprise N-FTF[S / A / R][S / R][T / Y][G / S]MN-C (SEQ ID NO: 148) and / or heavy chain CDR2 does not comprise N-[G / S]ISSS[S / G][S / A]YI[L / Y]YADSVKG-C (SEQ ID NO: 149).

[0260] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-ISSSXXYI-C (SEQ ID NO: 147), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 121, and a light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129, respectively; or (ii) a heavy chain CDR1 consisting of N-FTFXXXXMN-C (SEQ ID NO: 150), a heavy chain CDR2 consisting of N-XISSSXXYIXYADSVKG-C (SEQ ID NO: 151), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 124, and light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 130, 131, and 132, respectively; The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0261] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-ISSSXXYI-C (SEQ ID NO: 147), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 121, and a light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129; or (ii) a heavy chain CDR1 consisting of N-FTFXXXXMN-C (SEQ ID NO: 150), a heavy chain CDR2 consisting of N-XISSSXXYIXYADSVKG-C (SEQ ID NO: 151), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 124, and a light chain CDR1, CDR2, and CDR3 sequence set forth in SEQ ID NOs: 130, 131, and 132, respectively. The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0262] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof is (i) a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-ISSSXXYI-C (SEQ ID NO: 147), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 121, and a light chain CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 127, 128, and 129; or (ii) a heavy chain CDR1 consisting of N-FTFXXXXMN-C (SEQ ID NO: 150), a heavy chain CDR2 consisting of N-XISSSXXYIXYADSVKG-C (SEQ ID NO: 151), and a heavy chain CDR3 sequence set forth in SEQ ID NO: 124, and a light chain CDR1, CDR2, and CDR3 sequence set forth in SEQ ID NOs: 130, 131, and 132, respectively. The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0263] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-IXXXXXXX-C (SEQ ID NO: 152), and a heavy chain CDR3 consisting of N-AR[X] n=6-15 a heavy chain CDR3 sequence consisting of DX-C (SEQ ID NO: 153); and N-QS[X] n=1-3 SS[X] n=0-4 A light chain CDR1 consisting of YC (SEQ ID NO: 154), a light chain CDR2 consisting of N-XXS-C (SEQ ID NO: 155), and a light chain CDR3 consisting of N-QQXXXXP[X] n=0-1 Light chain CDR3 sequence consisting of TC (SEQ ID NO: 156) The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0264] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-IXXXXXXX-C (SEQ ID NO: 152), and a heavy chain CDR3 consisting of N-AR[X]XXXXXX-C (SEQ ID NO: 153), respectively. n=6-15a heavy chain CDR3 sequence consisting of DX-C (SEQ ID NO: 153); and N-QS[X] n=1-3 SS[X] n=0-4 A light chain CDR1 consisting of YC (SEQ ID NO: 154), a light chain CDR2 consisting of N-XXS-C (SEQ ID NO: 155), and a light chain CDR3 consisting of N-QQXXXXP[X] n=0-1 Light chain CDR3 sequence consisting of TC (SEQ ID NO: 156) The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0265] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising or consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 consisting of N-GFTFXXXX-C (SEQ ID NO: 145), a heavy chain CDR2 consisting of N-IXXXXXXX-C (SEQ ID NO: 152), and N-AR[X] n=6-15 a heavy chain CDR3 sequence consisting of DX-C (SEQ ID NO: 153); and N-QS[X] n=1-3 SS[X] n=0-4 A light chain CDR1 consisting of YC (SEQ ID NO: 154), a light chain CDR2 consisting of N-XXS-C (SEQ ID NO: 155), and a light chain CDR3 consisting of N-QQXXXXP[X] n=0-1 Light chain CDR3 consisting of TC (SEQ ID NO: 156) The present invention provides an isolated antibody or antigen-binding portion thereof, which is free of

[0266] In some embodiments, the present disclosure provides compounds that antagonize IL-27 and are selected from the group consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). or an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising a plurality of amino acids, wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 15, 37, 59, 81, 103, and 125, and the light chain variable region does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 45, 67, 89, 111, and 133.

[0267] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that antagonizes IL-27 and specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region and a light chain variable region, (i) SEQ ID NOs: 15 and 65, respectively; (ii) SEQ ID NOs: 37 and 45, respectively; (iii) SEQ ID NOs: 59 and 67, respectively; (iv) SEQ ID NOs: 81 and 89, respectively; (v) SEQ ID NOs: 103 and 111, respectively; and (vi) SEQ ID NOs: 125 and 133, respectively The present invention provides an isolated antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof is not an amino acid sequence selected from the group consisting of:

[0268] In some embodiments, the present disclosure provides antibodies that antagonize IL-27 and specifically identify an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28).

[0010] Provided is an isolated antibody or antigen-binding portion thereof that heterologously binds to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 15, 37, 59, 81, 103, and 125, wherein the heavy chain variable region does not comprise an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 15, 37, 59, 81, 103, and 125, and the light chain variable region does not comprise an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 45, 67, 89, 111, and 133.

[0269] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that antagonizes IL-27 and specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region and a light chain variable region, (i) SEQ ID NOs: 15 and 65, respectively; (ii) SEQ ID NOs: 37 and 45, respectively; (iii) SEQ ID NOs: 59 and 67, respectively; (iv) SEQ ID NOs: 81 and 89, respectively; (v) SEQ ID NOs: 103 and 111, respectively; and (vi) SEQ ID NOs: 125 and 133, respectively The present invention provides an isolated antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion is at least 90% different from an amino acid sequence selected from the group consisting of:

[0270] In some embodiments, the present disclosure provides compounds that antagonize IL-27 and are selected from the group consisting of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and G of SEQ ID NO: 2 (IL-27p28). Provided is an isolated antibody or antigen-binding portion thereof that specifically binds to an epitope comprising one or more amino acids of lu164, wherein the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, wherein the heavy chain does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 25, 47, 69, 91, 113, and 135, and the light chain does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 20, 42, 71, 93, and 1115.

[0271] In some embodiments, the present invention provides compounds that antagonize IL-27 and bind to one or more of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). and the light chain does not comprise an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 20, 42, 71, 93, and 115.

[0272] In some embodiments, the present disclosure provides compounds that antagonize IL-27 and target Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu1 of SEQ ID NO: 2 (IL-27p28). 64, wherein the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, and the heavy chain does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 29, 51, 73, 95, 117, and 139, and the light chain does not comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 71, 49, 71, 93, 115, and 137.

[0273] In some embodiments, the present disclosure provides compounds that antagonize IL-27 and are selected from the group consisting of one or more amino acids Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28). and the light chain does not comprise an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 71, 49, 71, 93, 115, and 137.

[0274] In some embodiments, the present invention provides an isolated antibody or antigen-binding portion thereof that antagonizes IL-27 and specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, and the heavy chain and the light chain are (i) SEQ ID NOs: 25 and 27, respectively; (ii) SEQ ID NOs: 47 and 49, respectively; (iii) SEQ ID NOs: 69 and 71, respectively; (iv) SEQ ID NOs: 91 and 93, respectively; (v) SEQ ID NOs: 113 and 115, respectively; and (vi) SEQ ID NOs: 135 and 137, respectively The present invention provides an isolated antibody or antigen-binding portion thereof, which does not comprise an amino acid sequence selected from the group consisting of:

[0275] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that antagonizes IL-27 and specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, and the heavy chain and the light chain are (i) SEQ ID NOs: 25 and 27, respectively; (ii) SEQ ID NOs: 47 and 49, respectively; (iii) SEQ ID NOs: 69 and 71, respectively; (iv) SEQ ID NOs: 91 and 93, respectively; (v) SEQ ID NOs: 113 and 115, respectively; and (vi) SEQ ID NOs: 135 and 137, respectively The present invention provides an isolated antibody or antigen-binding portion thereof that does not contain an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of:

[0276] In some embodiments, the disclosure provides an isolated antibody or antigen-binding portion thereof that antagonizes IL-27 and specifically binds to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28), wherein the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, and the heavy chain and the light chain are (i) SEQ ID NOs: 29 and 27, respectively; (ii) SEQ ID NOs: 51 and 49, respectively; (iii) SEQ ID NOs: 73 and 72, respectively; (iv) SEQ ID NOs: 95 and 93, respectively; (v) SEQ ID NOs: 117 and 115, respectively; and (vi) SEQ ID NOs: 139 and 137, respectively The present invention provides an isolated antibody or antigen-binding portion thereof, which does not comprise an amino acid sequence selected from the group consisting of:

[0277] In some embodiments, the present disclosure provides antibodies that antagonize IL-27 and specifically bind to an epitope comprising one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL27-p28). (iv) SEQ ID NOs: 95 and 93, respectively; (v) SEQ ID NOs: 117 and 115, respectively; and (vi) SEQ ID NOs: 139 and 137, respectively.

[0278] D. Combination Therapy In some embodiments, an IL-27 inhibitor provided by the present disclosure, e.g., an anti-IL-27 antibody or antigen-binding portion thereof, can be combined with one or more additional therapeutic agents or treatments, e.g., another therapeutic agent or treatment for cancer. For example, an IL-27 inhibitor, e.g., an anti-IL-27 antibody or antigen-binding portion thereof, can be administered to a subject (e.g., a human patient) in combination with one or more additional therapeutic agents, the combination providing a therapeutic benefit to the subject suffering from or at risk of developing cancer.

[0279] In some embodiments, the IL-27 inhibitor, e.g., an anti-IL-27 antibody, or antigen-binding portion thereof, and the one or more additional therapeutic agents are administered at the same time (e.g., simultaneously). In other embodiments, the IL-27 inhibitor, e.g., an anti-IL-27 antibody, or antigen-binding portion thereof, is administered first, and the one or more additional therapeutic agents are administered second (e.g., sequentially). In some embodiments, the one or more additional therapeutic agents are administered first, and the IL-27 inhibitor, e.g., an anti-IL-27 antibody, is administered second.

[0280] The IL-27 inhibitors described herein, e.g., anti-IL-27 antibodies or antigen-binding fragments thereof, can replace or augment previously or currently administered therapies. For example, when treating with an anti-IL-27 antibody or antigen-binding fragment thereof, the administration of one or more additional therapeutic agents can be discontinued or reduced, e.g., administered at a lower level. In some embodiments, the administration of the previous treatment can be maintained. In some embodiments, the previous treatment is maintained until the level of the anti-IL-27 antibody reaches a level sufficient to provide a therapeutic effect.

[0281] In some embodiments, the present disclosure provides methods of treating cancer in a subject, comprising administering to the subject an effective amount of an isolated antibody or antigen-binding portion thereof that specifically binds and antagonizes IL-27 provided by the present disclosure in combination with one or more additional therapeutic agents or treatments, wherein the second therapeutic agent or treatment is selected from the group consisting of chemotherapy, targeted anti-cancer therapy, oncolytic agent, cytotoxic agent, immune-based therapy, cytokine, surgery, radiation treatment, activator of costimulatory molecules, inhibitor of inhibitory molecules, vaccine, or cellular immunotherapy, biologic agent, or combination thereof.

[0282] In some embodiments, the one or more additional therapeutic agents are a PD-1 antagonist, a TIM-3 inhibitor, a LAG-3 inhibitor, a TIGIT inhibitor, a CD112R inhibitor, a TAM inhibitor, a STING agonist, a 4-1BB agonist, or a combination thereof. In some embodiments, the one or more additional therapeutic agents are a CD39 antagonist, a CD73 antagonist, a CCR8 antagonist, or a combination thereof. In some embodiments, the anti-CD73 is any anti-CD73 antibody disclosed in, for example, U.S. Patent Application Publication No. 2019 / 0031766 A1, the specification of which is incorporated herein by reference in its entirety. In some embodiments, the anti-CD39 is any anti-CD39 antibody disclosed in, for example, International Publication No. WO2019 / 178269 A2, the specification of which is incorporated herein by reference in its entirety.

[0283] In some embodiments, the one or more additional therapeutic agents are PD-1 antagonists. In some embodiments, the PD-1 antagonist is selected from the group consisting of PDR001, nivolumab, pembrolizumab, pidilizumab, tislelizumab, zimblerimab, MEDI0680, REGN2810, TSR-042, PF-06801591, and AMP-224. In certain embodiments, the one or more additional therapeutic agents are PD-L1 inhibitors. In some embodiments, the PD-L1 inhibitor is selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559. In some embodiments, the present disclosure provides methods of enhancing one or more activities of an anti-PD-1 antibody (e.g., enhancing PD-1-mediated cytokine secretion; enhancing anti-PD-1-mediated TNFα secretion; enhancing anti-PD-1-mediated IL-6 secretion from cells exposed to the anti-PD-1 antibody), comprising exposing cells to an antibody, or antigen-binding portion thereof, provided by the present disclosure, simultaneously or sequentially with the anti-PD-1 antibody, thereby enhancing one or more activities of the anti-PD-1 antibody.

[0284] In some embodiments, the one or more additional therapeutic agents are selected from the group consisting of sunitinib (SUTENT®), cabozantinib (CABOMETYX®), axitinib (INLYTA®), lenvatinib (LENVIMA®), everolimus (AFINITOR®), bevacizumab (AVASTIN®), epacadostat, NKTR-214 (CD-122 biased agonist), tivozanib (FOTIVDA®), abexinostat, ipilimumab, rivaroxaban ... ramucirumab (YERVOY®), tremelimumab, pazopanib (VOTRIENT®), sorafenib (NEXAVAR®), temsirolimus (TORISEL®), ramucirumab (CYRAMZA®), niraparib, savolitinib, borolinib (X-82), regorafenib (STIVARGO®), donafenib (multikinase inhibitor), camrelizumab (SHR-1210), pexastimogene devasilepvec (pexastimogene devacirepvec (JX-594), ramucirumab (CYRAMZA®), apatinib (YN968D1), encapsulated doxorubicin (THERMODOX®), tivantinib (ARQ197), ADI-PEG 20, binimetinib, apatinib mesylate, nintedanib, lirilumab, nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFIMZI®), cemiplimab-rwlc (LIBTAYO®), tislelizumab, and / or spartalizumab.

[0285] In some embodiments, the one or more additional therapeutic agents is a TIM-3 inhibitor, optionally wherein the TIM-3 inhibitor is MGB453 or TSR-022.

[0286] In some embodiments, the one or more additional therapeutic agents is a LAG-3 inhibitor, optionally wherein the LAG-3 inhibitor is selected from the group consisting of LAG525, BMS-986016, and TSR-033.

[0287] In some embodiments, the one or more additional therapeutic agents are TIGIT inhibitors. In some embodiments, the one or more additional therapeutic agents are CD112R inhibitors. In some embodiments, the one or more additional therapeutic agents are TAM (Axl, Mer, Tyro) inhibitors. In some embodiments, the one or more additional therapeutic agents are STING agonists. In some embodiments, the one or more additional therapeutic agents are 4-1BB agonists.

[0288] In some embodiments, the one or more additional therapeutic agents are a tyrosine kinase inhibitor, an agent that targets the adenosine axis (e.g., a CD39 antagonist, a CD73 antagonist, or an A2AR, A2BR, or dual A2AR / A2BR antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof.

[0289] 1. Combination with chemotherapy In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds IL-27, and a chemotherapeutic agent. Chemotherapeutic agents suitable for combination and / or co-administration with the compositions of the present disclosure include, for example, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyantransinedione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or homologs thereof. Additional agents include, for example, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioTEPA, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlordiamineplatinum(II) (DDP), procarbazine, These include cyclosporine, altretamine, cisplatin, carboplatin, oxaliplatin, nedaplatin, satraplatin, or triplatin tetranitrate), anthracyclines [e.g., daunorubicin (formerly known as daunomycin) and doxorubicin], antibiotics [e.g., dactinomycin (formerly known as actinomycin), bleomycin, mithramycin, and anthramycin (AMC)], antimitotic agents (such as vincristine and vinblastine), and temozolomide.

[0290] 2. Combination with PD-1 / PD-L1 antagonist In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and one or more PD-1 antagonists. In some embodiments, the one or more PD-1 antagonists specifically bind to human PD-1 or PD-L1 and inhibit the biological activity and / or downstream pathway(s) of PD-1 / PD-L1 and / or cellular processes mediated by human PD-1 / PD-L1 signaling, or other human PD-1 / PD-L1-mediated functions.

[0291] Accordingly, provided herein are PD-1 antagonists that directly or allosterically block, antagonize, suppress, inhibit, or reduce the biological activity of PD-1 / PD-L1, including downstream pathways and / or cellular processes mediated by PD-1 / PD-L1 signaling, such as receptor binding and / or eliciting a cellular response to PD-1 / PD-L1. Also provided herein are PD-1 antagonists that reduce the amount or quantity of human PD-1 or PD-L1 produced by a cell or a subject.

[0292] In some embodiments, the present disclosure provides PD-1 antagonists that bind to human PD-1 and prevent, inhibit, or reduce PD-L1 binding to PD-1. In some embodiments, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and prevents translation. In some embodiments, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and causes degradation and / or turnover.

[0293] In some embodiments, the PD-1 antagonist inhibits PD-1 signaling or function. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L2. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist specifically binds to PD-1. In some embodiments, the PD-1 antagonist specifically binds to PD-L1. In some embodiments, the PD-1 antagonist specifically binds to PD-L2.

[0294] In some embodiments, the PD-1 antagonist inhibits the binding of PD-1 to its cognate ligand. In some embodiments, the PD-1 antagonist inhibits the binding of PD-1 to PD-L1, the binding of PD-1 to PD-L2, or the binding of PD-1 to both PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist does not inhibit the binding of PD-1 to its cognate ligand.

[0295] In some embodiments, the PD-1 antagonist is an isolated antibody (mAb) or antigen-binding fragment thereof that specifically binds to PD-1 or PD-L1. In some embodiments, the PD-1 antagonist is an antibody or antigen-binding fragment thereof that specifically binds to human PD-1. In some embodiments, the PD-1 antagonist is an antibody or antigen-binding fragment thereof that specifically binds to human PD-L1. In some embodiments, the PD-1 antagonist is an antibody or antigen-binding fragment thereof that binds to human PD-L1 and inhibits binding of PD-L1 to PD-1. In some embodiments, the PD-1 antagonist is an antibody or antigen-binding fragment thereof that binds to human PD-1 and inhibits binding of PD-L1 to PD-1.

[0296] Several immune checkpoint antagonists that inhibit or disrupt the interaction of PD-1 with one or both of its ligands, PD-L1 and PD-L2, are in clinical development or are currently available to clinicians for the treatment of cancer.

[0297] Examples of anti-human PD-1 antibodies or antigen-binding fragments thereof that may comprise PD-1 antagonists in any of the compositions, methods, and uses provided by the present disclosure include, but are not limited to, KEYTRUDA® (pembrolizumab, MK-3475, h409A11; see U.S. Patent Nos. 8,952,136, 8,354,509, 8,900,587, and EP 2,170,959, all of which are incorporated herein by reference in their entireties). Merck), OPDIVO® (nivolumab, BMS-936558, MDX-1106, ONO-4538; see U.S. Patent Nos. 7,595,048, 8,728,474, 9,073,994, 9,067,999, EP 1,537,878, 8,008,449, 8,779,105, and EP 2,161,336, all of which are incorporated by reference in their entireties; Bristol Myers Squibb), MEDI0680 (AMP-514), BGB-A317 and BGB-108 (BeiGene), 244C8 and 388D4 (see WO2016106159, which is incorporated herein by reference in its entirety; Enumeral Biomedical), PDR001 (Novartis), and REGN2810 (Regeneron). Accordingly, in some embodiments, the PD-1 antagonist is pembrolizumab. In some embodiments, the PD-1 antagonist is nivolumab. In some embodiments, the methods disclosed herein comprise administering an antibody, or antigen-binding portion thereof, that specifically binds IL-27 and pembrolizumab. In some embodiments, the methods disclosed herein comprise administering an antibody, or antigen-binding portion thereof, that specifically binds IL-27 and nivolumab.

[0298] Examples of anti-human PD-L1 antibodies or antigen-binding fragments thereof that may comprise PD-1 antagonists in any of the compositions, methods, and uses provided by the present disclosure include, but are not limited to, BAVENCIO® (avelumab, MSB0010718C, see WO2013 / 79174, which is incorporated herein by reference in its entirety; Merck / Pfizer), IMFINZI® (durvalumab, MEDI4736), TECEN® (anti-PD-L1 antibodies, ... Examples of PD-1 antagonists include TRIQ® (atezolizumab, MPDL3280A, RG7446; see WO2010 / 077634, which is incorporated herein by reference in its entirety; Roche), MDX-1105 (BMS-936559, 12A4; see U.S. Patent No. 7,943,743 and WO2013 / 173223, both of which are incorporated herein by reference in their entireties; Medarex / BMS), and FAZ053 (Novartis). Thus, in some embodiments, the PD-1 antagonist is avelumab. In some embodiments, the PD-1 antagonist is durvalumab. In some embodiments, the PD-1 antagonist is atezolizumab.

[0299] In some embodiments, the PD-1 antagonist is an immunoadhesin that specifically binds to human PD-1 or human PD-L1, e.g., a fusion protein comprising the extracellular portion or PD-1-binding portion of PD-L1 or PD-L2 fused to a constant region, such as the Fc region, of an immunoglobulin molecule. Examples of immunoadhesins that specifically bind to PD-1 are described in WO2010 / 027827 and WO2011 / 066342, both of which are incorporated by reference in their entireties. In some embodiments, the PD-1 antagonist is AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein that specifically binds to human PD-1.

[0300] One of skill in the art will appreciate that any PD-1 antagonist that binds to PD-1 or PD-L1 and disrupts the PD-1 / PD-L1 signaling pathway is suitable for the compositions, methods, and uses disclosed herein.

[0301] In some embodiments, the PD-1 / PD-L1 antagonist is a small molecule, nucleic acid, peptide, peptidomimetic, protein, carbohydrate, carbohydrate derivative, or glycopolymer. Exemplary small molecule PD-1 inhibitors are described in Zhan et al., (2016) Drug Discov Today 21(6):1027-1036.

[0302] 3. Combination with TIM-3 inhibitors In some embodiments, the methods disclosed herein include administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a TIM-3 inhibitor. The TIM-3 inhibitor can be an antibody, antigen-binding fragment thereof, immunoadhesin, fusion protein, or oligopeptide. In some embodiments, the TIM-3 inhibitor is selected from MGB453 (Novartis), TSR-022 (Tesaro), or LY3321367 (Eli Lilly). In some embodiments, the anti-IL-27 antibody or antigen-binding portion thereof is administered in combination with MGB453. In some embodiments, the anti-IL-27 antibody or antigen-binding portion thereof is administered in combination with TSR-022.

[0303] 4. Combination with LAG-3 inhibitors In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and an LAG-3 inhibitor. In some embodiments, the LAG-3 inhibitor is an antibody, antigen-binding fragment thereof, immunoadhesin, fusion protein, oligopeptide, or any combination thereof. In some embodiments, the LAG-3 inhibitor is selected from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), TSR-033 (Tesaro), MK-4280 (Merck & Co.), or REGN3767 (Regeneron).

[0304] 5. Other combinations In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a TIGIT inhibitor. In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a kinase inhibitor (e.g., a tyrosine kinase inhibitor (TKI)). In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a CD112R inhibitor. In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a TAM receptor inhibitor. In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and a STING agonist and / or a 4-1BB agonist. In some embodiments, an anti-IL-27 antibody, or antigen-binding portion thereof, provided by the present disclosure is combined (e.g., administered in combination) with a tyrosine kinase inhibitor, an agent that targets the adenosine axis (e.g., a CD39 antagonist, a CD73 antagonist, or an A2AR, A2BR, or dual A2AR / A2BR antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof.

[0305] In some embodiments, the methods disclosed herein comprise administering an IL-27 inhibitor, e.g., an antibody or antigen-binding portion thereof that specifically binds to IL-27, and cell therapy. In some embodiments, the cell therapy comprises engineered immune cell therapy. In some embodiments, the cell therapy comprises chimeric antigen receptor (CAR)-engineered immune cell therapy, e.g., CART therapy. In some embodiments, the cell therapy comprises engineered T cell receptor (TCR) immune cell therapy. In some embodiments, the cell therapy comprises allogeneic tumor-infiltrating lymphocyte (TIL) therapy. [Example]

[0306] While the present disclosure has been described with reference to specific embodiments thereof, it will be recognized by those skilled in the art that various changes may be made and equivalents substituted without departing from the true spirit and scope of the present disclosure. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present disclosure. All such modifications are intended to be within the scope of the present disclosure. [Example 1]

[0307] Detection of WSX-1 in tumor samples Unstained formalin-fixed, paraffin-embedded (FFPE) slides were dewaxed and pretreated in Bond Epitope Retrieval 2 solution (Leica Biosystems, Buffalo Grove, IL) for 20 minutes in a Leica Bond RX autostainer (Leica Biosystems) according to the manufacturer's recommended protocol. DAKO protein block (Agilent Technologies, Santa Clara, CA) was applied to the slides and incubated for 15 minutes. Monoclonal rabbit anti-human WSX-1 primary antibody (catalog no. ab281998, Abcam, Cambridge, MA) was diluted 1 / 50 in Bond antibody diluent (catalog no. AR9352, Leica Biosystems) and dispensed onto slides. The slides were incubated at room temperature for 30 minutes, followed by a ready-to-use secondary antibody (HRP-conjugated goat anti-rabbit IgG polymer, Bond Refine Detection Kit, catalog no. DS9800, Leica Biosystems) for 8 minutes at room temperature, followed by a 5-minute peroxidase block (Bond Polymer Refine Detection Kit, catalog no. DS9800, Leica Biosystems). Signals were detected by a 10-minute DAB incubation followed by a 10-minute DAB enhancer incubation (Bond Polymer Refine Detection Kit, catalog no. DS9800, Leica Biosystems). Slides were counterstained with hematoxylin for 10 minutes. Slides were manually dehydrated and coverslipped using Polymount Xylene Mounting Medium (Cat. No. 24176, Polysciences, Warrington, PA).

[0308] WSX-1 mRNA expression was detected in various cancer cell lines (Figure 1A). IHC analysis of lung adenocarcinoma, lung SCC, ovarian cancer, head and neck squamous cell carcinoma, and TNBC revealed positive WSX-1 expression in tumor cells in multiple indications, which was maintained at lymph node metastatic sites (Figures 1B-1J). [Example 2]

[0309] Detection of IL-27 in tumor samples Unstained FFPE slides were dewaxed and pretreated with Bond Epitope Retrieval 2 solution (Leica Biosystems, Buffalo Grove, IL) for 10 minutes in a Leica Bond RX autostainer (Leica Biosystems) according to the manufacturer's recommended protocol. DAKO protein block (Agilent Technologies, Santa Clara, CA) was applied to the slides and incubated for 15 minutes. Polyclonal goat anti-human IL-27 primary antibody (catalog no. AF2526, R&D Systems, Minneapolis, MN) was diluted to 3.33 μg / ml (1:60 dilution from a 200 μg / ml stock) in Bond Antibody Diluent (catalog no. AR9352, Leica Biosystems) and dispensed onto slides. The slides were incubated at room temperature for 60 minutes, followed by a ready-to-use secondary antibody (ImmPRESS-HRP Horse Anti-Goat IgG Polymer, catalog no. MP-7405, Vector Laboratories, Inc., Burlington, CA) for 8 minutes at room temperature, followed by a 5-minute peroxidase block (Bond Polymer Refine Detection Kit, catalog no. DS9800, Leica Biosystems). Signal was detected by a 10-minute DAB incubation and a 10-minute DAB enhancer incubation (Bond Polymer Refine Detection Kit, catalog no. DS9800, Leica Biosystems). Slides were counterstained with hematoxylin for 5 minutes. A detailed Bond RX staining protocol is provided (Protocol Name: Goat60_AZ2). Slides were manually dehydrated and coverslipped using Polymount Xylene Mounting Medium (Cat. No. 24176, Polysciences, Warrington, PA).

[0310] Immunohistochemistry (IHC) for IL-27 was performed on formalin-fixed, paraffin-embedded (FFPE) tumor samples in tissue microarray (TMA) format. Staining reveals positive cells within the tumor microenvironment (TME) morphologically consistent with tumor-associated macrophages (TAMs) across various cancer types (Figure 2A-2F). Quantitative image analysis of digitally scanned slides was used to quantify the density of IL-27+ cells across multiple solid tumor types (Figure 2G). An sc-RNA-seq analysis of the Immune Atlas on various cancer tumor cells for global IL-27 expression across all cancer types studied is shown in Figure 2H.

[0311] IHC for PD-L1 and IL-27 was performed on serial sections of FFPE tumor samples in TMA format. PD-L1 staining was semiquantitatively scored using the composite percentage score (CPS) system, and the density of IL-27+ cells was quantified using image analysis of digitally scanned slides. The density of IL-27+ cells positively correlated with PD-L1 expression in NSCLC (Figure 3A), gastric cancer (Figure 3B), and HCC (Figure 3C). Examples of PD-L1 and IL-27 IHC in gastric cancer samples with various CPS scores are shown in Figures 3D–3I. While the CPS score correlated with the density of IL-27+ cells (Figures 3D, 3E, 3G, and 3H), a proportion of tumor samples with a CPS of less than 1 (PD-L1 negative) contained IL-27+ cells (Figures 3F and 3I).

[0312] IHC for IL-27, WSX-1, and PD-L1 was performed on serial sections of normal human tonsil and lung SCC, and positive staining was observed for all stains (Figures 4A-4I). [Example 3]

[0313] In vivo analysis of patient response to anti-IL-27 antibody monotherapy To identify correlations between IL-27 expression and / or WSX-1 expression, tumor samples are collected from patients before administering anti-IL-27 antibody therapy. The tumor samples are assayed for IL-27 and / or WSX-1 expression according to the methods disclosed herein, for example, the methods disclosed in Examples 1 and 2. The patients are administered anti-IL-27 antibodies according to the methods disclosed herein. Retrospective analyses are performed to identify correlations between IL-27 expression and / or WSX-1 expression and patient overall survival, objective response rate, progression-free survival, and tumor size. [Example 4]

[0314] In vivo analysis of patient response to anti-IL-27 antibody combination therapy To identify correlations between IL-27 expression and / or WSX-1 expression, tumor samples are collected from patients before administering anti-IL-27 antibody therapy. The tumor samples are assayed for IL-27 and / or WSX-1 expression according to the methods disclosed herein, e.g., the methods disclosed in Examples 1 and 2. The patients are administered (i) an anti-IL-27 antibody and (ii) an additional anti-cancer agent disclosed herein (e.g., an immune checkpoint inhibitor, e.g., an anti-PD-1 antibody, chemotherapy, radiation therapy, cell-based immunotherapy, or any combination thereof) according to the methods disclosed herein. Retrospective analysis is performed to identify correlations between IL-27 expression and / or WSX-1 expression and patient overall survival, objective response rate, progression-free survival, and tumor size. [Example 5]

[0315] Detection of IL-27 and WSX-1 in NSCLC and HCC samples IHC for IL-27 was performed on FFPE NSCLC tumor samples obtained as whole sections from individual lobectomy specimens. IL-27 IHC shows aggregates of IL-27+ macrophages that are heterogeneously distributed throughout the tumor mass (Figure 5). Center image: 0.9x magnification; inset: 12x magnification.

[0316] IHC for WSX-1 was performed on FFPE NSCLC tumor samples obtained as whole sections from individual lobectomy specimens. WSX-1 IHC demonstrates WSX-1 immunocytostaining in the peritumoral immune cell infiltrate (Figure 6A, 0.55x magnification). WSX-1 immunocytostaining is prominent in tertiary lymphoid structures (TLS) (Figure 6B, 3.5x magnification). Follicular dendritic cells (FDCs) within the germinal centers of TLS are positive for WSX-1 (Figure 6C, 12x magnification). WSX-1 immunocytostaining is also seen outside the TLS (Figure 6D, 20x magnification).

[0317] IHC for IL-27 and WSX-1 was performed on FFPE NSCLC tumor samples obtained as whole sections from individual lobectomy specimens. IHC demonstrated immunocytochemical staining for IL-27 and WSX-1 within the same region of the tumor microenvironment (TME), most prominent within the TLS (Figure 7A, matched fields from the same specimen, 7x magnification). Immune cell staining for IL-27 and WSX-1 was scored semiquantitatively using the following pathology scoring system: 0, no staining or rare positive immune cells; 1+, positive immune cells in less than 33% of the tumor tissue area; 2+, positive immune cells in 33–66% of the tumor tissue area; and 3+, positive immune cells in more than 66% of the tumor tissue area (Figure 7B).

[0318] IHC for IL-27 and WSX-1 was performed on FFPE samples of NSCLC draining lymph nodes obtained from individual lobectomy specimens and showed abundant immune cell staining for both markers (Figure 8A, 8B). Specifically, abundant IL-27+ macrophages were present within the interfollicular regions, abundant WSX-1+ immune cells were present within the interfollicular regions, and prominent WSX-1+ FDCs were present within germinal centers. This was observed not only in tumor-containing draining lymph nodes (lymph node metastasis; Figure 8A, matched field from the same specimen, 10x magnification) but also in tumor-free draining lymph nodes (Figure 8B, matched field from the same specimen, 7x magnification). In contrast, IHC for IL-27 and WSX-1 on control lymph nodes showed minimal immune cell staining for IL-27 and WSX-1 (Figure 8C, 15x magnification). n = 2–5 specimens / group (5 tumor-bearing NSCLC draining lymph nodes, 2 tumor-free NSCLC draining lymph nodes, and 5 control lymph nodes).

[0319] A subset of NSCLC samples shows tumor cell staining for IL-27 or WSX-1 (Figures 9A, 9B, and 9C). In 25.0% of NSCLC cases, membranous (cytoplasmic and / or membranous) tumor cell staining for IL-27 is seen in at least 1% of tumor cells (Figure 9A, 20x magnification, Figure 9C). In 45.5% of NSCLC cases, membranous tumor cell staining for WSX-1 is seen in at least 1% of tumor cells (Figure 9B, 20x magnification, Figure 9C). Bars represent the mean of n=44 NSCLC lobectomy specimens (Figure 9C).

[0320] IHC for CD8 shows that in NSCLC cases with an immune exclusion pattern (CD8+ T cells are primarily confined to the peritumoral immune cell infiltrate with minimal infiltration into the tumor mass), there is prominent immune cell staining for IL-27 and WSX-1 in the immune cell infiltrate surrounding the tumor where CD8+ T cells accumulate (Figure 10A, magnification 2.2x). Similarly, in NSCLC cases with a more subtle immune exclusion pattern (CD8+ T cells infiltrate the main tumor mass but are essentially confined to the stroma, with no significant infiltration into tumor nests), aggregates of IL-27+ macrophages are seen in the stromal area where CD8+ T cells accumulate (Figure 10B, magnification 4x [top] and 10x [bottom]). Immune cell staining for PD-L1 is also seen in the same stromal area (Figure 10B).

[0321] In NSCLC, immune cell staining for PD-L1 is seen within the same stromal regions containing aggregates of IL-27+ macrophages (Figure 11A, lung adenocarcinoma, 4x magnification [top] and 20x magnification [bottom], and Figure 11B, lung squamous cell carcinoma, 6x magnification [top] and 18x magnification [bottom]). Within these regions, the morphology of PD-L1+ immune cells is very similar to that of IL-27+ macrophages, suggesting that macrophage populations positive for both markers may exist within the TME of NSCLC.

[0322] In NSCLC, IL-27 and WSX-1 expression generally correlated with PD-L1 expression (Figures 12A, 12B, 12C, and 12D). When PD-L1 status was classified using the tumor proportion score (TPS) system, which only considers PD-L1 expression on tumor cells, immune cell expression of IL-27 and WSX-1 tended to be higher in cases with high PD-L1 expression (Figure 12A). When PD-L1 status was classified using the combined proportion score (CPS) system, which considers PD-L1 expression on tumor cells and immune cells, immune cell expression of IL-27 and WSX-1 was statistically significantly higher in cases with high PD-L1 expression (Figure 12B). Data points are the mean ± SEM of n = 24 NSCLC lobectomy specimens. IHC scores for IL-27 and WSX-1 were scored semiquantitatively using the following pathological scoring system (described in Figure 7): 0, no staining or rare positive immune cells; 1+, positive immune cells in less than 33% of the tumor tissue area; 2+, positive immune cells in 33-66% of the tumor tissue area; and 3+, positive immune cells in more than 66% of the tumor tissue area. The scores were calculated by Student's T-test. * :p<0.01, ** p<0.01. All other comparisons were not statistically significant. Similarly, tumor cell staining for IL-27 and WSX-1 was frequently observed in cases with high PD-L1 expression, especially when PD-L1 expression was classified using the CPS system (Figures 12C and 12D). Data points are the percentage of positive cases in n=24 NSCLC lobectomy specimens. Positivity is defined as at least 1% of tumor cells exhibiting membranous (IL-27) or membranous (WSX-1) tumor cell staining.

[0323] In NSCLC, there is a general trend toward higher immune cell expression of IL-27 and WSX-1 in patients who subsequently respond to immune checkpoint blockade (ICP) (Figure 13A). There is also a general trend between response to ICP and PD-L1 status (Figure 13B), but this trend is statistically weaker than the trend between response to ICP and immune cell expression of IL-27 and WSX-1. Data points are mean ± SEM of n=24 NSCLC lobectomy specimens. IHC was performed on archival resection specimens from patients subsequently treated with ICP, and the best complete response rates were recorded. IHC staining for IL-27 and WSX-1 was semiquantitatively scored using the following pathology scoring system (as previously described in Figures 7 and 11): 0, no staining or rare positive immune cells; 1+, positive immune cells in less than 33% of the tumor tissue area; 2+, positive immune cells in 33-66% of the tumor tissue area; 3+, positive immune cells in more than 66% of the tumor tissue area. TPS: tumor percentage score; CPS: composite percentage score. *p<0.01 by Student's t-test. All other comparisons were not statistically significant. PD: progressive disease, SD: stable disease, PR: partial response, CR: complete response. The number of patients in each response category is indicated below the response label. All scores (TPS, CPS, IL-27 score, and WSX-1 score) were generated by a pathologist blinded to the response data. Patients were divided into those who received ICP as first-line therapy (Figure 13C) and those who received ICP as second-line or more advanced therapy (Figure 13D). Note that the number of patients in each cohort was small, but trends between response to ICP and immune cell expression of IL-27 and WSX-1 were observed only in patients receiving first-line therapy. Data points are the mean ± SEM of n=12 NSCLC lobectomy specimens in each cohort. All other details are the same as in Figures 13A and 13B. IHC of IL-27 on archival specimens from NSCLC patients subsequently treated with SRF388 shows that the highest number of IL-27+ macrophages was found in specimens from patients who experienced a partial response to SRF388 monotherapy (Figures 14A and 14B). This patient was the only NSCLC patient who responded to SRF388 monotherapy and from whom archival tumor samples were available for IL-27 IHC. This observation is observed when specimens are scored using the IL-27+ macrophage score by a pathologist (as previously described in Figures 7, 11, and 12) (Figure 14A) and also when specimens are scored by a contract research organization (CRO) using a more accurate scoring system (positive IL-27+ immune cells as a percentage of tumor area) (Figure 14B). The latter scoring system indicates that IL-27+ macrophages are less abundant in patients who did not respond to SRF388 monotherapy and therefore more significantly illustrates the difference in abundance of IL-27+ macrophages between archived specimens from patients who did and did not respond to SRF388 monotherapy. All scores were generated blinded to the response data.Squares indicate archival biopsy specimens; circles indicate archival resection specimens. Black circles / squares indicate partial responses to SRF388 monotherapy; gray circles / squares indicate stable disease; and white circles / squares indicate progressive disease. Note that for patient 2, two archival specimens (lung resection specimen and lymph node metastasis resection specimen) were available; the line for patient 2 represents the average of the two specimens. In patient 2, IL-27 IHC on the archival lung resection specimen revealed numerous aggregates of IL-27+ macrophages in the TME (Figure 14C, 20x magnification), including those adjacent to tertiary lymphoid structures (TLS) (Figure 14D, 15x magnification; asterisks indicate TLS). IL-27 IHC on the archival lymph node metastasis specimen from patient 2 showed abundant IL-27+ macrophages in the residual lymph node tissue (Figure 14E, 8x magnification; Figure 14F, 20x magnification).

[0324] IHC for IL-27 in archival specimens from hepatocellular carcinoma (HCC) patients subsequently treated with SRF388 in combination with atezolizumab and bevacizumab demonstrates the presence of IL-27+ macrophages in the TME in all patients who responded to the triple combination treatment (among those with archival specimens available for IL-27 IHC) (Figure 15A). IL-27+ macrophage scores were generated by a pathologist blinded to the response data using the same scoring system as previously described in Figures 7, 11, 12, and 13. Scoring using a more accurate scoring system generated by the CRO is pending. Squares indicate archival biopsy specimens; circles indicate archival resection specimens. Black circles / squares indicate partial or complete responses to SRF388 treatment in combination with atezolizumab and bevacizumab; gray circles / squares indicate stable disease; and white circles / squares indicate progressive disease. IHC for IL-27 in patients who responded to SRF388 combination treatment typically showed IL-27-positive macrophages infiltrating the tumor mass in a single-cell manner, located within sinusoidal spaces closely adjacent to tumor cells; this pattern is very similar to that observed within the sinusoidal spaces of Kupffer cells (tissue-resident macrophages of the liver) in benign liver tissue (Figure 15B, 20x magnification; Figures 15C and 15D, insets focusing on IL-27+ macrophages). However, IL-27 IHC in one case (patient 2) showed distinct aggregates of IL-27+ macrophages within the peritumoral or stromal immune cell infiltrate, which were also seen in NSCLC specimens and closely resembled the IL-27 IHC pattern described above (Figures 15E and 15F; 20x magnification). [Example 6]

[0325] Identification of IL-27-dependent biomarkers in lymphocytes, NK cells, and myeloid cells in peripheral blood and the tumor microenvironment Human PBMCs treated with IL-27, interferon (IFN), or the STING pathway agonist cGAMP were analyzed by single-cell RNA sequencing (Figure 16A). Human PBMCs collected from healthy donors were stimulated in vitro with anti-CD3 (0.25 μg / mL) in the presence or absence of recombinant human rhIL-27, rhIFNA2, rhIFNB1, or rhIFNG (all at 100 ng / mL). After 16 and 72 hours, cells were processed for scRNA-seq (10x Genomics). Seurat-based clustering was used to identify cell subsets assigned based on differential gene expression. A UMAP display of aggregated data from all conditions identifies distinct cell populations. Immunohistochemistry (IHC) was used to evaluate IL-27 and its receptors WSX-1 (IL-27RA), PD-L1, and GBP5 in human tumors (Figure 16B). As shown in Figures 17A-C, IL-27 and interferon are expressed in various immune cell types from activated PBMCs, and interferon can upregulate IL-27 expression. Subpopulations of immune cells were found to differentially express and respond to IL-27 and IFN: pDCs transiently express IFNα / IFNβ, T cells and NK cells produce IFNγ, and myeloid cells express IL-27 (Figure 17B). IL-27 and interferon also upregulate the expression of several canonical interferon-stimulated genes (Figures 18A-18C). Furthermore, IL-27 and interferon have been shown to commonly upregulate checkpoint receptor expression in various cell types (Figure 19A), but also have unique properties that alter cytokine expression (Figure 19B).

[0326] Additional genes commonly upregulated by IL-27 and interferons show high expression in a variety of cell types after stimulation with individual cytokines (FIGS. 20A and 20B).

[0327] STAT1 phosphorylation was then measured by flow cytometry in human PBMCs after 30 min of incubation with the indicated cytokines IL-27 and type 1 IFNs. STAT1 phosphorylation was significantly induced in lymphocytes and NK cells, but not in type 2 IFNs. IFNα / IFNβ signaling via STAT1 and STAT3 phosphorylation was evident in all immune cells, whereas IL-27 signaling was more restricted to T cells and NK cells, and IFNγ-induced signaling was primarily observed in myeloid cells (Figures 21A-21C).

[0328] Many canonical IFN-responsive genes were induced by both IFN and IL-27, but biased gene expression signatures were enriched across various cell types (Figures 22A-22C). For example, IL-27 stimulation resulted in differential expression of GBP5 and IRF1 in T cells and NK cells, IFNβ resulted in expression of IFIT1 and MX2 in T cells, NK cells, and monocytes, while IFNγ resulted in upregulation of SOCS1 and CXCL9 in monocytes (Figure 22C). IL-27 and IFNα / IFNβ share similar abilities to inhibit proinflammatory cytokine secretion and increase PD-L1 expression on T cells, but these functions are not evident upon IFNγ stimulation. Interestingly, while GBP5 and IRF1 are primarily upregulated by IL-27 in T / NK cells, these genes show enhancement by IFNγ in myeloid cells.

[0329] We analyzed GBP5 transcript expression in T cells and NK cells after stimulation with IL-27 or type 1 or type 2 IFN (Figure 23A). Furthermore, flow cytometry analysis of intracellular GBP5 expression in various cell types was performed after 24 hours of stimulation of PBMCs with the indicated cytokines, demonstrating differential expression when sorted by FAC among CD4+, CD8+, or NK cells (Figure 23B). These data suggest that IL-27 upregulates GBP5 expression in T cells and NK cells ex vivo.

[0330] Finally, IHC in NSCLC patient samples showed that IL-27+ macrophages colocalized with PD-L1+ immune cells in GBP5+ T cell-enriched areas within the TME, suggesting IL-27-dependent signaling in the NSCLC TME (Figure 23C).

[0331] These studies highlight the complexity, redundancy, and unique properties of interferon and IL-27 signaling across various immune cells. In contrast to IFNγ, IL-27 shares signaling capabilities with type 1 interferons in T and NK cells.

Claims

1. A method for treating a tumor in a subject, comprising administering to the subject an IL-27 inhibitor, wherein the tumor is identified as a WSX-1 positive tumor.

2. 1. A method for treating a tumor in a subject in need thereof, comprising: (i) identifying subjects with WSX-1-positive tumors; and (ii) administering to the subject an IL-27 inhibitor. A method comprising:

3. 3. The method of claim 1 or 2, wherein the WSX-1 positive tumor is identified by detecting WSX-1 expression in a tumor sample obtained from the subject.

4. A method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, the method comprising detecting WSX-1 expression in a tumor sample obtained from the subject.

5. 5. The method of claim 4, further comprising administering an IL-27 inhibitor to the subject identified as having a WSX-1 positive tumor.

6. The method according to any one of claims 3 to 5, wherein the tumor sample obtained from the subject is a tumor tissue biopsy.

7. The method according to any one of claims 3 to 6, wherein the tumor sample obtained from the subject is a formalin-fixed, paraffin-embedded tumor sample.

8. The method of any one of claims 3 to 7, wherein the tumor sample obtained from the subject contains tumor cells, tumor-infiltrating immune cells, or both.

9. 9. The method of any one of claims 3 to 8, wherein at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of the cells in the tumor sample express WSX-1.

10. The method of any one of claims 3 to 9, wherein at least about 1% of cells in the tumor sample express WSX-1.

11. The method of any one of claims 3 to 10, wherein WSX-1 expression is detected using an immunohistochemistry (IHC) assay.

12. The method of any one of claims 3 to 11, wherein WSX-1 expression is detected using an automated IHC assay.

13. The method of any one of claims 3 to 12, wherein WSX-1 expression is scored using the Tumor Proportion Score (TPS) and / or the Combined Positive Score (CPS).

14. 14. The method of claim 13, wherein the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50%, or at least 60%.

15. The method of any one of claims 3 to 14, wherein WSX-1 expression is detected by contacting the tumor sample with an antibody or antigen-binding portion thereof that specifically binds to human WSX-1.

16. A method for treating a tumor in a subject, comprising administering to the subject an IL-27 inhibitor, wherein one or more immune cells in a tumor sample obtained from the subject express IL-27.

17. 1. A method for treating a tumor in a subject in need thereof, comprising: (i) identifying a subject having a tumor in which one or more immune cells in a tumor sample obtained from the subject express IL-27; and (ii) administering to the subject an IL-27 inhibitor. A method comprising:

18. A method for identifying a human subject suffering from a tumor suitable for treatment with an IL-27 inhibitor, the method comprising detecting IL-27 expression in a tumor sample obtained from the subject.

19. 20. The method of claim 18, further comprising administering an IL-27 inhibitor to the subject identified as having a tumor sample containing one or more immune cells that express IL-27.

20. The method of any one of claims 16 to 19, wherein the tumor sample obtained from the subject is a tumor tissue biopsy.

21. The method of any one of claims 16 to 20, wherein the tumor sample obtained from the subject is a formalin-fixed, paraffin-embedded tumor sample.

22. The method of any one of claims 16 to 21, wherein the tumor sample obtained from the subject comprises tumor cells, tumor-infiltrating immune cells, or both.

23. The method of any one of claims 16 to 22, wherein the one or more immune cells in the tumor sample comprise macrophages.

24. 24. The method of any one of claims 16-23, wherein at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% of immune cells in the tumor sample express IL-27.

25. The method of any one of claims 16 to 24, wherein at least about 1% of immune cells in the tumor sample express IL-27.

26. The method of any one of claims 18 to 25, wherein IL-27 expression is detected using an immunohistochemistry (IHC) assay.

27. The method of any one of claims 18 to 26, wherein IL-27 expression is detected using an automated IHC assay.

28. The method of any one of claims 18 to 27, wherein IL-27 expression is assessed using the Tumor Proportion Score (TPS) and / or the Combined Positive Score (CPS).

29. 29. The method of claim 28, wherein the TPS or CPS is at least 10%, at least 20%, at least 30%, at least 50% or at least 60%.

30. The method of any one of claims 18 to 29, wherein IL-27 expression is detected by contacting the tumor sample with an antibody or antigen-binding portion thereof that specifically binds human IL-27.

31. The method of any one of claims 1 to 28, wherein the IL-27 inhibitor increases the expression of GBP5 and IRF1.

32. 30. The method of claim 29, wherein the IL-27 inhibitor increases the expression of GBP5 and IRF1 in NK cells and / or CD8+ T cells.

33. The method of any one of claims 1 to 32, wherein the IL-27 inhibitor reduces or blocks the interaction of IL-27 with WSX-1.

34. The method of any one of claims 1 to 33, wherein the IL-27 inhibitor comprises a polypeptide or a small molecule.

35. The method of any one of claims 1 to 34, wherein the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27 (an "anti-IL-27 antibody").

36. 36. The method of claim 35, wherein the anti-IL-27 antibody specifically binds to an epitope on human IL-27 comprising one or more amino acids of (i) amino acids 37 to 56 corresponding to SEQ ID NO:2 (IL-27p28), (ii) amino acids 142 to 164 corresponding to SEQ ID NO:2 (IL-27p28), or (iii) both (i) and (ii).

37. 37. The method of claim 35 or 36, wherein the epitope comprises one or more amino acids of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, or Glu164 of SEQ ID NO: 2 (IL-27p28).

38. 38. The method of claim 36 or 37, wherein the epitope comprises Asp146, Arg149, and / or Phe153 of SEQ ID NO: 2 (IL-27p28).

39. 39. The method of claim 38, wherein the epitope further comprises His150 and / or Leu156 of SEQ ID NO: 2 (IL-27p28).

40. 40. The method of claim 38 or 39, wherein the epitope further comprises Gln37, Leu38, Glu42, Leu142, and / or Glu164 of SEQ ID NO: 2 (IL-27p28).

41. The method of any one of claims 38 to 40, wherein the epitope further comprises Glu46, Val49, Ser50, and / or Leu162 of SEQ ID NO: 2 (IL-27p28).

42. 42. The method of any one of claims 36 to 41, wherein the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu142, Asp146, Arg149, His150, Phe153, Leu156, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28).

43. 42. The method of any one of claims 36-41, wherein the epitope further comprises one or more amino acids of Leu53, Lys56, Asp143, Leu147, Arg152, Ala157, Gly159, Phe160, or Asn161 of SEQ ID NO: 2 (IL-27p28).

44. 42. The method of any one of claims 36-41, wherein the epitope further comprises one or more amino acids of Leu53, Lys56, Asp143, Arg145, Leu147, Arg152, Ala157, Gly159, Phe160, Asn161, or Pro163 of SEQ ID NO:2 (IL-27p28).

45. 42. The method of any one of claims 36 to 41, wherein the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, and Glu164 of SEQ ID NO: 2 (IL-27p28).

46. 42. The method of any one of claims 36 to 41, wherein the epitope consists of or consists essentially of Gln37, Leu38, Glu42, Glu46, Val49, Ser50, Leu53, Lys56, Leu142, Asp143, Arg145, Asp146, Leu147, Arg149, His150, Arg152, Phe153, Leu156, Ala157, Gly159, Phe160, Asn161, Leu162, Pro163, and Glu164 of SEQ ID NO: 2 (IL-27p28).

47. 47. The method of any one of claims 35 to 46, wherein the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3, a light chain CDR1, a light chain CDR2, and a light chain CDR3, and wherein (i) the light chain CDR1 consists of N-XXXXXXLFSSNXKXYXX-C, the light chain CDR3 consists of N-XXXASAXXX-C, the heavy chain CDR2 consists of N-XXSSSSXSYXYXXXXXXXX-C, and the heavy chain CDR3 consists of N-XXXXGRTSYTATXHNXXXX-C, where X is any amino acid.

48. 48. The method of any one of claims 35 to 47, wherein the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, and the antibody or antigen-binding portion thereof comprises a heavy chain CDR3 comprising the sequence set forth in SEQ ID NO: 121 or 124.

49. 49. The method of any one of claims 35 to 48, wherein the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR2 comprising the sequence set forth in SEQ ID NO: 120 or 123.

50. 50. The method of any one of claims 35 to 49, wherein the IL-27 inhibitor comprises an antibody or antigen-binding portion thereof that specifically binds to human IL-27, and the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the sequence set forth in SEQ ID NO: 119 or 122.

51. 51. The method of any one of claims 35 to 50, wherein the antibody or antigen-binding portion thereof comprises a light chain CDR3 comprising the sequence set forth in SEQ ID NO: 129 or 132.

52. 52. The method of any one of claims 35 to 51, wherein the antibody or antigen-binding portion thereof comprises a light chain CDR2 comprising the sequence set forth in SEQ ID NO: 128 or 131.

53. 53. The method of any one of claims 35 to 52, wherein the antibody or antigen-binding portion thereof comprises a light chain CDR1 comprising the sequence set forth in SEQ ID NO: 127 or 130.

54. an antibody or antigen-binding portion thereof, (a) a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121; or (b) a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 122, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124; 54. The method of any one of claims 35 to 53, comprising:

55. an antibody or antigen-binding portion thereof, (a) a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; or (b) a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 132; 55. The method of any one of claims 35 to 54, comprising:

56. 56. The method of any one of claims 35 to 55, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 119, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 121, a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO:

129.

57. 57. The method of any one of claims 35 to 56, wherein the antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 122, a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 123, a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124, a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130, a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131, and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO:

132.

58. 58. The method of any one of claims 35-57, wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:

125.

59. 59. The method of any one of claims 35 to 58, wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:

125.

60. 60. The method of any one of claims 35-59, wherein the antibody or antigen-binding portion thereof comprises a light chain variable region comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:

133.

61. 61. The method of any one of claims 35 to 60, wherein the antibody or antigen-binding portion thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:

133.

62. 62. The method of any one of claims 35 to 61, wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 125 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:

133.

63. 63. The method of any one of claims 35 to 62, wherein the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:

135.

64. 64. The method of any one of claims 35 to 63, wherein the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:

139.

65. 65. The method of any one of claims 35 to 64, wherein the antibody or antigen-binding portion thereof comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:

137.

66. 66. The method of any one of claims 35 to 65, wherein the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 135 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

137.

67. 67. The method of any one of claims 35 to 66, wherein the antibody or antigen-binding portion thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 139 and a light chain comprising the amino acid sequence set forth in SEQ ID NO:

137.

68. 68. The method of any one of claims 1 to 67, wherein the cancer is selected from lung cancer (e.g., non-small cell lung cancer), sarcoma, testicular cancer, ovarian cancer, pancreatic cancer, breast cancer (e.g., triple-negative breast cancer), melanoma, head and neck cancer (e.g., head and neck squamous cell carcinoma), colorectal cancer, bladder cancer, endometrial cancer, prostate cancer, thyroid cancer, hepatocellular carcinoma (HCC), gastric cancer, brain tumor, lymphoma (e.g., DL-BCL), leukemia (e.g., AML), kidney cancer (e.g., renal cell carcinoma (RCC), e.g., clear cell RCC and / or non-clear cell RCC), and any combination thereof.

69. 69. The method of any one of claims 1 to 68, further comprising administering to the subject an additional therapeutic agent.

70. 70. The method of claim 69, wherein the additional therapeutic agent is administered before, after, or simultaneously with the antibody or antigen-binding portion thereof.

71. 71. The method of claim 69 or 70, wherein the additional therapeutic agent comprises chemotherapy, targeted anti-cancer therapy, oncolytic agent, cytotoxic agent, immune-based therapy, cytokine, surgery, radiation treatment, activator of costimulatory molecules, inhibitor of inhibitory molecules, vaccine, cellular immunotherapy, biologic agent, or a combination thereof.

72. 72. The method of any one of claims 69-71, wherein the additional therapeutic agent comprises a PD-1 antagonist, a PD-L1 inhibitor, a TIM-3 inhibitor, a LAG-3 inhibitor, a TIGIT inhibitor, a CD112R inhibitor, a TAM inhibitor, a STING agonist, a 4-1BB agonist, a multi-tyrosine kinase inhibitor (e.g., a VEGFR inhibitor), an anti-VEGF blocking antibody, a CTLA-4 antagonist, a HIF2 antagonist, a TGFb antagonist, an mTOR inhibitor, an adenosine pathway inhibitor (e.g., an anti-CD73 antibody, an anti-CD39 antibody, an anti-A2AR antibody, an anti-A2BR, or any combination thereof), an anti-CCR8 antibody, a cytokine-based regimen (e.g., IL-2 or IFN-a), a PARP inhibitor, or a combination thereof.

73. 73. The method of any one of claims 69-72, wherein the additional therapeutic agent comprises a PD-1 antagonist.

74. 74. The method of claim 73, wherein the PD-1 antagonist is selected from the group consisting of PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, and AMP-224.

75. 75. The method of claim 74, wherein the PD-L1 inhibitor is selected from the group consisting of FAZ053, atezolizumab, avelumab, durvalumab, and BMS-936559.

76. The additional therapeutic agent may be sunitinib (SUTENT®), cabozantinib (CABOMETYX®), axitinib (INLYTA®), lenvatinib (LENVIMA®), everolimus (AFINITOR®), bevacizumab (AVASTIN®), epacadostat, NKTR-214 (CD-122 biased agonist), tivozanib (FOTIVDA®), abexinostat, ipilimumab (YERVOY®), tremelimumab, pazopanib (VOTRIENT®), sorafe Niraparib (NEXAVAR®), temsirolimus (TORISEL®), ramucirumab (CYRAMZA®), niraparib, savolitinib, borolanib (X-82), regorafenib (STIVARGO®), donafenib (multikinase inhibitor), camrelizumab (SHR-1210), pexastimodin devasilepvec (JX-594), ramucirumab (CYRAMZA®), apatinib (YN968D1), encapsulated doxorubicin (THERMODOX®), tivantinib (ARQ197), ADI-PEG 20, binimetinib, apatinib mesylate, nintedanib, lirilumab, nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFIMZI®), cemiplimab-rwlc (LIBTAYO®), tislelizumab, and spartalizumab.

77. 73. The method of claim 72, wherein the additional therapeutic agent is a TIM-3 inhibitor.

78. 78. The method of claim 77, wherein the TIM-3 inhibitor is MGB453 or TSR-022.

79. 73. The method of claim 72, wherein the additional therapeutic agent is a LAG-3 inhibitor.

80. 80. The method of claim 79, wherein the LAG-3 inhibitor is selected from the group consisting of LAG525, BMS-986016, and TSR-033.

81. 73. The method of claim 72, wherein the additional therapeutic agent is a TIGIT inhibitor.

82. 73. The method of claim 72, wherein the additional therapeutic agent is a CD112R inhibitor.

83. 73. The method of claim 72, wherein the additional therapeutic agent is a TAM (Axl, Mer, Tyro) inhibitor.

84. 73. The method of claim 72, wherein the additional therapeutic agent is a 4-1BB agonist.

85. 73. The method of claim 72, wherein the additional therapeutic agent is a tyrosine kinase inhibitor (TKI).

86. (i) an antibody or antigen-binding portion thereof that specifically binds to human WSX-1; (ii) an IL-27 inhibitor; and (iii) Instructions for using (i) and (ii) in the method of any one of claims 1 to 85. Kit including:

87. (i) an antibody or antigen-binding portion thereof that specifically binds to human IL-27; (ii) an IL-27 inhibitor; and (iii) Instructions for using (i) and (ii) in the method of any one of claims 1 to 85. Kit including:

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